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• Correction Volume 14, Issue 8 pp 3720-3721

  Correction for: CD36 upregulates DEK transcription and promotes cell migration and invasion via GSK-3β/β-catenin-mediated epithelial-to-mesenchymal transition in gastric cancer

  Relevance score: 18.195171

  Jin Wang, Ti Wen, Zhi Li, Xiaofang Che, Libao Gong, Zihan Jiao, Xiujuan Qu, Yunpeng Liu

  Keywords: CD36, epithelial-to-mesenchymal transition, gastric cancer, DEK

  Published in Aging on April 29, 2022

• Research Paper Volume 13, Issue 5 pp 7499-7516

  BDKRB2 is a novel EMT-related biomarker and predicts poor survival in glioma

  Relevance score: 18.195171

  Ying Yang, Jin Wang, Fei Shi, Aijun Shan, Shihai Xu, Wen Lv

  Keywords: glioma, BDKRB2, epithelial-to-mesenchymal transition, EMT, prognosis

  Published in Aging on March 3, 2021

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  Bradykinin receptor B2 (BDKRB2) has been reported as an oncogene in several malignancies. In glioma, the role of BDKRB2 remains unknown. This study aimed at investigating its clinical significance and biological function in glioma at the transcriptional level. We selected 301 glioma patients with microarray data from CGGA database and 697 with RNAseq data from TCGA database. Transcriptome and clinical data of 998 samples were analyzed. Statistical analysis and figure generating were performed with R language. BDKRB2 expression showed a positive correlation with the WHO grade of glioma. BDKRB2 was increased in IDH wildtype and mesenchymal subtype of glioma. Gene ontology analysis demonstrated that BDKRB2 was profoundly associated with extracellular matrix organization in glioma. GSEA analysis revealed that BDKRB2 was particularly correlated with epithelial-to-mesenchymal transition (EMT). GSVA analysis showed that BDKRB2 was significantly paralleled with several EMT signaling pathways, including PI3K/AKT, hypoxia, and TGF-β. Moreover, BDKRB2 expression was significantly correlated with key biomarkers of EMT, especially with N-cadherin, snail, slug, vimentin, TWIST1, and TWIST2. Finally, higher BDKRB2 indicated significantly shorter survival for glioma patients. In conclusion, BDKRB2 was associated with more aggressive phenotypes of gliomas. Furthermore, BDKRB2 was involved in the EMT process and could serve as an independent prognosticator in glioma.

• Research Paper Volume 13, Issue 4 pp 4999-5019

  HOXB9 enhances the ability of lung cancer cells to penetrate the blood-brain barrier

  Relevance score: 13.273514

  HongShan Zheng, ChenLong Li, ZhenZhe Li, KaiBin Zhu, HongBo Bao, JinSheng Xiong, Peng Liang

  Keywords: brain metastasis, non-small cell lung cancer, blood-brain barrier, HOXB9, epithelial-to-mesenchymal transition

  Published in Aging on December 19, 2020

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  Even after multimodal therapy, the prognosis is dismal for patients with brain metastases from non-small cell lung cancer (NSCLC). Although the blood-brain barrier (BBB) limits tumor cell penetration into the brain parenchyma, some nevertheless colonize brain tissue through mechanisms that are not fully clear. Here we show that homeobox B9 (HOXB9), which is commonly overexpressed in NSCLC, promotes epithelial-to-mesenchymal transition (EMT) and tumor migration and invasion. Animal experiments showed that HOXB9 expression correlates positively with the brain metastatic potential of human NSCLC cells, while brain metastatic cells derived through in vivo selection showed greater HOXB9 expression than their cells of origin. Comparable results were obtained after immunohistochemical analysis of clinical primary NSCLC and matched brain metastasis samples obtained after surgery. Using an in vitro BBB model, knockdown and overexpression experiments showed that HOXB9-dependent expression of MMP9 in NSCLC cells leads to reduced expression of junctional proteins in cultured human vascular endothelial cells and enhanced transmigration of tumor cells. These data indicate that HOXB9 enables NSCLC cells to break away from the primary tumor by inducing EMT, and promotes brain metastasis by driving MMP9 production and degradation of intercellular adhesion proteins in endothelial cells comprising the BBB.

• Research Paper Volume 13, Issue 2 pp 1883-1897

  CD36 upregulates DEK transcription and promotes cell migration and invasion via GSK-3β/β-catenin-mediated epithelial-to-mesenchymal transition in gastric cancer

  Relevance score: 18.195171

  Jin Wang, Ti Wen, Zhi Li, Xiaofang Che, Libao Gong, Zihan Jiao, Xiujuan Qu, Yunpeng Liu

  Keywords: CD36, epithelial-to-mesenchymal transition, gastric cancer, DEK

  Published in Aging on November 21, 2020

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  Evidence indicates that the lipid scavenger receptor CD36 has pro-metastatic functions in several cancers. Although CD36 expression correlates with an unfavorable prognosis in gastric cancer (GC), its specific contribution to disease onset, progression, and/or metastasis remains unclear. Using bioinformatics analyses, we ascertained that CD36 expression was increased in metastatic GC specimens in The Cancer Genome Atlas and Gene Expression Omnibus databases and correlated with poor prognosis. In addition, higher CD36 expression was associated with lymph node metastasis (p < 0.05) and poor prognosis (p = 0.030) in 79 Chinese GC patients. Basal CD36 expression levels correlated positively with migration, invasion, and expression of epithelial-to-mesenchymal transition (EMT) markers in GC cell lines, a relationship confirmed by knockdown and overexpression experiments. Importantly, analysis of gene expression changes in CD36-knockdown GC cells led us to identify the chromatin-associated protein DEK as a c-Myc target that mediates activation of the GSK-3β/β-catenin signaling pathway to trigger EMT. These findings further our understanding of the mechanisms governing metastatic dissemination of GC cells and suggest the therapeutic potential of strategies targeting CD36.

