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

作　　者：陆颖影[1] 曾悦[1] 郑俊媛 黄春兰[1] 王兴鹏[1] 

机构地区：[1]南京医科大学附属上海一院临床医学院消化内科,上海200080

出　　处：《四川大学学报（医学版）》2016年第5期691-696,共6页Journal of Sichuan University(Medical Sciences)

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-183mimics)和拮抗剂(miR-183inhibitors)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-183inhibitors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50nmol/L miR-183mimic和150nmol/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-183inhibitors能上调PDCD4的表达,下调bcl-2(P<0.05)。MTT法显示miR-183inhibitors能抑制细胞增殖,流式细胞术和Hoechst染色显示miR-183inhibitors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。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 （bcP2） 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 of miR-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/GI 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.

关 键 词：胰腺癌 MiR-183 细胞凋亡 细胞周期 PDCD4 

分 类 号：R735.9[医药卫生—肿瘤]