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• Research Paper Volume 11, Issue 5 pp 1333-1341

  The long non-coding RNA PTTG3P promotes growth and metastasis of cervical cancer through PTTG1

  Relevance score: 24.40449

  Xiang-cui Guo, Li Li, Zhi-hui Gao, Hong-wei Zhou, Jun Li, Qian-qing Wang

  Keywords: cervical cancer, PTTG3P, Pituitary Tumor Transforming Gene 1 (PTTG1), cancer growth and metastasis

  Published in Aging on March 10, 2019

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  The outgrowth and metastasis of cervical cancer (CC) contribute to its malignancy. Pituitary Tumor Transforming Gene 1 (PTTG1) is upregulated in many types of cancer, and enhances tumor cell growth and metastasis. However, the activation and regulation of PTTG1 in CC, especially by its pseudogene PTTG3P, have not been shown. Here, we detected significantly higher levels of PTTG1 and PTTG3P in the resected CC tissue, compared to the paired adjacent normal cervical tissue. Interestingly, the PTTG3P levels positively correlated with the PTTG1 levels. High PTTG3P levels were associated with poor patients’ survival. In vitro, PTTG1 were increased by PTTG3P overexpression, but was inhibited by PTTG3P depletion in CC cells. However, PTTG3P levels were not altered by modulation of PTTG1 in CC cells, suggesting that PTTG3P is upstream of PTTG1. Moreover, PTTG3P increased CC cell growth, likely through CCNB1-mediated increase in cell proliferation, rather than through decrease in cell apoptosis. Furthermore, PTTG3P increased CC cell invasiveness, likely through upregulation of SNAIL and downregulation of E-cadherin. Our work thus suggests that PTTG3P may promote growth and metastasis of CC through PTTG1.

  

  High PTTG1 and PTTG3P levels are detected and positively correlated in CC specimens. (A-B) The mRNA levels of PTTG1 (A) and PTTG3P (B) were examined in 10 CC specimens, compared to the paired adjacent normal cervical tissue (NCT) by RT-qPCR. Data were analyzed using Wilcoxon Test. (C) Bivariate correlations between PTTG1 and PTTG3P levels in CC were calculated by Spearman's rank correlation coefficients. (D) Kaplan-Meier curve was applied to record the overall survival of the patients that were differentiated into PTTG3P-high group and PTTG3P-low group using the cut-off point of median level of PTTG3P. *p<0.05. **p<0.01. N=10.

  
  
  

  

  PTTG3P is upstream of PTTG1 in CC. (A-B) We overexpressed PTTG1 or depleted PTTG1 (by expressing short hairpin interfering RNA for PTTG1-shPTTG1) in two CC lines, Ca-Ski and HTB31. Plasmids carrying a scrambled sequence (SCR) were used as controls. (A) RT-qPCR for PTTG1. (B) RT-qPCR for PTTG3P. (C-D) We overexpressed PTTG3P or depleted PTTG3P (by expressing short hairpin interfering RNA for PTTG3P-shPTTG3P) in two CC lines, Ca-Ski and HTB31. (C) RT-qPCR for PTTG3P. (D) RT-qPCR for PTTG1. *p<0.05 (versus SCR). NS: non-significant. N=5.

  
  
  

  

  PTTG3P promotes CC growth through increase in cell proliferation. (A-B) The effects of PTTG3P on the growth of Ca-Ski cells (A) and HTB31 (B) were assessed in an CCK-8 assay. (C-D) BrdU assay was used to measure cell proliferation, shown by quantification (C), and by representative images (D). (E-F) TUNEL assay was used to measure cell apoptosis, shown by quantification (E), and by representative images (F). *p<0.05 (versus SCR). NS: non-significant. N=5. Scale bars are 20µm.

  
  
  

  

  PTTG3P increases CC invasiveness. (A-B) The role of PTTG3P in CC cell invasiveness was assessed in a Transwell cell migration assay, shown by quantification (A), and by representative images (B). *p<0.05 (versus SCR). N=5. Scale bars are 30µm.

  
  
  

  

  PTTG3P increases CCNB1, SNAIL and inhibits E-Cadherin. (A) Representative gels for Western blot. (B) Quantification for CCNB1. (C) Quantification for SNAIL. (D) Quantification for E-Cadherin. *p<0.05 (versus SCR). N=5.