• Research Paper Volume 12, Issue 14 pp 14141-14156

  Long noncoding RNA AC092171.4 promotes hepatocellular carcinoma progression by sponging microRNA-1271 and upregulating GRB2

  Relevance score: 17.377037

  Chengjun Sun, Shanzhou Huang, Yuchen Hou, Zhongqiu Li, Dongmei Xia, Lishan Zhang, Yixi Zhang, Yifeng Cai, Ziming Wang, Qi Zhou, Xiaoshun He, Linwei Wu

  Keywords: cancer, hepatocellular carcinoma, AC092171.4, epithelial-to-mesenchymal transition, survival

  Published in Aging on July 21, 2020

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  In this study, we investigated the mechanistic role of the long non-coding RNA (lncRNA) AC092171.4 in hepatocellular carcinoma (HCC). AC092171.4 was significantly upregulated in HCC tumor tissues compared to normal liver tissues. HCC patients with high AC092171.4 expression showed poorer overall survival (OS) and disease-free survival (DFS) than those with low AC092171.4 expression. In vitro cell proliferation, migration and invasiveness were all higher in AC092171.4-overexpressing HCC cells, but lower in AC092171.4-silenced HCC cells, than in controls. Balb/c nude mice injected with AC092171.4-silenced HCC cells had smaller xenograft tumors, which showed less growth and pulmonary metastasis than control tumors. Bioinformatics analyses and dual luciferase reporter assays confirmed that AC092171.4 binds directly to miR-1271, which targets the 3’UTR of GRB2 mRNA. AC092171.4 expression correlates negatively with miR1271 expression and correlates positively with GRB2 mRNA expression in HCC tissues from patients. HCC cells co-transfected with miR-1271 mimics and sh-AC092171.4 show less proliferation, migration, invasiveness, GRB2 protein, and epithelial to mesencyhmal transition (EMT) than sh-AC092171.4-transfected HCC cells. These findings demonstrate that AC092171.4 promotes growth and progression of HCC by sponging miR-1271 and upregulating GRB2. This makes AC092171.4 a potential prognostic indicator and therapeutic target for HCC patients.

  

  AC092171.4 is upregulated in HCC tissues. (A) Quantitative real-time PCR (qRT-PCR) analysis of AC092171.4 expression in HCC (n=369) and normal liver (n=50) tissues from the GEPIA database. As shown, AC092171.4 levels are higher in HCC tissues compared to normal liver tissues (^*p<0.05). (B) QRT-PCR analysis of AC092171.4 expression in 70 pairs of HCC and corresponding ANLTs. As shown, AC092171.4 levels are higher in HCC tissues compared to adjacent normal liver tissues (ANLTs; ^****p<0.0001). (C) QRT-PCR analysis showing AC092171.4 expression was significantly higher in the Huh7 and LM3 HCC cell lines than PLC/PRF/5 and Hep3B cells. (D) Chromogenic in situ hybridization (CISH) analysis of AC092171.4 expression in 95 pairs of HCC and ANLTs in a representative photograph. The results show that AC092171.4 expression is higher in HCC tissues compared to ANLTs (p<0.001). (E) Kaplan-Meier survival curve analyses of overall survival (OS) and disease-free survival (DFS) in HCC patients with low (n=42) and high (n=53) AC092171.4 expression. (F) Kaplan-Meier survival curve analyses of overall survival (OS) and disease-free survival (DFS) in high and low AC092171.4-expressing HCC patients of the GEPIA dataset (tumor=369, normal=50). * denotes p<0.05.

  
  
  

  

  AC092171.4 silencing decreases proliferation, migration and invasion in HCC cell lines. (A) QRT-PCR analysis shows AC092171.4 expression in the sh-NC and sh-AC092171.4 transfected Huh7 and LM3 cells. (B) CCK-8 assay results show proliferation status of the sh-NC and sh-AC092171.4 transfected Huh7 and LM3 cells. (C) EdU assay results show proliferation status of the sh-NC and sh-AC092171.4 transfected Huh7 cells. (D) Colony formation assay results show the total number of colonies in the sh-NC and sh-AC092171.4 transfected Huh7 cells. (E) EdU assay results show proliferation status of the sh-NC and sh-AC092171.4 transfected LM3 cells. (F) Colony formation assay results show the total number of colonies in the sh-NC and sh-AC092171.4 transfected LM3 cells. (G) Transwell migration assay results show AC092171.4 downregulation inhibited cell migration and invasion after transfection. (H) Transwell invasion assay results show the numbers of invasive sh-NC and sh-AC092171.4 transfected Huh7 and LM3 cells. (I) Representative western blot shows the expression of E-cadherin (epithelial marker) as well as N-cadherin and vimentin (mesenchymal markers) in the sh-NC and sh-AC092171.4 transfected Huh7 and LM3 cells. β-actin was used as loading control. * denotes p<0.05.

  
  
  

  

  AC092171.4 overexpression increases proliferation, invasion and migration in HCC cell lines. (A) QRT-PCR analysis shows AC092171.4 levels in the control and AC092171.4 overexpressing PLC/PRF/5 and Hep3B cells. (B) CCK-8 assay results show cell proliferation status of control and AC092171.4 overexpressing PLC/PRF/5 and Hep3B cells. (C) EdU assay results show proliferation status of control and AC092171.4 overexpressing PLC/PRF/5 cells. (D) EdU assay results show proliferation status of control and AC092171.4 overexpressing Hep3B cells. (E) Colony formation assay results show the total number of colonies in control and AC092171.4 overexpressing PLC/PRF/5 cells. (F) Colony formation assay results show the total number of colonies in control and AC092171.4 overexpressing Hep3B cells. (G and H) Transwell migration assay results show the total numbers of migrating control and AC092171.4 overexpressing PLC/PRF/5 and Hep3B cells. (I and J) Transwell invasion assay results show the total numbers of invasive control and AC092171.4 overexpressing PLC/PRF/5 and Hep3B cells. (K) Representative western blot shows the expression of E-cadherin (epithelial marker) as well as N-cadherin and vimentin (mesenchymal markers) in control and AC092171.4 overexpressing PLC/PRF/5 and Hep3B cells. * denotes p<0.05.

  
  
  

  

  AC092171.4 knockdown represses in vivo xenograft HCC tumor growth and pulmonary metastases. (A) Balb/c nude mice were subcutaneously injected with sh-NC or sh-AC092171.4 transfected Huh7 cells and tumor from respective groups were shown (n=5). (B, C) Xenograft tumor volume and weights in nude mice subcutaneously injected with sh-NC or sh-AC092171.4 transfected Huh7 cells. (D, E) Immunohistochemical analysis shows percentage of Ki-67-positive stained cells in the sections of xenograft tumors from control and AC092171.4 knockdown nude mice. (F, G) Total numbers of pulmonary metastatic nodules in nude mice injected with control and AC092171.4 knockdown Huh-7 cells through the tail vein. * denotes p<0.05.

  
  
  

  

  AC092171.4 regulates GRB2 protein expression by competitively binding miR-1271. (A) QRT-PCR analysis shows miR-1271 levels in sh-NC and sh-AC092171.4-transfected HCC cells. (B) QRT-PCR analysis shows miR-1271 levels in control and AC092171.4 overexpressing HCC cells. (C) Dual luciferase reporter assay results show relative firefly luciferase activity in HCC cells transfected with wild-type or mutant AC092171.4 and miR-1271 mimics. (D) Pearson’s correlation analysis shows the association between AC092171.4 and miR-1271 levels in 45 HCC tissue samples from 70 pair HCC specimens. (E) Dual luciferase reporter assay results show relative firefly luciferase activity in HCC cells transfected with WT or mutant 3’UTR of GRB2 and miR-1271 mimics. (F) Western blot results show GRB2 protein levels in HCC cells transfected with miR-1271 mimics or miR-1271 inhibitors. (G) Pearson’s correlation analysis shows the relationship between AC092171.4 and GRB2 mRNA levels in HCC tissues from the TCGA datasets in GEPIA website. (H) Pearson’s correlation analysis shows the relationship between AC092171.4 and GRB2 mRNA levels in HCC tissues from the TCGA datasets. (I) Western blot analysis shows GRB2 protein expression in HCC cells transfected with AC092171.4-shRNA plus miR-1271 inhibitor or AC092171.4-shRNA alone. * denotes p<0.05.

