新型共刺激分子B7-H4表达抑制对人肝癌细胞增殖和凋亡的影响  

Effects of inhibition of novel costimulatory molecule B7-H4 on proliferation and apoptosis of human hepatoma cells

作　　者：何涛[1] 胡洪[1] 谢楠 钱程 李春满 唐波 付必莽 李文 HE Tao;HU Hong;XIE Nan;QIAN Cheng;LI Chunman;TANG Bo;FU Bimang;LI Wen(Yuxi People＇ s Hospital,Yuxi 653100,China)

机构地区：[1]玉溪市人民医院,云南玉溪653100 [2]昆明医科大学第二附属医院

出　　处：《山东医药》2018年第29期33-36,共4页Shandong Medical Journal

基　　金：云南省医疗卫生单位内设研究机构科研项目(2017NS281;2017NS282)

摘　　要：目的探讨新型共刺激分子B7-H4表达抑制对人肝癌细胞增殖和凋亡的影响。方法从人肝癌细胞系Hep3B,SMMC-7721,Huh-7,PLC/PRF/5,Hep G2,HCCLM3中筛选高表达B7-H4的细胞。培养高表达B7-H4的人肝癌细胞,将所培养的细胞随机分为三组:对照组、shRNA-1组、shRNA-2组。shRNA-1组、shRNA-2组分别转染对应的B7-H4慢病毒抑制基因序列,对照组加转染试剂但不予干扰序列。采用Western blotting法检测B7-H4表达量,挑选出shRNA-2组HCCLM3肝癌细胞进行下一步实验。分别于转染1、2、3、4、5 d,CCK-8法检测肝癌细胞增殖情况;转染后72 h,流式细胞术检测肝癌细胞凋亡率。结果 6个细胞株中,HCCLM3的B7-H4相对表达量最高,为高表达B7-H4细胞株。shRNA-1组、shRNA-2组B7-H4被抑制,且shRNA-2组抑制更明显。与对照组同时间比较,除转染1 d细胞增殖活性差异无统计学意义外,shRNA-2组HCCLM3细胞增殖活性降低(P均<0.05);细胞凋亡率升高(P均<0.05)。结论抑制B7-H4表达可明显抑制人肝癌细胞的增殖活性,并促使其凋亡。Objective To investigate the effects of inhibiting novel costimulatory molecule B7-H4 expression on the proliferation and apoptosis of human hepatoma cells. Methods Cells with high expression of B7-H4 were screened out from human hepatoma cell lines Hep3 B,SMMC-7721,Huh-7,PLC/PRF/5,HepG2,and HCCLM3. Human hepatoma cells with high expression of B7-H4 were cultured,and the cultured cells were randomly divided into three groups： the control group,shRNA-1 inhibition group,and shRNA-2 inhibition group. The cells in the shRNA-1 and shRNA-2 groups were transfected with the corresponding B7-H4 lentiviral suppression gene sequences,and the cells in the control group were transfected with the reagents without interference. The expression of B7-H4 was detected by Western blotting,and the HCCLM3 hepatoma cells from the shRNA-2 group were selected for further experiments. The proliferation of human hepatoma cells was detected by CCK-8 at 1,2,3,4,and 5 days after transfection. Flow cytometry was used to detect the apoptosis of hepatoma cells. Results Among the 6 cell lines,the relative expression of B7-H4 in the HCCLM3 cells was the highest. In the shRNA-1 group and the shRNA-2 group,B7-H4 was inhibited,and the inhibition of shRNA-2 group was more significant. Compared with the control group,except the proliferative activity on day 1 after transfection,the proliferation of HCCLM3 cells in the shRNA-2 group decreased,and the apoptosis rate increased（all P 〈 0. 05）. Conclusion Inhibition of B7-H4 expression can significantly inhibits the proliferation of human hepatoma cells and promote their apoptosis.

关 键 词：肝癌 新型共刺激分子B7-H4 细胞增殖 细胞凋亡 

分 类 号：R735.7[医药卫生—肿瘤]