摘要
目的探讨长链非编码RNA SET结合因子2反义RNA1(lnc RNA SBF2-AS1)调控mi R-372-3p/细胞分裂蛋白激酶6(CDK6)轴对肝癌细胞侵袭及增殖的影响。方法以Bel7402和SK-hep1细胞为研究对象,上调或下调SBF2-AS1、miR-372-3p及CDK6表达水平,实时荧光定量PCR及Western blot检测细胞中miR-372-3p及CDK6表达水平。双荧光素酶报告基因实验分别验证SBF2-AS1和miR-372-3p、miR-372-3p和CDK6靶向关系。CCK-8、集落形成实验、Transwell、细胞周期实验及流式细胞术分析细胞增殖、集落形成、迁移/侵袭能力、细胞周期活动及凋亡。结果SBF2-AS1在肝癌细胞中高表达(P<0.05)。敲降SBF2-AS1后,Bel7402和SK-hep1细胞侵袭、增殖能力降低(P<0.05)。敲降miR-372-3p后,Bel7402细胞侵袭、增殖能力升高,同时敲降SBF2-AS1后,可反转上述效应(P<0.05)。miR-372-3p靶向CDK6并抑制后者的表达,过表达SFB2-AS1,可反转上述效应(P<0.05)。过表达CDK6,可以逆转过表达miR-372-3p对Bel7402细胞侵袭及增殖的抑制。结论lncRNA SBF2-AS1通过miR-372-3p正向调控CDK6的表达,改变肝癌细胞的周期活动,影响肝癌细胞的增殖、侵袭能力。
Objective To investigate the effects of lncRNA SBF2-AS1 on the proliferation and invasion of hepatoma cells by regulating the miR-372-3p/CDK6 pathway.Methods Bel7402 and SK-hep1 cells were selected as research objects.The expression levels of SBF2-AS1,miR-372-3p,and CDK6 were up-or down-regulated according to different experimental stages,while the expression levels of miR-372-3p and CDK6 in cells were detected by real-time fluorescence quantitative PCR and Western blot.Dual luciferase reporter assay verified the targeting relationships between SBF2-AS1 and miR-372-3p as well as miR-372-3p and CDK6,respectively.CCK-8,colony formation assay,Transwell,cell cycle assay,and flow cytometry were used to analyze cell proliferation,colony formation,migration/invasion ability,cell cycle activity,and apoptosis.Results SBF2-AS1 was highly expressed in hepatocellular carcinoma cells(P<0.05).SBF2-AS1 knockdown resulted in decreased proliferation and invasion of Bel7402 and SK-hep1 cells(P<0.05).After miR-372-3p knockdown,the proliferation capacity and invasion number of Bel7402 cells were significantly increased.However,the above results were reversed after SBF2-AS1 knockdown(P<0.05).In addition,miR-372-3p targeted CDK6 and inhibited its expression,although over-expressing SFB2-AS1 could reverse the above results(P<0.05).Over-expressing CDK6 could reverse the inhibition of over-expressing miR-372-3p on the proliferation and invasion of Bel7402 cells.Conclusion LncRNA SBF2-AS1 can positively regulate the expression of CDK6 through miR-372-3p.It can also influence the distribution of cell cycle and affect the proliferation and invasion abilities of hepatocellular carcinoma cells.
作者
宋巍
徐蓉
李玉鹏
李智德
王锦国
马超
孟塬
陈雄
SONG Wei;XU Rong;LI Yupeng;LI Zhide;WANG Jinguo;MA Chao;MENG Yuan;CHEN Xiong(Department of Hepatobiliary Surgery,People’s Hospital of Xinjiang Uigur Autonomous Region,Urumchi 830000,China;Department of Oncology,People’s Hospital of Xinjiang Uigur Autonomous Region,Urumchi 830000,China)
出处
《肿瘤防治研究》
CAS
2023年第7期666-674,共9页
Cancer Research on Prevention and Treatment
基金
新疆维吾尔自治区人民医院院内项目(20190114)
自治区科技厅天山创新团队计划项目(202116770)。
