摘要
目的探讨微小RNA-128(miR-128)对肝癌细胞增殖、凋亡的影响和机制。方法生物信息学结合双荧光素酶报告基因分析miR-128下游靶基因。肝癌细胞MHCC97H分为miR-128过表达组和阴性对照组,分别感染miR-128和阴性对照序列慢病毒。miR-128过表达稳定细胞系再次分为miR-128+空质粒组和miR-128+高尔基体蛋白73(GP73)组,感染包装对照和GP73基因的腺相关病毒。活细胞计数检测试剂盒检测各组细胞增殖,脱氧核糖核昔酸末端转移酶介导的缺口末端标记法(TUNEL)检测凋亡并计算凋亡指数。6-8周Balb/c裸鼠30只随机分为过表达组和对照组,各15只,分别接种miR-128过表达组和阴性对照组肝癌细胞,随后测量肿瘤体积。结果生物信息学和双荧光素酶报告基因显示GP73是miR-128靶基因。与阴性对照组比较,miR-128过表达组增殖能力明显下降,差异有统计学意义(P<0.05)。与miR-128+空质粒组比较,miR-128+GP73组增殖能力提高,差异有统计学意义(P<0.05)。与阴性对照组比较,miR-128表达组凋亡指数增加,差异有统计学意义(P<0.05。与miR-128+空质粒组比较,miR-128+GP73组凋亡指数降低,差异有统计学意义(P<0.05)。裸鼠接种肿瘤细胞第5周时,过表达组肿瘤体积为(1209±108)mm^3,低于对照组(1985±298)mm^3,差异有统计学意义(P<0.05)。结论miR-128抑制肝癌细胞增殖,促进凋亡。miR-128通过靶向调节GP73基因影响肝癌细胞增殖和凋亡。miR-128/GP73为临床肝癌治疗靶点提供新参考。
Objective To investigate the effect of microRNA-128(miR-128)on the proliferation,apoptosis on the hepatoma cells.Methods The target genes o£miR-128 were analyzed by using bioinforma-tics and dual luciferase reporter assay system.We overexpressed miR-128 in the MHCC97H cells and also infected the GP73 gene with virus in the cell.Cell proliferation was detected by cell counting kit and apoptosis was detected by TUNEL and apoptosis index was calculated.Thirty 6-8 weeks Balb/c nude mice were randomly divided into miR-128 overexpression group(n=15)and control group(n=15).Results The results of bioinformatics and dual luciferase reporter assay showed that GP73 was the target gene of miR-128.Compared with the negative control group,cell proliferation in miR-128 overexpression group was decreased significantly;compared with miR-128+empty plasmid group,cell in miR-128+GP73 group was significantly increased(P<0.05).Compared with the negative control group,the apoptosis index in miR-128 overexpression group was significantly increased(P<0.05).Compared with miR-128+empty plasmid group,the apoptosis index of miR-128+GP73 group was significantly increased(P<0.05).Tumor volume of the overexpression group was higher than that of the control group.5 weeks later,tumor volume of the miR-128 overexpression group(1209±108)mm^3 was significantly higher than that of the control group(1985±298)mm^3,the difference was statistically significant(P<0.05).Conclusion miR-128 can promote the prolife eration and inhibit the apoptosis of hepatoma cells by regulating GP73,which provide a reference for the prognosis evaluation of hepatoma.
作者
李建华
韩玲
Li Jianhua;Han Ling(Department of General Surgery,Xinxiang Central Hospital,Xinxiang 453000,Henan Province,China)
出处
《中华肝胆外科杂志》
CAS
CSCD
北大核心
2020年第1期57-60,共4页
Chinese Journal of Hepatobiliary Surgery