  
  
  

  

  AC092171.4 promotes proliferation, migration, and invasiveness of HCC cells by suppressing miR-1271-dependent downregulation of GRB2 protein translation. (A) CCK-8 assay results show proliferation of Huh-7 and LM3 cells transfected with sh-NC, sh-AC092171.4, sh-NC plus miR-1271 inhibitor, or sh-AC092171.4 plus miR-1271 inhibitor. (B) Colony formation assays show the numbers of colonies formed by Huh-7 cells transfected with sh-NC, sh-AC092171.4, sh-NC plus miR-1271 inhibitor, or sh-AC092171.4 plus miR-1271 inhibitor. (C) Transwell assay results show the total numbers of migratory and invasive Huh-7 cells transfected with sh-NC, sh-AC092171.4, sh-NC plus miR-1271 inhibitor, or sh-AC092171.4 plus miR-1271 inhibitor. (D) Colony formation assays show the numbers of colonies formed by LM3 cells transfected with sh-NC, sh-AC092171.4, sh-NC plus miR-1271 inhibitor, or sh-AC092171.4 plus miR-1271 inhibitor. (E) Transwell assay results show the total numbers of migratory and invasive LM3 cells transfected with sh-NC, sh-AC092171.4, sh-NC plus miR-1271 inhibitor, or sh-AC092171.4 plus miR-1271 inhibitor. (F) Western blot analysis shows relative levels of E-cadherin, N-cadherin and vimentin levels in HCC cells transfected with shRNA-AC092171.4 alone or shRNA-AC092171.4 plus miR-1271 inhibitor. (G) CCK-8 assay results show the proliferation status in AC092171.4-silenced and sh-AC092171.4-silenced plus GRB2 overexpressing Huh7and LM3 cells. (H, I) Transwell assay results show the migration status of AC092171.4-silenced and sh-AC092171.4-silenced plus GRB2 overexpressing Huh7and LM3 cells. * denotes p<0.05.

  
  
  

• Research Paper Volume 12, Issue 3 pp 2333-2346

  Long non-coding RNA NNT-AS1 promotes cholangiocarcinoma cells proliferation and epithelial-to-mesenchymal transition through down-regulating miR-203

  Relevance score: 14.187396

  Yulei Gu, Zhiqiang Zhu, Hui Pei, Dong Xu, Yumin Jiang, Luanluan Zhang, Lili Xiao

  Keywords: long non-coding RNA NNT-AS1, cholangiocarcinoma, proliferation, epithelial-to-mesenchymal transition, miR-203

  Published in Aging on February 5, 2020

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  Background: Cholangiocarcinoma (CCA) is a serious malignant tumor. Long non-coding RNA NNT-AS1 (NNT-AS1) takes crucial roles in several tumors. So, we planned to research the roles and underlying mechanism of NNT-AS1 in CCA.

  Results: NNT-AS1 overexpression was appeared in CCA tissues and cell lines. Proliferation was promoted by NNT-AS1 overexpression in CCLP1 and TFK1 cells. Besides, NNT-AS1 overexpression reduced E-cadherin level and raised levels of N-cadherin, vimentin, Snail and Slug. However, the opposite trend was occurred by NNT-AS1 knockdown. Further, NNT-AS1 overexpression promoted phosphatidylinositol 3 kinase (PI3K)/AKT and extracellular signal-regulated kinase (ERK)1/2 pathways. MiR-203 was sponged by NNT-AS1 and miR-203 mimic reversed the above promoting effects of NNT-AS1. Additionally, insulin-like growth factor type 1 receptor (IGF1R) and zinc finger E-box binding homeobox 1 (ZEB1) were two potential targets of miR-203.

  Conclusion: NNT-AS1 promoted proliferation, EMT and PI3K/AKT and ERK1/2 pathways in CCLP1 and TFK1 cells through down-regulating miR-203.

  Methods: CCLP1 and TFK1 cells were co-transfected with pcDNA-NNT-AS1 and miR-203 mimic. Bromodeoxyuridine (BrdU), flow cytometry, quantitative reverse transcription polymerase chain reaction (qRT-PCR) and western blot were employed to detect roles and mechanism of NNT-AS1. Interaction between NNT-AS1 and miR-203 or miR-203 and target genes was examined through luciferase activity experiment.

  

  NNT-AS1 overexpression was occurred in CCA. NNT-AS1 levels in CCA tissues (A) and cell lines (B) were measured through qRT-PCR. * P < 0.05 and ** P < 0.01 contrasted with indicated group.

  
  
  

  

  Effects of NNT-AS1 on the growth of CCLP1 and TFK1 cells, which were transfected with pcDNA-NNT-AS1 and si-NNT-AS1. (A) NNT-AS1 level was examined via qRT-PCR. (B) Proliferation was examined via BrdU. Expression of cyclinD1 was examined via western blot (C) and analyzed quantitatively (D) in both cells. (E) Apoptosis was examined via flow cytometry. Expression of cleaved-caspase-3 was examined via western blot (F) and analyzed quantitatively (G) in both cells. * P < 0.05, ** P < 0.01 and *** P < 0.001 contrasted with indicated set.

  
  
  

  

  Effect of NNT-AS1 on EMT in CCLP1 and TFK1 cells, which were transfected with pcDNA-NNT-AS1 and si-NNT-AS1. Expression of EMT relative factors was examined via western blot (A, C) and analyzed quantitatively (B, D) in CCLP1 and TFK1 cells. * P < 0.05 and ** P < 0.01 contrasted with indicated set.

  
  
  

  

  The interaction between NNT-AS1 and miR-203 or miR-203 and IGF1R/ZEB1 was examined. (A) The target matching sequence of NNT-AS1 and miR-203. (B) Luciferase activity was measured after co-transfection with NNT-AS1-wt or NNT-AS1-mut and miR-203 mimic or NC mimic. (C) Level of miR-203 was measured via qRT-PCR when cells were transfected with pcDNA-NNT-AS1 and si-NNT-AS1. (D) The target matching sequence of miR-203 and IGF1R/ZEB1. (E–F) Luciferase activities were measured after co-transfection with IGF1R/ZEB1-wt or IGF1R/ZEB1-mut and miR-203 mimic or NC mimic. (G–H) The mRNA and protein levels of IGF1R and ZEB1 were tested through qRT-PCR and western blot after transfection with miR-203 mimic or miR-203 inhibitor and the relative control. * P < 0.05 and ** P < 0.01 contrasted with indicated group.

