摘要
目的:探讨mi R-497靶向Bcl-2对肝癌细胞增殖、凋亡的调控作用。方法:选取肝癌组织及相应远癌正常组织,分别采用逆转录PCR(reverse transcriptton PCR,RT-PCR)、Western blot检测组织中mi R-497及Bcl-2蛋白表达;选取肝癌细胞系Hep G2,分别转染mi R-497 mimics及对照寡核苷酸,采用RT-PCR、Western blot检测组织中mi R-497及Bcl-2蛋白表达,采用MTT法检测各组细胞增殖活性,采用流式细胞仪检测各组细胞凋亡,采用双荧光素酶报告基因检测细胞荧光酶活性。结果:1)与远癌正常组织相比,肝癌组织中mi R-497表达明显降低(P<0.05),Bcl-2蛋白表达明显增高(P<0.05);2)与转染对照寡核苷酸相比,转染mi R-497可使Hep G2中mi R-497表达增高(P<0.05),Bcl-2蛋白表达降低(P<0.05);3)与转染对照寡核苷酸相比,共转染mi R-497与Bcl-2-WT可使Hep G2荧光素酶活性降低(P<0.05);4)转染mi R-497后,Hep G2增殖活性较转染对照寡核苷酸明显降低(P<0.05),而转染mi R-497+Bcl-2后,Hep G2增殖活性较单纯转染mi R-497明显提高(P<0.05);5)转染mi R-497后,Hep G2总凋亡比例较转染对照寡核苷酸明显增高(P<0.05);而转染mi R-497+Bcl-2后,Hep G2总凋亡比例较单纯转染mi R-497明显降低(P<0.05)。结论:mi R-497可通过靶向Bcl-2抑制肝癌细胞增殖同时促进细胞凋亡。
Objective： To evaluate the effect of miR-497 on regulating cell proliferation and apoptosis by targeting Bcl-2 in liver cancer cells. Methods： We tested liver cancer tissue and para-carcinoma tissue and used RT-PCR or Western blot to detect the expression of miR-497and Bcl-2 protein. We also tested the liver cancer cell HepG2 transfected with miR-497 mimics and mimic control. The expressions of miR-497and Bcl-2 protein were detected by RT-PCR or Western blot. Cell proliferation activity was detected by the MIT method, cell apoptosis was detected by flow cytometry, and cell luciferase activity was detected by the dual-luciferase reporter gene experiment. Results： 1） Compared with para-carcinoma tissue, the miR-497 expression of liver cancer tissue significantly decreased （P〈0.05）, whereas the Bcl-2 protein expression of liver cancer tissue significantly increased （P〈0.05）. 2） Compared with transfection mimic control, transfection miR-497 mimics could increase the miR-497 expression of liver cancer cell HepG2 （P〈0.05） and decrease the Bcl-2 protein expression of liver cancer cell HepG2 （P〈0.05）. 3） Compared with transfection mimic control, co-transfection with miR-497 mimics and Bcl-2-WT could significantly decrease the luciferase activity of liver cancer cell HepG2 （P〈0.05）. 4） Compared with transfection mimic control, the proliferative activity of liver cancer cell HepG2 significantly decreased after transfection with miR-497 mimics （P〈0.05）. Compared with transfection miR-497 mimics, the proliferative activity of liver cancer cell HepG2 significantly increased after transfection with miR-497 mimics＋Bcl-2 （P〈0.05）. 5） Compared with transfection mimic control, the total apoptosis rate of liver cancer cell HepG2 significantly increased after transfection with miR-497 mimics （P〈0.05）. Compared with transfection miR-497 mimics, the total apoptosis rate of liver cancer cell HepG2 significantly decreased after transfection with miR-497 mimics＋Bcl-2 （
出处
《中国肿瘤临床》
CAS
CSCD
北大核心
2016年第16期697-701,共5页
Chinese Journal of Clinical Oncology
