LncRNA HULC降低PTEN甲基化及组蛋白乙酰化促进人胶质母细胞瘤细胞增殖和侵袭

LncRNA HULC promotes cell proliferation and invasion via reducing methylation of PTEN and histone acetylation in glioblastoma

目的:探讨长链非编码RNA肝癌高表达转录本(long non-coding RNA highly up-regulated in liver cancer,LncRNA HULC)降低PTEN启动子甲基化以及组蛋白乙酰化促进胶质母细胞瘤细胞增殖和侵袭的作用。方法:稳转HULC的胶质母细胞瘤细胞株U251,分为LncRNA HULC过表达组(HULC组)及空白对照组(vec组)。实时荧光定量PCR(qRT-PCR)检测各组HULC、PTEN表达水平。亚硫酸氢盐处理后测序(Bisulfite Sequencing PCR,BSP)检测PTEN启动子甲基化水平。蛋白免疫印迹(Western Blotting,WB)检测PTEN、组蛋白H3/H4、乙酰化组蛋白H3/H4表达水平。细胞增殖实验、平板克隆形成实验、划痕-愈合实验和Transwell实验分别检测各组细胞增殖、迁移和侵袭能力。结果:与vec组相比,HULC组PTEN启动子甲基化水平及组蛋白乙酰化水平较低(P<0.05),细胞的增殖、迁移和侵袭能力显著增高(P<0.05)。结论:LncRNA HULC降低PTEN启动子甲基化和组蛋白乙酰化水平,促进胶质母细胞瘤细胞的增殖和侵袭。

Objective:To explore the effects of long non-coding RNA highly up-regulated in liver cancer(LncRNA HULC)promoting cell proliferation and invasion via reducing methylation of PTEN and histone acetylation in glioblastoma.Methods:The glioblastoma cell line U251 stably transfected with HULC was divided into LncRNA HULC overexpression group(HULC group)and blank control group(vec group).Real-time quantitative PCR(qRT-PCR)was used to detect the expression levels of HULC and PTEN in each group.The methylation level of the promoter region in PTEN was observed by Bisulfite Sequencing PCR(BSP).The protein expression level of PTEN,histone H3/H4,ac-histone H3/H4 was tested by Western Blotting(WB).Cell proliferation,migration and invasion were verified respectively by CCK8 assay,colony formation assay,wound-healing assay and Transwell assay.Results:Compared with vec group,the methylation level in the promoter region of PTEN and the histone acetylation level were reduced in HULC group(P<0.05).Cell proliferation,migration and invasion were increased in HULC group,compared with vec group(P<0.05).Conclusion:LncRNA HULC promotes cell proliferation and invasion via reducing the methylation level in the promoter region of PTEN and histone acetylation in glioblastoma.