  
  
  

  

  Mechanism of NNT-AS1 on proliferation and EMT in CCLP1 and TFK1 cells after co-transfection with pcDNA-NNT-AS1 and miR-203 mimic. (A) Standard of miR-203 was examined via qRT-PCR by transfection with miR-203 mimic. (B) Proliferation was examined via BrdU. Expression of cyclinD1 was examined via western blot (C) and analyzed quantitatively (D) in both cells. Expression of EMT related factors was inspected via western blot (E, G) and analyzed quantitatively (F, H) in CCLP1 and TFK1 cells. * P < 0.05 and ** P < 0.01 contrasted with indicated set.

  
  
  

  

  Mechanism of NNT-AS1 on PI3K/AKT and ERK1/2 passageways in CCLP1 and TFK1 cells, which were co-transfected with pcDNA-NNT-AS1 and miR-203 mimic. Expression of PI3K and AKT was measured via western blot (A, C) and analyzed quantitatively (B, D) in both cells. Expression of ERK1/2 was examined via western blot (E, G) and analyzed quantitatively (F, H) in both cells. * P < 0.05, ** P < 0.01 and *** P < 0.001 contrasted with indicated set.

  
  
  

  

  NNT-AS1 regulated other miRNAs in CCA. The expression of miR-363, miR-129-5p and miR-142-3p was examined via qRT-PCR in CCLP1 (A) and TFK1 (B) cells. * P < 0.05 and ** P < 0.01 contrasted with indicated set.

  
  
  

• Research Paper Volume 12, Issue 1 pp 866-883

  Prognostic value of epithelial-mesenchymal transition markers in clear cell renal cell carcinoma

  Relevance score: 13.764213

  Hua Xu, Wen-Hao Xu, Fei Ren, Jun Wang, Hong-Kai Wang, Da-Long Cao, Guo-Hai Shi, Yuan-Yuan Qu, Hai-Liang Zhang, Ding-Wei Ye

  Keywords: clear cell renal cell carcinoma, epithelial-to-mesenchymal transition, prognosis, tumor microenvironment, predictive model

  Published in Aging on January 8, 2020

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  Epithelial-to-mesenchymal transition (EMT) is important in tumor invasiveness and metastasis. We aimed to determine prognostic value of six key EMT markers (CDH1, CDH2, SNAI1, SNAI2, VIM, TWIST1) in clear cell renal cell carcinoma (ccRCC). A total of 533 ccRCC patients with RNASeq data from The Cancer Genome Atlas (TCGA) cohort were included for analysis. Gene expression of these EMT markers was compared between tumor and normal tissues based on Oncomine database and TCGA cohort. Their correlations with progression-free survival (PFS) and overall survival (OS) were also examined in both TCGA cohort and FUSCC (Fudan University Shanghai Cancer Center) cohort. Cox proportional hazards regression model and Kaplan-Meier plot were used to assess the relative factors. Functional enrichment analyses were utilized to describe biologic function annotations and significantly involved hallmarks pathways of each gene. We found that Epithelial marker, CDH1 expression was lower, while mesenchymal markers (CDH2, SNAI1, VIM, TWIST1) expression was higher in ccRCC primary tumors. In the TCGA cohort, we found that patients with higher expression of VIM, TWIST1 or lower expression of CDH1 had worse prognosis. Further, in the FUSCC cohort, we confirmed the predictive ability of mesenchymal markers and epithelial marker expression in PFS and OS of ccRCC patients. After generating Cox regression models, EMT markers (CDH1, SNAI1, VIM, and TWIST1) were independent prognostic factors of both PFS and OS in ccRCC patients. Our preliminary EMT prediction model can facilitate further screening of EMT biomarkers and cast a better understanding of EMT gene function in ccRCC.

  

  Analysis of the six EMT related genes in Oncomine database and TCGA database. (A) The Oncomine database was queried for the expression of CDH1, CDH2, SNAI1, SNAI2, VIM, and TWIST1 in the available datasets based on the following criteria: 1) “Cancer Type”; 2) “Gene: CDH1, CDH2, SNAI1, SNAI2, VIM, or TWIST1”; 3) “Data Type: mRNA”; 4) “Analysis Type: Cancer vs Normal Analysis”, and 5) Threshold Setting Condition (p<0.001, fold change >2, gene rank = top 10%). The 'red cells' represents gene overexpression and the 'blue cells' represent gene underexpression. The color intensity equals the percentile, i.e. Top 1%, 5%, or 10% significantly over- or underexpressed (see the legend below the grid). We found that CDH1 and SNAI2 was underexpressed in the kidney cancer vs normal datasets, while VIM was highly overexpressed. (B–G) Differential mRNA expression of six EMT related genes in clear cell renal cell carcinoma (ccRCC) tumor samples and adjacent normal tissues from TCGA. Epithelial marker CDH1 mRNA expression was significantly lower in tumor samples compared with adjacent normal tissues (B); Most mesenchymal markers (CDH2, SNAI1, VIM, and TWIST1) mRNA expression was elevated in tumor samples compared with adjacent normal tissues (C–G).

  
  
  

  

  Kaplan Meier survival plot of ccRCC patients in TCGA database according to high and low mRNA expression of six EMT related genes.CDH1 mRNA expression was associated with both worse progression-free survival (p=0.015) and worse overall survival (p=0.003) of ccRCC patients (A, G); CDH2 mRNA expression was not an indicator of either progression-free survival (p=0.593) or overall survival (p=0.075) of ccRCC patients (B, H); Higher SNAI1 mRNA expression was moderately associated with both worse progression-free survival (p=0.054) and worse overall survival (p=0.010) of ccRCC patients (C, I); SNAI2 mRNA expression was not an indicator of either progression-free survival (p=0.105) or overall survival (p=0.242) of ccRCC patients (D, J); Higher VIM mRNA expression was associated with both worse progression-free survival (p<0.001) and worse overall survival (p=0.005) of ccRCC patients (E, K); Higher TWIST1 mRNA expression was associated with both worse progression-free survival (p<0.001) and worse overall survival (p<0.001) of ccRCC patients (F, L).

