MiR-183调控PDCD4表达对SW1990胰腺癌细胞生长的调节作用

MiR-183 Regulates Proliferation of SW1990 Pancreatic Cancer Cell Line by Targeting PDCD4

中文关键词:  胰腺癌 MiR-183 细胞凋亡 细胞周期 PDCD4

英文关键词:Pancreatic cancer MiR-183 Apoptosis Cell cycle PDCD4

中文摘要:

      目的 观察miR-183在体外通过靶向调控程序性细胞死亡因子4（PDCD4）对SW1990胰腺癌细胞生长的调节作用。方法 将不同浓度的miR-183模拟物（miR-183 mimics）和拮抗剂（miR-183 inhibitors）通过细胞转染技术转入SW1990胰腺癌细胞株中，筛选出合适浓度，进一步应用实时荧光定量PCR（qPCR）和Western blot检测PDCD4基因和蛋白表达水平；MTT法检测miR-183 inhibitors对SW1990细胞生长的影响，流式细胞术和Hoechst染色检测SW1990细胞凋亡和细胞周期，Western blot检测抗凋亡基因bcl-2蛋白的表达。结果 细胞转染50 nmol/L miR-183 mimic和150 nmol/L inhibitor后，miR-183基因的表达量与对照比较差异有统计学意义（P<0.05）。qPCR和Western blot发现miR-183 mimics能下调PDCD4的表达，而miR-183 inhibitors能上调PDCD4的表达，下调bcl-2（P <0.05）。MTT法显示miR-183 inhibitors能抑制细胞增殖，流式细胞术和Hoechst染色显示miR-183 inhibitors能促进胰腺癌细胞的凋亡及阻滞细胞周期于G₀/G₁期。结论 miR-183拮抗剂体外能抑制胰腺癌细胞增殖，促进凋亡和阻滞细胞周期，且可能通过靶向调节PDCD4基因发挥其调节作用。

英文摘要:

      Objective To investigate the effect of miR-183 on the cell proliferation in SW1990 pancreatic cancer cell line by targeting programmed cell death factor 4(PDCD4). Methods The SW1990 pancreatic cancer cells were transfected with miR-183 mimics and inhibitors at different concentrations, the alteration of PDCD4 levels was observed at specific concentrations by qPCR and Western blot. The cellular proliferation of transfected cells was determined by MTT assay. The distribution of cell cycle and apoptosis was examined by flow cytometry (FCM) and Hoechst 33258 staining. The expression of B-cell lymphoma （bcl-2） was evaluated by Western blot. Results The miR-183 mimic and inhibitor (at concentrations of 50 nmol/L or 150 nmol/L) showed significantly increasing or decreasing effects on the levels ofmiR-183 respectively. The expression of PDCD4 was downregulated in the cells transfected with miR-183 mimics, while significantly upregulated in the cells treated with miR-183 inhibitors. Western blot showed that miR-183 inhibitors resulted in a marked decrease in the expression levels of bcl-2. The growth of SW1990 cells was obviously inhibited after anti-miR-183 treatment, while an increase of apoptosis cells proportion and cell cycle G₀/G₁ arrest were observed after miR-183 inhibitors transfection. Conclusion The miR-183 inhibitors could restrain cell proliferation, promote cell apoptosis and increase G₀/G₁ arrest in SW1990 pancreatic cancer cells, which may be possibly through targeting PDCD4.