环状RNA circRNA_0006633在肝癌组织中的表达及对肝癌细胞增殖能力的影响

Expression of CircRNA_0006633 in Liver Cancer Tissues and Its Effect on Cell Proliferation

【目的】检测环状RNA circRNA_0006633在肝癌组织中的表达情况,并研究其对肝癌细胞增殖、凋亡及周期等生物学行为的调控作用与相关机制。【方法】采用RT-qPCR方法检测80例肝癌组织和对应的癌旁组织环状RNA circRNA_0006633的表达水平,同时分析其表达与临床参数的关系;采用慢病毒包载质粒(circ_0006633-pcDNA 3.1 plasmids)技术上调肝癌细胞中circRNA_0006633的表达水平,继而用荧光定量PCR、CCK-8、凋亡、周期以及蛋白印迹(Western blot)等实验,分析肝癌细胞的增殖、凋亡及周期等生物学特性,以及上皮间质转化通路相关标志物表达水平的变化。【结果】肝癌患者的肿瘤病灶中circRNA_0006633的表达水平较其配对癌旁正常组织明显降低,且其高表达水平越低,患者的预后越差。上调circRNA_0006633表达后肝癌细胞的增殖、抗凋亡均受到抑制,细胞阻滞与G2/M期,上皮间质转化通路受到调控。【结论】相较于正常组织,在肝癌组织中circRNA_0006633的表达水平较低;且circRNA_0006633可通过抑制上皮间质转化通路激活进程,降低肝癌细胞增殖能力,进而发挥促癌作用,其为肝癌的治疗提供了新思路。

【Objective】To detect the expression of circRNA_0006633 in liver cancer tissues and explore its biological regulation on cell proliferation,apoptosis and cell cycle.【Methods】Real-time quantitative polymerase chain reaction(RT-qPCR)was used to analyzed the expression level of circRNA_0006633 in liver cancer tissues and adjacent normal tissues,while the relationship between circRNA_0006633zs expression and clinicopathological parameters was further determined.Then biological effects of circRNA_0006633 overexpression by lentivirus-mediated-plasmids(circ_0006633-pcDNA 3.1 plasmids)on cell proliferation,apoptosis,and cycle were investigated by RT-qPCR,western blot,CCK8 and flow cytometry.【Results】Compared to the paired adjacent normal t*issues the expression level of circRNA_0006633 in liver cancer tissues was lower.Furthermore,the low expression circRNA_0006633 was negatively correlated with survival outcome.Upregulation of circRNA_0006633 effectively reduced cell proliferation,increased apoptosis and arrested cells at G2/M phages while the epithelial-mesenchymal transition(EMT)-related proteins were regulated as well.【Conclusion】Expression of circRNA_0006633 is lower in liver cancer tissues.circRNA_0006633 can inhibit the process of epithelial-mesenchymal transition pathway activation and reduce the proliferation of liver cancer cells,which provides new ideas for the treatment of liver cancer.