  
  
  

  

  Kaplan Meier survival plot of ccRCC patients in FUSCC cohort according to high and low mRNA expression of six EMT related genes. Lower CDH1 mRNA expression was associated with both worse progression-free survival (p=0.016) and worse overall survival (p<0.001) of ccRCC patients (A, G); CDH2 mRNA expression was not an indicator of either progression-free survival (p=0.288) or overall survival (p=0.202) of ccRCC patients (B, H); Higher SNAI1 mRNA expression was associated with both worse progression-free survival (p<0.001) and worse overall survival (p<0.001) of ccRCC patients (C, I); Higher SNAI2 mRNA expression was associated with both worse progression-free survival (p=0.005) and worse overall survival (p<0.001) of ccRCC patients (D, J); Higher VIM mRNA expression was associated with both worse progression-free survival (p<0.001) and worse overall survival (p<0.001) of ccRCC patients (E, K); Higher TWIST1 mRNA expression was associated with both worse progression-free survival (p<0.001) and worse overall survival (p<0.001) of ccRCC patients (F, L).

  
  
  

  

  Construction and internal validation of integrated prognostic and diagnostic model. All significant clinicopathologic parameters and gene expression profiles was integrated in the Cox regression models, which indicated this formula: = -0.708×CDH1 expression (ref. Low) + 1.360×SNAI1 expression (ref. Low) + 1.905×VIM expression (ref. Low) + 2.179×TWIST1 expression (ref. Low) + 1.274×T stage (ref. T1-T2) + 1.919×M stage (ref. M0) + 2.021×AJCC stage (ref. I-II) + 2.013×ISUP grade (ref. 1-2) for PFS (A), and another formula: = -0.564×CDH1 expression (ref. Low) + 1.532×SNAI1 expression (ref. Low) + 1.804×VIM expression (ref. Low) + 1.714×TWIST1 expression (ref. Low) + 1.226×T stage (ref. T1-T2) + 1.778×M stage (ref. M0) + 2.515×AJCC stage (ref. I-II) + 1.954×ISUP grade (ref. 1-2) for OS (B). The Kaplan–Meier method was used to determine the significant survival outcomes (PFS: p<0.0001; OS: p<0.0001). ROC curves were generated to validate the ability of the logistic model to predict prognosis. The AUC index for the integrated model were 0.886 for PFS (p<0.001) (C) and 0.814 for OS (p<0.001) (D). (E, F) External validation of integrated model using TCGA cohorts. ROC curves were constructed to perform external validation using clinicopathological parameters and mRNA expression profiles from TCGA cohort. The AUC index for the integrated model were 0.720 for PFS (p<0.001) (E) and 0.684 for OS (p<0.001) (F).

  
  
  

  

  CDH1, CDH2, SNAI1, SNAI2 and TWIST1 significantly involved in stromal process of ccRCC tumor environment. EMT makers, including CDH1 (A), CDH2 (B), SNAI1 (C), SNAI2 (D), TWIST1 (F), but not VIM (E), showed significant correlation with stromal process in ccRCC environments (p<0.001). In addition, CDH1 showed negative association with stromal score (r²=-0.191), while stromal score positively correlated CDH2 (r²=0.337), SNAI1 (r²=0.199), SNAI2 (r²=0.201) and TWIST1 (r²=0.305) mRNA expression in ccRCC patients from TCGA cohort.

  
  
  

  

  Module analysis and functional annotations of the six EMT related gene in silico. Protein-protein interaction (PPI), activation and indirect relation were predicted and displayed in association with sig EMT related genes (A). PPI network derived from active interaction sources was detailed illustrated with required interaction score equal 0.400 (B). GO and KEGG functional annotations analysis of CDH1, CDH2, SNAI1, SNAI2, VIM, TWIST1 was enriched in hemophilic cell adhesion and cell-cell adhesion of biologic process, adherens junction, anchoring junction and cell junction of cellular component, RPTP-like protein binding, phosphatase binding, protein phosphatase binding and enzyme binding of molecular function. Participating upstream or downstream signaling pathways enrichment include adherens junction, cell adhesion molecules (CAMs) of KEGG pathways (C). Hierarchical partitioning using transcriptional expression profiles of CDH1, CDH2, SNAI1, SNAI2, VIM, TWIST1 from FUSCC cohort (D). Hierarchical partitioning using transcriptional expression profiles of six hub genes from TCGA cohort was performed in the heat map (E).

  
  
  

• Research Paper Volume 10, Issue 12 pp 3662-3682

  Long noncoding RNA B3GALT5-AS1 suppresses colon cancer liver metastasis via repressing microRNA-203

  Relevance score: 15.311605

  Liang Wang, Zhewei Wei, Kaiming Wu, Weigang Dai, Changhua Zhang, Jianjun Peng, Yulong He

  Keywords: long noncoding RNA, colon cancer, liver metastasis, microRNA, epithelial-to-mesenchymal transition

  Published in Aging on December 10, 2018

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  Long noncoding RNAs (lncRNAs) are implicated in various cancers, including colon cancer. Liver metastasis is the main cause of colon cancer-related death. However, the roles of lncRNAs in colon cancer liver metastasis are still largely unclear. In this study, we identified a novel lncRNA B3GALT5-AS1, which is reduced in colon cancer tissues and further reduced in colon cancer liver metastasis tissues. Reduced expression of B3GALT5-AS1 is associated with liver metastasis and poor outcome of colon cancer patients. Gain-of-function and loss-of-function assays revealed that B3GALT5-AS1 inhibited proliferation but promoted migration and invasion of colon cancer cells. Further investigation revealed that B3GALT5-AS1 directly bound to the promoter of miRNA-203, repressed miR-203 expression, upregulated miR-203 targets ZEB2 and SNAI2, and induced epithelial-to-mesenchymal transition (EMT). In vivo study revealed that B3GALT5-AS1 suppressed colon cancer liver metastasis via its binding on miR-203 promoter and the repression of miR-203. miR-203 is increased and epithelial phenotype is preferred in colon cancer liver metastasis tissues. Collectively, our data revealed the suppressive roles of B3GALT5-AS1/miR-203/EMT regulation axis in colon cancer liver metastasis. Our data suggested that the activating B3GALT5-AS1/miR-203/EMT axis may be potential therapeutic strategy for colon cancer liver metastasis.

  

  The expression pattern of B3GALT5-AS1 in colon cancer and its association with prognosis. (A) The expression intensity of B3GALT5-AS1 in 18 pairs of normal colonic epithelium, primary colorectal cancers, and metastasized cancers in liver from GSE50760. (B) The expression of B3GALT5-AS1 in 64 pairs of primary colon cancer tissues and adjacent colonic epithelium tissues was detected using qRT-PCR. P < 0.0001, Wilcoxon signed-rank test. (C) The expression of B3GALT5-AS1 in 15 colon cancer tissues with metastasis and 49 colon cancer tissues without metastasis. P < 0.0001, Mann-Whitney test. (D) The expression of B3GALT5-AS1 in 15 pairs of primary colon cancer tissues and corresponding liver metastasis tissues was measured using qRT-PCR. P < 0.0001, Wilcoxon signed-rank test. (E) Kaplan-Meier survival analysis of the correlation between B3GALT5-AS1 expression level and overall survival of 64 colon cancer patients. The median expression level of B3GALT5-AS1 was used as cut-off. P = 0.0096, Log-rank test. (F) The expression of B3GALT5-AS1 in normal colonic epithelial cell line NCM460 and colon cancer cell lines HCT116, HT-29, LoVo, SW480 and SW620 was measured using qRT-PCR. Results are displayed as mean ± s.d. of three independent experiments. **P < 0.01, ***P < 0.001, Student’s t-test.

