LncRNA HNF1A-AS1在胶质瘤中的表达及功能研究

Expression and function of long noncoding RNA HNF1A-AS1 in gliomas

目的研究长链非编码RNA(long non-coding RNA,LncRNA)HNF1A-AS1在胶质瘤中的表达及其对胶质瘤细胞增殖、迁移、侵袭及凋亡的影响。方法运用实时定量PCR技术检测HNF1A-AS1在胶质瘤组织与正常脑组织以及胶质瘤细胞与正常胶质细胞之间的表达差异。利用特异小干扰RNA(siRNA)构建低表达的胶质瘤细胞系A172和U251。通过CCK-8实验、EDU实验、Transwell、流式实验研究胶质瘤细胞增殖、迁移、侵袭、细胞凋亡情况,Western blot研究与细胞迁移、侵袭及凋亡相关蛋白的表达变化。结果LncRNA HNF1A-AS1在胶质瘤组织和胶质瘤细胞系中高表达。抑制HNF1A-AS1表达后,胶质瘤细胞增殖、侵袭、迁移能力降低,细胞凋亡率增加。Western Blot结果显示:抑制HNF1A-AS1表达后,一些与细胞迁移、侵袭、凋亡相关蛋白的表达水平明显改变。结论LncRNA HNF1A-AS1在胶质瘤中高表达,抑制HNF1A-AS1表达能降低胶质瘤细胞的增殖、迁移、侵袭能力,同时增加胶质瘤细胞的凋亡比率。因此,HNF1A-AS1可以作为脑胶质瘤的潜在治疗靶点。

Objective To investigate the expression of long noncoding RNA(LncRNA)HNF1A-AS1 in glioma cells,and its role in proliferation,migration,invasion and apoptosis.Methods Real-time quantitative PCR was used to detect the differences of HNF1A-AS1 expression between glioma tissue and normal brain tissue and the differences between glioma cell lines and normal glial cell lines.Specific small interfering RNA(si RNA)was transfected to inhibit the expression of HNF1A-AS1 in U251 and A172 glioma cell lines.CCK-8 assay,EDU assay,Transwell assay and flow assay were used to evaluate cell proliferation,migration,invasion and apoptosis after HNF1A-AS1 inhibition,and Western blot assay to confirm proteins expression changes of migration,invasion and apoptosis.Results The expression level of HNF1A-AS1 was high in glioma cells.After the inhibition of LncRNA HNF1A-AS1,the proliferation,invasion and migration of glioma cells were significantly decreased,on the contrary,the apoptosis rate of the cells was obviously increased.Western blotting indicated that the knockdown of HNF1A-AS1 resulted in remarkable changes in expression of the proteins related to cell migration,invasion and apoptosis.Conclusions LncRNA HNF1A-AS1 is highly expressed in gliomas.Inhibition of HNF1A-AS1 expression can decrease the proliferation,migration and invasion of glioma cells,and promote apoptosis in glioma cell lines.Therefore,HNF1A-AS1 can function as a potential therapeutic target of gliomas.