慢病毒表达circLIFR对Hep3B肝癌细胞生物学行为的影响  

Effect of lentiviral expression of circLIFR on the biological behavior of Hep3B hepatocellular carcinoma cells

作　　者：李辰 刘路政 卢汉原 陈家诚 陈良 武金才 LI Chen;LIU Lu-zheng;LU Han-yuan;CHEN Jia-cheng;CHEN Liang;WU Jin-cai(Clinical College of Hainan Medical University,Haikou 571199,China;Department of Hepatobiliary and Pancreatic Surgery,Hainan Affiliated Hospital of Hainan Medical University(Hainan General Hospital),Haikou 570311,China)

机构地区：[1]海南医学院临床学院,海南海口571199 [2]海南医学院附属海南医院(海南省人民医院)肝胆胰外科,海南海口570311

出　　处：《海南医学院学报》2022年第24期1855-1859,1866,共6页Journal of Hainan Medical University

基　　金：海南省重点项目研发计划(ZDYF2020134,ZDYF2022SHFZ283);国家自然科学基金项目(81660489);海南省院士创新平台科研专项项目(YSPTZX202005);海南省研究生创新科研项目(Hys2020-355)。

摘　　要：目的:探讨慢病毒稳定高表达circLIFR对Hep3B肝癌细胞生物学行为的影响。方法:通过慢病毒包装circLIFR表达质粒,感染Hep3B细胞以构建稳定circLIFR高表达的肝癌细胞株。其中构建肝癌Hep3B细胞株感染circLIFR高表达序列的慢病毒为circLIFR高表达组,感染circLIFR高表达空载体序列的慢病毒为阴性对照组,无感染组为空白对照组。qPCR检测circLIFR的表达,Sanger测序鉴定其环状拼接位点。细胞增殖试剂盒检验细胞活力,Transwell鉴定侵袭能力。结果:qPCR和Sanger测序显示成功建立circLIFR稳定表达细胞株。circLIFR高表达组细胞增殖活力优于阴性对照组以及空白对照组,差异具有统计学意义(P<0.05)。阴性对照组、空白对照组的穿透Matrigel滤膜细胞数目依次为(270.8±18.9)个、(266.2±17.6)个,circLIFR高表达组穿透Matrigel滤膜的细胞数为(396.6±32.9)个,差异具有统计意义(P<0.05)。结论:circLIFR高表达明显提高Hep3B肝癌细胞增殖活力,促进细胞侵袭。Objective:To investigate the effect of lentiviral stable high expression of circLIFR on the biological behavior of Hep3B hepatocellular carcinoma(HCC)cells. Methods:Hep3B cell lines were infected with lentiviral packaging of circLIFR expression plasmids to construct stable circLIFR high expression HCC cells. The lentivirus infected with circLIFR high expression sequence was used as the circLIFR high expression group,the lentivirus infected with circLIFR high expression empty vector sequence was used as the negative control group,and the uninfected group was used as the blank control group. After that,circLIFR expression levels were detected by qPCR,and the back splice sites were identified by Sanger sequencing. The cell viability was examined by cell proliferation kit and invasive ability was determined by Transwell assay. Results:The qPCR and Sanger sequencing showed that the stable circLIFR expression of Hep3B cells was successfully established. The circLIFR high expression group had better cell proliferation viability than the negative control and blank control groups,and the differences were statistically significant(P<0.05). The number of cells crossing Matrigel gel in the negative control and blank control groups was(270.8±18.9)and(266.2±17.6),respectively,while the number of cells crossing Matrigel gel in the circLIFR high expression group was(396.6±32.9),and the differences were statistically significant(P<0.05). Conclusion:High circLIFR expression considerably promotes the proliferation and invasive ability of Hep3B cells.

关 键 词：环状RNA circLIFR 肝癌 生物学行为 

分 类 号：R735.7[医药卫生—肿瘤]