  
  
  

  

  B3GALT5-AS1 suppressed colon cancer cell proliferation. (A) The expression of B3GALT5-AS1 in B3GALT5-AS1 stably overexpressed and control HCT116 cells was detected using qRT-PCR. (B) The expression of B3GALT5-AS1 in B3GALT5-AS1 stably depleted and control SW620 cells was detected using qRT-PCR. (C) Cell viability of B3GALT5-AS1 stably overexpressed and control HCT116 cells was detected using Glo cell viability assay. (D) Cell viability of B3GALT5-AS1 stably depleted and control SW620 cells was detected using Glo cell viability assay. (E) Cell proliferation of B3GALT5-AS1 stably overexpressed and control HCT116 cells was detected using EdU incorporation assay. The red color indicts EdU-positive cells. Scale bars = 100 μm. (F) Cell proliferation of B3GALT5-AS1 stably depleted and control SW620 cells was detected using EdU incorporation assay. The red color indicts EdU-positive cells. Scale bars = 100 μm. Results are displayed as mean ± s.d. of three independent experiments. **P < 0.01, ***P < 0.001, Student’s t-test.

  
  
  

  

  B3GALT5-AS1 promoted migration, invasion, and EMT of colon cancer cells. (A) Cell migration of B3GALT5-AS1 stably overexpressed and control HCT116 cells was detected using transwell migration assay. Scale bars = 100 μm. (B) Cell migration of B3GALT5-AS1 stably depleted and control SW620 cells was detected using transwell migration assay. Scale bars = 100 μm. (C) Cell invasion of B3GALT5-AS1 stably overexpressed and control HCT116 cells was detected using transwell invasion assay. Scale bars = 100 μm. (D) Cell invasion of B3GALT5-AS1 stably depleted and control SW620 cells was detected using transwell invasion assay. Scale bars = 100 μm. (E) E-cadherin and N-cadherin mRNA levels in B3GALT5-AS1 stably overexpressed and control HCT116 cells were detected using qRT-PCR. (F) E-cadherin and N-cadherin protein levels in B3GALT5-AS1 stably overexpressed and control HCT116 cells were detected using western blot. (G) E-cadherin and N-cadherin mRNA levels in B3GALT5-AS1 stably depleted and control SW620 cells were detected using qRT-PCR. (H) E-cadherin and N-cadherin protein levels in B3GALT5-AS1 stably depleted and control SW620 cells were detected using western blot. Results are displayed as mean ± s.d. of three independent experiments. *P < 0.05, **P < 0.01, Student’s t-test.

  
  
  

  

  B3GALT5-AS1 bound to the promoter of miR-203 and repressed the expression of miR-203. (A) The subcellular distribution of B3GALT5-AS1 in the cytoplasmic and nuclear fractions of HCT116 cells was evaluated using cytoplasmic and nuclear RNA isolation followed by qRT-PCR.β-actin and U6 were used as cytoplasmic and nuclear controls, respectively. (B) Schematic outline of the predicted interaction sites between B3GALT5-AS1 and the promoter of miR-203. (C) The expression of miR-203 in B3GALT5-AS1 stably overexpressed and control HCT116 cells was detected using qRT-PCR. (D) The expression of miR-203 in B3GALT5-AS1 stably depleted and control SW620 cells was detected using qRT-PCR. (E) ChIRP assays in HCT116 cells were carried out with anti-sense probe sets specific for B3GALT5-AS1 or LacZ (negative control). The enriched DNA was measured using qRT-PCR with specific primers against miR-203 promoter. (F) Schematic outline of the constructed different depletion transcripts of B3GALT5-AS1. (G) After transient transfections of the different B3GALT5-AS1 expressing plasmids into HCT116 cells, miR-203 expression was measured using qRT-PCR. (H) After transient co-transfection of the firefly luciferase reporter containing the promoter of miR-203, renilla luciferase expression plasmid pRL-TK, and the different B3GALT5-AS1 expression plasmids into HCT116 cells, luciferase activities were detected using dual luciferase reporter assays. Results are displayed as the relative ratio of firefly luciferase activity to renilla luciferase activity. (I) After transient co-transfection of the firefly luciferase reporter containing the promoter of miR-203 and pRL-TK into B3GALT5-AS1 stably depleted and control SW620 cells, luciferase activities were measured by dual luciferase reporter assays. Results are shown as the relative ratio of firefly luciferase activity to renilla luciferase activity. (J) After transient transfections of the different B3GALT5-AS1 expressing plasmids into HCT116 cells, the expression of ZEB2 and SNAI2 was detected using qRT-PCR and western blot. (K) The expression of ZEB2 and SNAI2 in B3GALT5-AS1 stably depleted and control SW620 cells was detected using qRT-PCR and western blot. Data are displayed as mean ± s.d. of three independent experiments. **P < 0.01, ***P < 0.001, NS, not significant, Student’s t-test

  
  
  

  

  miR-203 expression pattern in colon cancer. (A) miR-203expression in 64 pairs of primary colon cancer tissues and adjacent colonic epithelium tissues was measured by qRT-PCR. P < 0.0001, Wilcoxon signed-rank test. (B) The correlation between B3GALT5-AS1 and miR-203 expression level in colon cancer tissues. n = 64, r = -0.7625, P < 0.0001, Pearson’s correlation analysis. (C) The expression of miR-203 in 15 pairs of primary colon cancer tissues and corresponding liver metastasis tissues was measured using qRT-PCR. P < 0.0001, Wilcoxon signed-rank test. (D) The expression of ZEB2 in 15 pairs of primary colon cancer tissues and corresponding liver metastasis tissues was measured using qRT-PCR. P = 0.0006, Wilcoxon signed-rank test. (E) The expression of SNAI2 in 15 pairs of primary colon cancer tissues and corresponding liver metastasis tissues was measured using qRT-PCR. P = 0.0034, Wilcoxon signed-rank test. (F) The expression of E-cadherin in 15 pairs of primary colon cancer tissues and corresponding liver metastasis tissues was measured using qRT-PCR. P < 0.0001, Wilcoxon signed-rank test. (G) The expression of N-cadherin in 15 pairs of primary colon cancer tissues and corresponding liver metastasis tissues was measured using qRT-PCR. P = 0.0002, Wilcoxon signed-rank test.

