CAJ | 학술논문

目的调控前列腺癌干细胞(PCSCs)中微小RNA-101(miR-101)表达,观察细胞中EZH2表达及其增殖迁移能力的变化.方法通过流式分选从LNcap细胞株中获取PCSCs;采用实时荧光定量聚合酶链反应(qRT-PCR)、Western blot、双荧光素酶、细胞凋亡、细胞周期、细胞迁移实验观察miR-101对前列腺癌干细胞EZH2表达及增殖迁移能力影响.结果在PCSCs中转染miR-101后,实验组较未处理组EZH2表达量下降46％ (P ＜0.05)；双荧光素酶实验证实PCSCs中miR-101直接调控EZH2；凋亡实验中,实验组细胞凋亡率为(3.79±0.24)％,较对照组凋亡率明显增加(P＜0.05)；周期实验中,实验组细胞G1期比例为(82.95±0.78)％,较对照组比例增加(P＜0.05),细胞被阻滞在G1/S期；迁移试验中,细胞迁移率为0.38±0.01,明显低于对照组(P＜0.05).结论过表达miR-101能下调前列腺癌干细胞中EZH2的表达并能降低细胞的增殖迁移能力.
목적 조공전렬선암간세포(PCSCs)중미소RNA-101(miR-101)표체,관찰세포중EZH2표체급기증식천이능력적변화.방법 통과류식분선종LNcap세포주중획취PCSCs;채용실시형광정량취합매련반응(qRT-PCR)、Western blot、쌍형광소매、세포조망、세포주기、세포천이실험관찰miR-101대전렬선암간세포EZH2표체급증식천이능력영향.결과 재PCSCs중전염miR-101후,실험조교미처리조EZH2표체량하강46％ (P ＜0.05)；쌍형광소매실험증실PCSCs중miR-101직접조공EZH2；조망실험중,실험조세포조망솔위(3.79±0.24)％,교대조조조망솔명현증가(P＜0.05)；주기실험중,실험조세포G1기비례위(82.95±0.78)％,교대조조비례증가(P＜0.05),세포피조체재G1/S기；천이시험중,세포천이솔위0.38±0.01,명현저우대조조(P＜0.05).결론 과표체miR-101능하조전렬선암간세포중EZH2적표체병능강저세포적증식천이능력.
Objective To study the effect of microRNA (miR)-101 on EZH2 expression,proliferation and migration of prostate cancer stem cells (PCSCs).Methods PCSCs were isolated from LNcap cell line by FACS.Quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR),Western blotting,flow cytometry and cell migration assay were used to observe the effect of miR-101 on EZH2 expression,proliferation and migration of prostate cancer stem cells.Results After transfecting miR-101 into PCSCs,the expression level of EZH2 was decreased by 46％ in the experimental group as compared with untreated group (P ＜ 0.05) ; Dual luciferase experiments confirmed miR-101 directly regulated EZH2 in PCSCs ; The apoptosis rate in the experimental group was (3.79 ± 0.24) ％,which was increased significantly (P ＜ 0.05) ; The percentage of cells in G1 phase of the experimental group was (82.95 ± 0.78) ％,which was significantly increased (P ＜ 0.05).Cells were arrested in G1/S phase;Migration capacity was 0.38 ± 0.01,which was significantly lower (P ＜ 0.05).Conclusion The overexpression of miR-101 can downregulate the EZH2 expression and inhibit proliferation and migration of PCSCs.