抑制UHRF1基因表达对肝癌细胞生物学行为的影响  

Effect of UHRF1 gene inhibition on biological behavior of hepatoma cell

作　　者：杨从容[1] 王军[1] 吴凤鹏[1] 赵学涛[1] 边晨峰[2] YANG Congrong WANG Jun WU fengpeng ZHAO Xuetao BIAN Chenfeng(The Fourth Hospital of Hebei Medical University, Shijiazhuang 050011, Hebei, China The People＇ s Hospital of Ding- zhou City, Dingzhou 073000, Hebei, China)

机构地区：[1]河北医科大学第四医院,河北石家庄050011 [2]河北省定州市人民医院,河北定州073000

出　　处：《现代中西医结合杂志》2017年第12期1265-1267,1279,共4页Modern Journal of Integrated Traditional Chinese and Western Medicine

基　　金：河北省医学科学研究重点课题计划项目(20150756)

摘　　要：目的探讨抑制泛素样含PHD和环指域1[ubiquitin-like with plant homeodomain(PHD)and ringfinger domains 1,UHRF1]基因表达对肝癌MHCC-97H细胞增殖、周期分布和凋亡的影响。方法合成UHRF1基因特异性小发夹RNA(small hairpin RNA,shRNA),构建稳定干扰UHRF1基因表达的重组慢病毒LV-shUHRF1作为LV-shUHRF1组,并将其感染MHCC-97H细胞;以空载体慢病毒LV-shNC作为阴性对照组即LV-shNC组。采用蛋白质印迹法检测UHRF1蛋白在MHCC-97H细胞中表达情况,采用CCK-8法和流式细胞术检测2组MHCC-97H细胞增殖、周期分布和凋亡情况。结果蛋白质印迹法显示,LV-shUHRF1组细胞UHRF1蛋白表达量为0.018±0.010,LV-shNC组细胞UHRF1蛋白表达量为0.818±0.031,LV-shUHRF1组细胞UHRF1蛋白表达水平明显低于LV-shNC组(P<0.05)。LV-shUHRF1组细胞的增殖能力明显低于LV-shNC组(P<0.05),G0/G1期细胞比例和细胞凋亡率明显高于LV-shNC组(P均<0.05),S期细胞比例明显低于LV-shNC组(P<0.05)。结论通过慢病毒介导的RNA干扰技术抑制UHRF1的表达后可抑制MHCC-97H细胞的增殖,阻滞细胞周期进展,促进细胞凋亡,抑制肝癌的发生和发展。Objective It is to investigate the effect of RNA interference targeting the ubiquitin-like with PHD and ringfin- get domains 1 （UHRF1） gene on cell proliferation, periodic distribution, and apoptosis of human hepatoma carcinoma MHCC- 97H cells. Methods Small hairpin RNA （sbRNA） targeting UHRF1 were synthesized to construct recombinant lentiviral vec- tor LV-shUHRF1 which stable knockdown UHRF1 as LV-sbUHRF1 group, and the recombinant vector was infected into MH- CC-97H cells （empty lentivirus LV-shNC as LV-shNC group for a negative control）. The expression level of UHRF1 protein was detected by Western blotting, then the proliferation, cycle and apoptosis of MHCC-97H cells after infection were detected by cell counting kit-8 （CCK-8） and flow eytometry, respectively. Results The result of Western blotting showed that the UHRF1 protein expression level in MHCC-97H cells was 0. 018± 0. 010 after infection with lentivirus LV-shUHRF1 , and was 0. 818 ± 0. 031 after infection with LV-shNC, the levels of LV-shUHRF1 protein expression in LV-shUHRF1 group were signif- icantly lower than that in LV-shNC group （P 〈 0.05 ）. Compared with LV-shNC group, the proliferation of MHCC-97H cells in LV-shUHRF1 group was inhibited significantly （ P 〈 0.05 ） , and the proportion of cell in G0/G1 phase, the apoptosis rate of cells in LV-shUHRFI group were significantly increased （ all P 〈 0.05 ） , and the proportion of cell in S phase was significantly decreased （ P 〈 0.05 ）. Conclusion UHRF1 gene silencing by lentivirus-mediated shRNA interference can inhibit the develop- ment and progression of hepatic carcinoma by suppressing cell growth of MHCC-97H cells and causing cell cycle arrest and cell apoptosis.

关 键 词：RNA干扰 UHRF1基因 肝癌 增殖 凋亡 MHCC-97H细胞 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学]