  
  
  

  

  B3GALT5-AS1 inhibited colon cancer liver metastasis. (A) B3GALT5-AS1 expression in different B3GALT5-AS1 stably overexpressed HCT116 cells clones was measured using qRT-PCR. Data are displayed as mean ± s.d. of three independent experiments. ***P < 0.001, Student’s t-test. (B) Indicated B3GALT5-AS1 stably overexpressed HCT116 cells were intra-splenic injected to establish liver metastasis. The amount of liver metastatic foci was assessed at the 42^th day after intra-splenic injection using HE staining. Scale bars = 1000 μm. (C) The expression of B3GALT5-AS1 and miR-203 in liver metastatic foci formed by these indicated B3GALT5-AS1 stably overexpressed HCT116 cells was detected using qRT-PCR. (D) Immunohistochemical staining of Ki67 in liver metastatic foci formed by these indicated B3GALT5-AS1 stably overexpressed HCT116 cells. Scale bars = 50 μm. (E) The expression of ZEB2 and SNAI2 in liver metastatic foci formed by these indicated B3GALT5-AS1 stably overexpressed HCT116 cells was measured using qRT-PCR. (F) The expression of E-cadherin and N-cadherin in liver metastatic foci formed by these indicated B3GALT5-AS1 stably overexpressed HCT116 cells was detected using qRT-PCR. For B-F, data are displayed as mean ± s.d. of six mice in each group. **P < 0.01, NS, not significant, Mann-Whitney test.

  
  
  

  

  Depletion of B3GALT5-AS1 promoted colon cancer liver metastasis in a miR-203-dependent manner. (A) miR-203 expression in B3GALT5-AS1 and miR-203 concurrently depleted and control SW620 cells was measured using qRT-PCR. Data are displayed as mean ± s.d. of three independent experiments. **P < 0.01, Student’s t-test. (B) B3GALT5-AS1 and miR-203 concurrently depleted and control SW620 cells were intra-splenic injected to establish liver metastasis. The amount of liver metastatic foci was detected at the 42^th day after intra-splenic injection using HE staining. Scale bars = 1000 μm. (C) The expression of B3GALT5-AS1 and miR-203 in liver metastatic foci formed by B3GALT5-AS1 and miR-203 concurrently depleted and control SW620 cells was detected using qRT-PCR. (D) Immunohistochemical staining of Ki67 in liver metastatic foci formed by B3GALT5-AS1 and miR-203 concurrently depleted and control SW620 cells. Scale bars = 50 μm. (E) The expression of ZEB2 and SNAI2 in liver metastatic foci formed by B3GALT5-AS1 and miR-203 concurrently depleted and control SW620 cells was measured using qRT-PCR. (F) The expression of E-cadherin and N-cadherin in liver metastatic foci formed by B3GALT5-AS1 and miR-203 concurrently depleted and control SW620 cells was detected using qRT-PCR. For B-F, data are displayed as mean ± s.d. of six mice in each group. **P < 0.01, Mann-Whitney test.

  
  
  

• Research Paper Volume 9, Issue 2 pp 524-546

  Compound effects of aging and experimental FSGS on glomerular epithelial cells

  Relevance score: 14.292585

  Remington R.S. Schneider, Diana G. Eng, J. Nathan Kutz, Mariya T. Sweetwyne, Jeffrey W. Pippin, Stuart J. Shankland

  Keywords: kidney disease, glomerulus, parietal epithelial cell, podocyte, epithelial to mesenchymal transition, Collagen IV

  Published in Aging on February 17, 2017

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  Advanced age portends a poorer prognosis in FSGS. To understand the impact of age on glomerular podocytes and parietal epithelial cells (PECs), experimental FSGS was induced in 3m-old mice (20-year old human age) and 27m-old mice (78-year old human age) by abruptly depleting podocytes with a cytopathic anti-podocyte antibody. Despite similar binding of the disease-inducing antibody, podocyte density was lower in aged FSGS mice compared to young FSGS mice. Activated PEC density was higher in aged versus young FSGS mice, as was the percentage of total activated PECs. Additionally, the percentage of glomeruli containing PECs with evidence of phosphorylated ERK and EMT was higher in aged FSGS mice. Extracellular matrix, measured by collagen IV and silver staining, was higher in aged FSGS mice along Bowman’s capsule. However, collagen IV accumulation in the glomerular tufts alone and in glomeruli with both tuft and Bowman’s capsule accumulation were similar in young FSGS and aged FSGS mice. Thus, the major difference in collagen IV staining in FSGS was along Bowman’s capsule in aged mice. The significant differences in podocytes, PECs and extracellular matrix accumulation between young mice and old mice with FSGS might explain the differences in outcomes in FSGS based on age.

  

  Albuminuria was higher in aged mice at baseline and in FSGS. (A-D) Sheep IgG staining. Sheep IgG staining confirmed the equal distribution of anti-glomerular antibody in the glomerular tuft in both young (Y) and aged (A) mice. Images were taken at 100x. (A'-D') 400x close-up images of glomeruli from (A-D). Sheep IgG deposition (purple color) was not seen in baseline mice as expected (A,B,A',B'). Animals given FSGS with the anti-glomerular antibody showed sheep IgG deposition in the glomerular tufts (C, D, C’, D'). (E) Albumin to creatinine ratios (ACR). ACRs (µg/mg) for young mice (black circles), and aged mice (white circles), at baseline prior to FSGS and at days 7, 14, 21 and 28 post-FSGS. ACR increased acutely for both young and aged animals to peak at D7, followed by gradual recovery for 21 days. Aged mice started with higher ACR at baseline and finished with significantly higher ACR at D28.

  
  
  

  

  Podocyte density was lower at baseline in aged mice, and in aged mice with FSGS. (A-C) Quantification of podocyte density. Graphs A and B show the average podocyte density in podocytes per glomerular volume (µm³) for individual animals in OC and JM glomeruli respectively. Graph C shows podocyte density for individual animals when OC and JM glomeruli are combined, which serves as a representation of the entire section. Podocyte density was lower in aged baseline mice than young baseline mice in glomeruli of the OC (A), JM (B), and when combined (C). Aged FSGS mice also had lower podocyte density in OC (A), JM (B), and combined (C) glomeruli than young FSGS mice, despite young mice experiencing a larger magnitude of podocyte depletion with FSGS. (D-G) PAS/p57 double staining. Representative images of glomeruli at 20x magnification, with higher magnifications shown in D’-G’ of the glomerulus marked by solid black square. Podocytes were identified by p57⁺ staining (brown color, nuclear) against the pink PAS counterstain.

