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• Research Paper Volume 12, Issue 9 pp 8289-8300

  Pluronic P123 modified nano micelles loaded with doxorubicin enhanced tumor-suppressing effect on drug-resistant breast cancer cells

  Relevance score: 17.180326

  Xiaoyu Zhang, Weibin Chen, Jie Bai, Lijun Jin, Xiaoning Kang, Hui Zhang, Zunyi Wang

  Keywords: nano micelles, multi-drug resistance, breast cancer, P-glycoprotein, Pluronic P123 modification

  Published in Aging on May 12, 2020

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  Objective: Nano micelles (NMs) have been widely used for various biomedical applications due to its unique physiochemical properties. This study aimed to investigated the anti-tumor effect of doxorubicin (Dox)-loaded Pluronic P123 (P123) and PEG2000-DSPE mixed NMs in drug-resistant breast cancer cells.

  Results: The expression of P-gp and MDR1 gene was highly expressed in MCF-7R but not MCF-7 cells. The cellular uptake of P123-PEG2000-DSPE (Dox) was higher than that of free Dox and PEG2000-DSPE (Dox) in MCF-7R cells. Furthermore, compared with free Dox, both PEG2000-DSPE (Dox) and P123-PEG2000-DSPE (Dox) significantly diminished cell viability, and promoted cell apoptosis in MCF-7R cells. In addition, the P123-modified NMs obviously inhibited the expression of P-gp and MDR1.

  Conclusions: P123-PEG2000-DSPE (Dox) had a superior anti-tumor activity than PEG2000-DSPE (Dox) in MCF-7R cells through P-gp-mediated drug excretion and drug resistance mechanisms.

  Methods: The PEG2000-DSPE NMs (PEG2000-DSPE), P123 and PEG2000-DSPE mixed NMs (P123-PEG2000-DSPE), Dox-loaded PEG2000-DSPE NMs (PEG2000-DSPE (Dox)), and Dox-loaded Pluronic P123 and PEG2000-DSPE mixed NMs (P123-PEG2000-DSPE (Dox)) were prepared, and then the morphologies and the size distribution of PEG2000-DSPE (Dox) and P123-PEG2000-DSPE (Dox) were observed by transmission electron microscopy (TEM) and dynamic light scattering (DLS), respectively.

  

  Characterization of P123-PEG2000-DSPE (Dox). (A) The morphological characteristics of P123-PEG2000-DSPE (Dox) observed by transmission electron microscope. (B) Size distributions of P123-PEG2000-DSPE (Dox) determined by dynamic light scattering. (C) Cumulative drug release profiling of PEG2000-DSPE (Dox) and P123-PEG2000-DSPE (Dox) in phosphate-buffer saline (PBS, pH 7.4 and pH 5.0).

  
  
  

  

  Expression of P-glycoprotein (P-gp) in MCF-7 and MCF-7R cells. (A) The mRNA levels of multi-drug resistance (MDR1) gene in MCF-7 and MCF-7R cells by qRT-PCR. (B) The protein expression of P-gp in MCF-7 and MCF-7R cells using western blotting. (C) The expression of P-gp in MCF-7 and MCF-7R cells by cell immunofluorescence.

  
  
  

  

  Cellular uptake of Dox, PEG2000-DSPE (Dox) or P123-PEG2000-DSPE (Dox) by MCF-7R cells. Confocal images of MCF-7R cells treated with Dox, PEG2000-DSPE (Dox) or P123-PEG2000-DSPE (Dox) at 10 μg/mL for 0.5 and 1 h.

  
  
  

  

  P123-PEG2000-DSPE (Dox) inhibited cell growth in MCF-7 and MCF-7R cells. (A) Cell viability of MCF-7 and MCF-7R cells treated with different doses of PEG2000-DSPE (unloaded nano micelles) at 24 h by MTT assay. (B) Cell viability of MCF-7 and MCF-7R cells treated with different doses of P123-PEG2000-DSPE at 24 h by MTT assay. (C) Cell viability of MCF-7 cells treated with PBS (control), Dox, PEG2000-DSPE (Dox) or P123-PEG2000-DSPE (Dox) by MTT assay. (D) Cell viability of MCF-7R cells treated with PBS (control), Dox, PEG2000-DSPE (Dox) or P123-PEG2000-DSPE (Dox) by MTT assay.

  
  
  

  

  P123-PEG2000-DSPE (Dox) and P123-PEG2000-DSPE inhibited the expression of P-glycoprotein (P-gp) in MCF-7R cells. (A) The mRNA levels of multi-drug resistance (MDR1) gene in MCF-7R cells treated with PBS (control), PEG2000-DSPE, P123-PEG2000-DSPE, Dox, PEG2000-DSPE (Dox) and P123-PEG2000-DSPE (Dox) by qRT-PCR. (B) The protein expression of P-gp in MCF-7R cells underwent various treatments using western blotting. (C) The expression of P-gp in MCF-7R cells underwent various treatments by cell immunofluorescence.

  
  
  

  

  P123-PEG2000-DSPE (Dox) induced cell apoptosis in MCF-7R cells. Cell apoptosis rate of MCF-7R cells treated with PBS (control), Dox, PEG2000-DSPE (Dox) or P123-PEG2000-DSPE (Dox) by flow cytometry analysis. **P < 0.01 and ***P < 0.001 vs. control group; ^###P < 0.001 vs. Dox group; ^&&P < 0.01 vs. PEG2000-DSPE (Dox) group.

  
  
  

  

  P123-PEG2000-DSPE (Dox) exhibited better anti-tumor effect of on drug-resistant BC mice. (A) The body weights of mice with different treatments, including saline solution (control), Dox, PEG2000-DSPE (Dox), and P123-PEG2000-DSPE (Dox), respectively, every three days for 18 days. (B) The tumor volumes of mice with different treatments, including saline solution (control), Dox, PEG2000-DSPE (Dox), and P123-PEG2000-DSPE (Dox), respectively, every three days for 18 days. (C) The tumor weights of mice with different treatments, including saline solution (control), Dox, PEG2000-DSPE (Dox), and P123-PEG2000-DSPE (Dox), respectively, on 18 days. ***P < 0.001