摘要
目的探索人母系表达基因3(MEG3)在肝癌发生发展中作用及机制。方法实时定量PCR检测肝癌细胞系以及正常细胞系中MEG3表达;CCK-8检测细胞增殖;流式细胞术分析细胞凋亡;蛋白印记检测血管内皮生长因子(VEGFA)及其受体(VEGFR1)表达。结果与正常肝细胞HL-7702比较,肝癌细胞(MHCC97L、SMMC7721、MHCC97H、HepG2)中MEG3表达明显下降(P<0.01);与对照组LV-scramble比较,转染过表达载体LV-MEG3导致SMMC7721和MHCC97H细胞增殖倍数降低[SMMC7721由(5.8±0.4)倍降至(3.1±0.3)倍;MHCC97由(6.4±0.5)倍降至(3.4±0.3)倍],细胞凋亡率增加[SMMC7721由(2.8±0.4)%升至(12.4±0.6)%;MHCC97H由(2.2±0.4)%升至(13.5±0.5)%],VEGFA和VEGFR1表达下降(P<0.01)。结论长链非编码RNA MEG3可抑制肝癌细胞增殖、促进肝癌细胞凋亡,其机制可能与下调细胞内血管内皮生长因子及受体表达有关。
Objective To explore the role and mechanism of maternally expressed gene 3(MEG3)in the development of hepatocellular carcinoma.Methods The expression of MEG3 in normal cells and HL-7702 cells was detected with quantitative real-time reverse transcription PCR(qRT-PCR).Cell proliferation was assessed with Cell Counting Kit-8(CCK8)assay.Cell apoptosis was tested with flow cytometry.The expression of vascular endothelial growth factor alpha(VEGFA)and its receptor(vascular endothelial growth factor receptor 1,VEFGR1)were measured with Western blot.Results Compared with that of normal hepatic cell line HL-7702,the expression of MEG3 was decreased in hepatoma carcinoma cell lines MHCC97L,SMMC7721,MHCC97H,and HepG2(P<0.01).Compared with those of the control cells transfected with LV-scramble,the cell proliferation was attenuated in LV-MEG3-transfected SMMC7721 cells(from 5.8±0.4 to 3.1±0.3 folds)and MHCC97 cells(from 6.4±0.5 to 3.4±0.3 folds),but the cell apoptosis rate was increased(from 2.8±0.4%to 12.4±0.6%for SMMC7721 cells and from 2.2±0.4%to 13.5±0.5%for MHCC97H cells);moreover,the expressions of VEGFA and VEGFR1 were significantly inhibited in LV-MEG3-transfected SMMC7721 and MHCC97 cells(both P<0.01).Conclusion Long non-coding RNA MEG33 inhibits cell proliferation and promotes cell apoptosis in hepatocellular carcinoma cells and the effects may associate with the down regulation of VEGFA and VEFGR1.
作者
张婧婧
杨晓煜
姬颖华
张敏
李伟伟
于海川
路平
王向鹏
牛新清
ZHANG Jing-jing;YANG Xiao-yu;JI Ying-hua(Henan Provincial Collaborative Innovation Center,Institute of Medical Laboratory Science and Molecular Diagnostics and Medical Laboratory Technology,Xinxiang Medical College,Xinxiang,Henan Province 453003,China)
出处
《中国公共卫生》
CAS
CSCD
北大核心
2019年第12期1644-1647,共4页
Chinese Journal of Public Health
基金
河南省医学科技公关计划项目(201203068)
河南省自然科学基金(162300410021)