  
  
  

  

  Parietal epithelial cell (PEC) activation was highest in aged FSGS mice. PECs were identified by PAX8 staining (green color, nuclear), and the subset of PECs undergoing activation were identified by CD44 staining (red color, cytoplasmic). (A-C) Quantitation of the percentage of PECs that are activated. (A) In outer cortical (OC) glomeruli, the percentage of PECs that were activated (PAX8⁺CD44⁺) along Bowman’s capsule (BC) was higher in aged mice at baseline and in FSGS. (B) In juxta-medullary (JM) glomeruli, the percentage of activated PECs were similar at baseline, but higher in aged FSGS mice compared to young FSGS mice. (C) When OC and JM glomeruli were combined, aged FSGS mice had the highest percentage of PECs that were activated. (D-G) Representative images of Pax8 and CD44 staining on BC. Images taken at 400x by confocal microscopy for PAX8 (green, nuclear), CD44 (red, cytoplasmic) and DAPI (blue, nuclear) staining. (D’-G’) Higher magnification of the white square shown above. Solid arrow shows examples of PAX8 staining; dashed arrow shows examples of CD44 below (D’-G’). As shown in the above graphs, the percentage of PECs along Bowman’s capsule that are activated was higher in aged baseline mice, and increased further at D28 of FSGS in aged mice.

  
  
  

  

  Activated PECs migrated from Bowman’s capsule to the glomerular tuft in FSGS. (A-C) Quantitation showing the percentage of glomeruli with activated PECs (PAX8⁺CD44⁺) on the glomerular tuft. Aged FSGS mice had the highest percentage of activated PECs on the tufts of outer cortical (OC) (A), juxta-medullary (B) and combined OC and JM (C) glomeruli. (D-G) Representative images of Pax8 and CD44 staining on tuft. Images of glomeruli (400x) taken by confocal microscopy, showing staining for PAX8 (PEC marker, green, solid arrows), CD44 (activation marker, red, dashed arrows) and DAPI (nuclei, blue). Glomeruli are marked by the dashed line. (D’-G’) Higher power images of the area demarcated by the solid square shown above. PAX8 staining was detected along Bowman’s capsule in young baseline (D, D’) and aged baseline (E, E’) mice, but not in the glomerular tuft. (F, F’) In young FSGS mice, activated PECs were not readily detected on glomerular tufts. (G, G’) Activated PECs were detected on the glomerular tuft of aged FSGS mice. These results show that activated PECs were detected on the tuft of a subset of aged FSGS glomeruli.

  
  
  

  

  Percentage of glomeruli with phosphorylated-ERK stained PECs was highest in aged baseline mice with FSGS. (A-C) Quantitation showing the percentage of glomeruli with pERK staining of PECs along Bowman’s capsule. Aged mice and aged mice with FSGS had a higher percentages of glomeruli with pERK staining along Bowman’s capsule when compared to their respective young baseline and young FSGS mice in outer cortical glomeruli (OC) (A), juxta-medullary glomeruli (B) and combined OC and JM glomeruli(C). Overall, aged FSGS mice had the highest percentage of glomerular with pERK staining (C). (D-G) Representative images of pERK staining along Bowman’s capsule. Representative images of glomeruli at 100x magnification, with 400x magnifications shown in D’-G’ of the glomeruli marked by solid black square. Dashed arrows indicate pERK negative and solid arrows indicated pERK positive glomeruli (100x) and PECs (400x).

  
  
  

  

  EMT marker staining along Bowman’s capsule was highest in aged FSGS mice. (A-C) Quantitation showing the percentage of glomeruli with α-SMA (EMT marker) staining along BC. There was no significant difference in young and aged mice at baseline in OC (A), JM (B), or combined (C) glomeruli. In OC (A), JM (B), and combined (C) glomeruli, α-SMA staining increased with disease in aged animals, while only in the JM (B) was α-SMA staining significantly increased in young mice, likely due to large variation within the sample groups. (D-G) Representative images of glomeruli with alpha-SMA staining along BC taken at 40x. Frequency and intensity α-SMA staining increased with disease (D vs. F, E vs. G), despite similar levels between young and aged animals at baseline (D vs. E). (D’-G’) Higher power images of the area demarcated by the solid square shown above, emphasizing the increase in α-SMA staining of cells along BC (F’, G’).

  
  
  

  

  Extracellular matrix accumulation was higher in Bowman’s capsule of aged FSGS mice. (A-L) Collagen IV (Col IV) staining. Representative images taken at 40x of Col IV staining (brown color) along Bowman’s capsule only (A-D, solid arrows), glomerular tuft only (E-H, dashed arrows), or along Bowman’s capsule and the glomerular tuft (I-L, solid and dashed arrows respectively). (M-X) Jones’(Silver) staining. Representative images taken at 40x of Jones’ basement membrane staining along BC only (M-P, yellow solid arrow), in the glomerular tuft only (Q-T, dashed yellow arrow), and both along BC and in the tuft (U-X, solid and dashed yellow arrows respectively) confirmed the staining patterns of Col IV.

  
  
  

• Research Paper pp undefined-undefined

  miR-557 inhibits hepatocellular carcinoma progression through Wnt/β-catenin signaling pathway by targeting RAB10

  Relevance score: 14.292585

  Xiaoye Cheng, Can Wu, Haocheng Xu, Ruixiang Zou, Taiyuan Li, Shanping Ye

  Keywords: miR-557, RAB10, hepatocellular carcinoma, epithelial-to-mesenchymal transition, Wnt/β-catenin pathway

  Published in Aging on Invalid Date

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  Accumulating evidence suggests that aberrant miRNAs participate in carcinogenesis and progression of hepatocellular carcinoma (HCC). Abnormal miR-557 expression is reported to interfere with the progression of several human cancers. However, the potential roles of miR-557 in HCC remain largely unknown. In the current study, we found that miR-557 was down-regulated in HCC tissues and cell lines, and was closely related to recurrence and metastasis of HCC. Notably, overexpression of miR-557 inhibited proliferation, migration, invasion, epithelial-to-mesenchymal transition (EMT) progression, blocked cells in G0/G1 phase of MHCC-97H cells in vitro, and suppressed tumor growth in vivo. However, loss of miR-557 facilitated these parameters in Huh7 cells both in vitro and in vivo. Moreover, RAB10 was identified as a direct downstream target of miR-557 through its 3’-UTR. Furthermore, RAB10 re-expression or knockdown partially abolished the effects of miR-557 on proliferation, migration, invasion, and EMT progression of HCC cells. Mechanistically, overexpression of miR-557 suppressed Wnt/β-catenin signaling by inhibiting GSK-3β phosphorylation, increasing β-catenin phosphorylation, and decreasing β-catenin transport to the nucleus, while knockdown of miR-557 activated Wnt/β-catenin signaling. Moreover, the TOP/FOP-Flash reporter assays showed that miR-557 overexpression or knockdown significantly suppressed or activated Wnt signaling activity, respectively. Additionally, low expression of miR-557 and high expression of RAB10 in HCC tissues was closely associated with tumor size, degree of differentiation, TNM stage and poor prognosis in HCC patients. Taken together, these results demonstrate that miR-557 blocks the progression of HCC via the Wnt/β-catenin pathway by targeting RAB10.