LncRNA GAS5吸附miR-221-3p抑制肝细胞肝癌增殖及其启动子甲基化状态

LncRNA GAS5 inhibits the proliferation of hepatocellular carcinoma by adsorption of miR-221-3p and methylation status of its promoter region

目的:研究长链非编码RNA-生长抑制特异性基因5(growth arrest specific transcript 5,GAS5)对肝癌细胞增殖、迁移及侵袭的影响,同时研究其启动子区域的甲基化状态。方法:实时荧光定量聚合酶链式反应(real-time fluorescence quantitative polymerase chain reaction,qRT-PCR)检测GAS5在肝癌细胞株和肝癌组织中的表达,在肝癌细胞中过表达GAS5,CCK-8法检测细胞增殖,直线划痕法检测细胞迁移,Transwell小室法检测细胞侵袭。通过双荧光素酶法检测GAS5与miR-221-3p的靶向结合。通过不同剂量甲基转移酶抑制剂5-氮杂-2'-脱氧胞苷(5-Aza-2'-deoxycytidine,5-Aza-dC)的加入了解GAS5启动子区域的甲基化状态。结果:GAS5在肝癌细胞株和肝癌组织中表达显著下调。过表达GAS5明显抑制肝癌细胞增殖、迁移、侵袭及克隆形成。GAS5存在与miR-221-3p的互补结合位点。随着5-Aza-dC浓度的增加,GAS5相对表达量呈现显著上调。结论:GAS5在肝癌组织和细胞系中的表达显著下调,可能与其CpG岛甲基化有关。GAS5可作为分子海绵吸附miR-221-3p抑制肝癌细胞增殖。

Objective:To study the effect of long noncoding RNA-growth arrest specific transcript 5(GAS5)on the proliferation,migration and invasion of liver cancer cells,and to study the methylation status of its promoter region.Methods:Real-time fluorescence quantitative polymerase chain reaction(qRT-PCR)was used to detect the expression of GAS5 in liver cancer cell lines and tissues.CCK-8 method was used to detect cell proliferation,linear scratch method was used to detect cell migration,and Transwell chamber method was used to detect cell invasion.The targeted binding of GAS5 to miR-221-3p was detected by dual luciferase method.The methylation status of the GAS5 promoter region was understood by adding different doses of methyltransferase inhibitor 5-Aza-2'-deoxycytidine(5-Aza-dC).Results:GAS5 expression was significantly down-regulated in liver cancer cell lines and liver cancer tissues.Overexpression of GAS5 significantly inhibited the proliferation,migration,invasion and clonal formation of liver cancer cells.GAS5 has a complementary binding site with miR-221-3p.As the concentration of 5-Aza-dC gradually increased,the relative expression of GAS5 showed a significant increase.Conclusion:The expression of GAS5 in liver cancer tissues and cell lines is significantly down-regulated,which may be related to CpG island methylation.GAS5 can act as a molecular sponge to adsorb miR-221-3p and inhibit the proliferation of liver cancer cell.