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- Title
过表达miR⁃17⁃92 基因簇对前列腺癌细胞生物学特性的影响及机制
- Authors
周鹏; 马亮; 周珺; 徐晶晶; 刘菲; 国风
- Abstract
Objective To explore the effect and mechanism of over⁃expression of miR⁃17⁃92 gene cluster on the biological characteristics of prostate cancer cells. Methods DU145 cells were transfected with miR⁃17⁃92 gene expression plasmid and clones with stable ectopic miR⁃17⁃92 overexpression were established. The cell viabilities of DU145⁃17⁃92 and DU145⁃control cells were monitored by xCELLigence system. Cell proliferation and apoptosis were analyzed by Ki⁃67 and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. Cell cycle was detected by flow cytometry. Expression levels of proteins involved in apoptosis and Akt pathway were determined by western blotting. Results xCELLigence RTCA array data showed that the growth rate of DU145⁃17⁃92 cells was significantly higher than that of DU145⁃control cells after 24 h of seeding (P<0.01). The Ki⁃67⁃positive rates of the DU145⁃control group at 24, 48 and 72 hours were (56.57±1.68)%, (85.48±0.26)% and (90.85±2.08)%, respectively. While the Ki⁃67 positive rates of the DU145⁃17⁃92 group at the desired time points were (73.64±0.68)%, (93.43±1.23)% and (97.36±0.86)%, respectively, with a statistically significant difference at 24 hours (P<0.01). The percentages of apoptotic cells of the DU145⁃control group at 24, 48 and 72 hours were (6.76±0.09)%, (14.51±0.86)% and (20.73±1.64)%, respectively, while the apoptotic percentages of the DU145⁃17⁃92 group were (1.86±0.15)%, (7.90±0.40)% and (4.92±0.48)%, respectively. The percentages of apoptotic cells of the DU145⁃control group at different time were significantly higher than those of DU145⁃17⁃92 group (P<0.01 for all). The result of western blotting showed that the protein expression levels of Bcl⁃2 interacting mediator of cell death (BIM) and phosphatase and tensin homolog deleted on chromosome ten (PTEN) in DU145⁃control cells were 0.83±0.00 and 0.91±0.00, respectively, significantly higher than 0.16±0.00 and 0.13±0.00 of DU145⁃17⁃92 cells (both P <0.01). Overexpression of miR17⁃92 induced the phosphorylation of protein kinase B (Akt) at Ser473 while no appreciable effect on the phosphorylation of Akt at Thr308. The phosphorylated level of extracellular regulated protein kinases (ERK) in DU145⁃control cells was 0.21±0.01, significantly lower than 0.72±0.01 of DU145⁃17⁃92 cells (P<0.01). Conclusions Overexpression of miR⁃17⁃92 gene plays a pivotal role in growth, proliferation, apoptosis and cell cycle of DU145 cells through down⁃regulation of apoptotic protein BIM and tumor suppressor PTEN and activation of Akt and ERK signaling pathway.
- Publication
Chinese Journal of Oncology, 2017, Vol 39, Issue 5, p325
- ISSN
0253-3766
- Publication type
Article
- DOI
10.3760/cma.j.issn.0253⁃3766.2017.05.002