摘要
目的探讨LncRNA HULC通过miR-372/CXCR4轴在肝癌细胞增殖、凋亡与上皮-间质转化(EMT)中的作用。方法在线生物信息学TargetScan数据库预测靶基因;细胞转染建立基因过表达和沉默细胞模型;qRTPCR和Western blot检测基因和蛋白质表达;CCK-8分析用于细胞活力检测;细胞集落形成实验用于分析细胞增殖能力;Annexin V-FITC/PI分析用于细胞凋亡检测;免疫组织化学染色检测蛋白的阳性表达。结果在肝癌组织和肝癌细胞中,HULC和CXCR4表达上调,miR-372表达下调;TargetScan数据库分析和双荧光素酶分析揭示了miR-372与HULC或CXCR4之间的靶向关系;CCK-8分析显示HULC和CXCR4提高细胞活力,miR-372抑制细胞活力;细胞集落形成实验表明,HULC和CXCR4促进细胞增殖,miR-372抑制细胞增殖;细胞流式术结果表明,HULC和CXCR4抑制细胞凋亡,miR-372促进细胞凋亡;Western blot结果表明,HULC和CXCR4抑制E-cadherin表达,促进Vimentin表达,而miR-372促进E-cadherin表达,抑制Vimentin表达。结论HULC抑制miR-372的表达,从而促进CXCR4的表达,因此促进了肝癌细胞的增殖、抑制其凋亡并促进EMT进程。
Objective To investigate the role of LncRNA HULC in hepatoma cell proliferation,apoptosis,and epithelial-mesenchymal transition(EMT)via the miR-372/CXCR4 axis.Methods The online bioinformatics TargetScan database was used to predict target genes.Cell transfection was used to establish gene overexpression and silencing cell models.qRT-PCR and Western blot were used to detect gene and protein expression,respectively.CCK-8 assays were used to assess cell viability.Cell colony formation assays were used to analyze cell proliferation.Annexin V-FITC/PI was used to analyze apoptosis.Immunohistochemical staining was used to detect expression of proteins.Results In liver cancer tissues and cells,HULC and CXCR4 expression was upregulated,and miR-372 expression was downregulated.TargetScan database analysis and dual luciferase assays revealed a relationship between miR-372 and HULC or CXCR4.CCK-8 assays showed that HULC and CXCR4 increased cell viability,and miR-372 inhibited cell viability.Colony formation assays showed that HULC and CXCR4 promoted cell proliferation,and miR-372 inhibited cell proliferation.Flow cytometry showed that HULC and CXCR4 inhibited apoptosis,and miR-372 promoted apoptosis.Western blot analysis showed that HULC and CXCR4 inhibited the expression of E-cadherin and promoted the expression of Vimentin,while miR-372 promoted the expression of E-cadherin and inhibited the expression of Vimentin.Conclusions HULC inhibits the expression of miR-372,thereby promoting CXCR4 expression,proliferation,and EMT progression of hepatoma cells,while inhibiting apoptosis.
作者
刘远光
刘怀海
LIU Yuanguang;LIU Huaihai(Department of Hepatobiliary Surgery,Tangshan GongRen Hospital,Tangshan 063000,China;Department of General Surgery,the Second Hospital of Hebei Medical University,Shijiazhuang 050000)
出处
《中国比较医学杂志》
CAS
北大核心
2020年第10期44-55,共12页
Chinese Journal of Comparative Medicine
基金
河北省医学科学研究重点课题计划(20181504)。