摘要
目的:探讨miR-664及其靶基因CREB1在肝癌细胞和肝癌组织中的表达及其与化疗敏感性的关系。方法:双荧光素酶实验验证miR-664对CREB1的调控作用;生物数据库分析CREB1在肝组织的表达及患者的生存期;qRT-PCR检测肝癌细胞及组织标本miR-664和CREB1的表达,并分析miR-664的表达与患者临床病理参数的关系。体外细胞实验:采用脂质体瞬时转染法将miR-644 mimic和mimic NC转染至肝癌细胞株HepG2。采用MTT法、癌细胞单克隆形成、流式细胞术、划痕实验及Transwell小室实验测定细胞的增殖、凋亡、迁移和侵袭能力;Western blot检测凋亡相关蛋白Bcl-2、caspase-3、Bax的表达水平。结果:双荧光素酶实验验证了miR-664对CREB1的靶向调控作用。生物数据库显示CREB1在肝组织中的表达越高,患者生存期越短(P<0.001)。肝癌组织miR-664表达水平明显低于癌旁组织,CREB1表达水平明显高于癌旁组织(P<0.05)。体外细胞实验:与miR-control组相比,miR-664组、DDP组、DDP+miR-664组细胞增殖、迁移、侵袭能力、Bcl-2表达量均明显降低(P<0.05);细胞凋亡能力、caspase-3蛋白、miR-664表达量均显著增多(P<0.05)。与DDP组相比,DDP+miR-664组各项指标效果均更加显著,差异有统计学意义(P<0.05)。结论:miR-664在肝癌组织及细胞中表达量减少,过表达miR-664能有效抑制肝癌细胞的增殖、迁移和侵袭,且miR-664能够增强肝癌细胞对DDP化疗的敏感性,有望成为联合DDP治疗肝癌的潜在靶点。
Objective:To investigate the expressions of miR-664 and its target gene CREB1 in liver cancer cells and liver cancer tissues and their relationship with chemotherapy sensitivity.Methods:Dual luciferase experiment to verify the regulatory effect of miR-664 on CREB1;biological database analysis of CREB1 expression in liver tissues and patient survival;qRT-PCR was used to detect expressions of miR-664 and CREB1 in liver cancer and liver cancer tissue specimens,and analyze the relationship between expression level of miR-664 and clinical pathological parameters of patients.In vitro cell experiment:Transfect miR-644 mimic and mimic NC into liver cancer cell line HepG2 by liposome transient transfection method.MTT method,cancer cell monoclonal formation,flow cytometry,scratch test and Transwell chamber test were used to determine cell proliferation,apoptosis,migration and invasion ability;Western blot was used to detect apoptosis-related proteins Bcl-2,caspase-3,Bax expression levels.Results:Dual luciferase experiment verified the targeted regulation of miR-664 on CREB1.Biological database showed that the higher the expression of CREB1in liver tissue,the shorter the patients’ survival time(P<0.001).Expression level of miR-664 in liver cancer tissue was significantly lower than that of adjacent tissues,while expression level of CREB1 was significantly higher than that of adjacent tissues(P<0.05).In vitro cell experiment:compared with miR-control group,the cell proliferation,migration,invasion ability and Bcl-2 expression in miR-664 group,DDP group and DDP+miR-664 group were significantly reduced(P<0.05);apoptotic capacity,caspase-3 protein and miR-664 expression levels all increased significantly(P<0.05).Compared with DDP group,effects of all indicators in DDP+miR-664 group were more significant,the difference was statistically significant(P<0.05).Conclusion:Expression of miR-664 is reduced in liver cancer tissues and cells.Overexpression of miR-664 can effectively inhibit proliferation,migration and invasion of liver
作者
陈凯
张竹青
马涛
张学军
于爱军
刘金龙
李剑
傅华
CHEN Kai;ZHANG Zhuqing;MA Tao;ZHANG Xuejun;YU Aijun;LIU Jinlong;LI Jian;FU Hua(Affiliated Hospital of Chengde Medical College,Chengde 067000,China)
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2022年第9期1082-1088,共7页
Chinese Journal of Immunology
基金
2018年承德市科学技术研究与发展计划项目(201804A021)。
关键词
miR-664
肝癌
肝细胞癌
顺铂
化疗敏感性
miR-664
Liver cancer
Hepatocellular carcinoma
Cisplatin
Chemotherapy sensitivity
