摘要
目的观察人胃癌细胞株及正常人胃黏膜上皮细胞株中miR-339-3p和miR-339-5p的表达水平,通过功能获得型(gain of function)实验验证miR-339-3p和miR-339-5p与胃癌的相关性,并阐明二者在肿瘤发生中的意义。方法采用SYBR GreenⅠ嵌合荧光法实时定量PCR技术分别检测人正常胃黏膜上皮细胞GES-1,胃癌MKN-45、SGC-7901及BGC-823细胞株中miR-339-3p和miR-339-5p的表达水平;将外源性miR-339-3p mimics和miR-339-5p mimics转染胃癌MKN-45细胞株,采用RT-PCR法检测其转染率,并应用流式细胞术和CCK-8法分别检测转染72 h后MKN-45细胞的凋亡及增殖能力的变化。结果 miR-339-3p和miR-339-5p在胃癌MKN-45、SGC-7901及BGC-823细胞株中的表达均下调;与对照组相比,转染miR-339-3p mimics及miR-339-5p mimics后,MKN-45细胞株的凋亡率明显增高(P<0.05),而增殖能力明显减弱(P<0.01)。结论 miR-339-3p和miR-339-5p在3种胃癌细胞株中均呈低表达。miR-339-3p和miR-339-5p可能参与了胃癌细胞的增殖与凋亡机理。
Objective To study the expression levels of miR-339-3p and miR-339-5p in the gastric carcinoma cell lines(SGC-7901, BGC-823, and MKN-45) and gastric surface epithelium(GES-1);detect the relationship between miR-339-3p and miR-339-5p and the gastric carcinoma cell lines in vetrio experiment through the gain of function, and further signifi cance is suggested. Methods SYBR green Ⅰ real time PCR was performed to access the expression of miR-339-3p and miR-339-5p in different cell lines(SGC-7901, BGC-823, MKN-45, and GES-1). The expression levels of miR-339-3p and miR-339-5p were verified by real time PCR experiment again after transfecting miR-339-3p mimics and miR-339-5p mimics. After that, the changes of MKN-45 cells apoptosis and proliferation at 72 h after transfection were detected by fl ow cytometry and CCK-8 method. Results The expression levels of miR-339-3p and miR-339-5p in gastric carcinoma cell lines(SGC-7901, BGC-823, and MKN-45) were down regulated. Compared with the control group, the apoptosis of MKN-45 cell line was signifi cantly higher(P<0.05), the ability of proliferation of MKN-45 cell line decreased after transfecting miR-339-3p mimics and miR-339-5p mimics within 72 hours(P<0.01). Conclusion The expression levels of miR-339-3p and miR-339-5p signifi cantly decreased in the gastric carcinoma cell lines(SGC-7901, BGC-823, and MKN-45) in contrast with gastric surface epithelium. MiR-339-3p and miR-339-5p may be involved in the apoptosis and proferation of the gastric carcinoma.
出处
《中国普外基础与临床杂志》
CAS
2014年第10期1216-1221,共6页
Chinese Journal of Bases and Clinics In General Surgery
基金
中央高校基本科研业务费专项资金小额探索项目(项目编号:lzujbky-2010-209)~~
