LncRNA FAS-AS1调控Fas/FasL通路对胶质瘤细胞增殖、凋亡的影响

Effects of Fas/FasL pathway regulated by LncRNA FAS-AS1 on proliferation and apoptosis of glioma cells

【目的】探讨LncRNA FAS-AS1调控凋亡相关因子/凋亡相关因子配体(Fas/FasL)通路对胶质瘤细胞U251增殖、凋亡的影响。【方法】体外培养人胶质瘤细胞株U251,将细胞分为对照组、阴性转染组、FAS-AS1转染组。实时荧光定量聚合酶链反应(qRT-PCR)检测细胞中FAS-AS1、Fas、FasL mRNA的表达。细胞计数盒8(CCK-8)法检测细胞增殖情况。Hoechst 33258染色和流式细胞术检测细胞凋亡情况。蛋白免疫印迹(WB)法检测细胞中增殖、凋亡及Fas/FasL通路蛋白的表达。【结果】与对照组比较,FAS-AS1转染组FAS-AS1、Fas、FasL的mRNA表达均显著升高(P<0.05)。与对照组比较,FAS-AS1转染组细胞活力显著降低,细胞凋亡率显著升高(P<0.05),荧光显微镜下观察细胞的形态变化,凋亡细胞出现核色质浓缩和核碎裂。与对照组比较,FAS-AS1转染组细胞蛋白Fas、FasL、凋亡蛋白Caspase-8、Caspase-3、Bax表达显著升高(P<0.05),增殖蛋白CyclinD1、CDK4、凋亡蛋白Bcl-2表达显著降低(P<0.05)。【结论】LncRNA FAS-AS1可能通过激活Fas/FasL通路抑制胶质瘤细胞U251的增殖,促进细胞凋亡,可能是胶质瘤治疗的潜在靶点。

【Objective】To investigate the effects of LncRNA FAS-AS1 on proliferation and apoptosis of glioma cell U251 by regulating apoptosis related factor/apoptosis related factor ligand pathway(Fas/FasL)pathway.【Methods】Human glioma cell line U251 was cultured in vitro and divided into control group,negative transfection group and FAS-AS1 transfection group.The mRNA expressions of FAS-AS1,Fas and FasL were detected by real-time fluorescent quantitative polymerase chain reaction(qRT-PCR).Cell proliferation was detected by cell counting box 8(CCK-8)method.Cell apoptosis was detected by Hoechst 33258 staining and flow cytometry.The expression of cell proliferation,apoptosis and Fas/FasL pathway proteins was detected by Western blot(WB).【Results】Compared with the control group,FAS-AS1 transfection group had significantly higher mRNA expression of FAS-AS1,Fas and FasL(P<0.05).Compared with the control group,FAS-AS1 transfection group had significantly lower cell viability and higher apoptosis rate(P<0.05).The morphological changes of apoptotic cells under fluorescence microscope showed that the apoptotic cells showed chromatin condensation and fragmentation.Compared with the control group,FAS-AS1 transfected group had significantly higher expression of Fas,FasL,Caspase-8,caspase-3 and Bax(P<0.05),and significantly lower expression of CyclinD1,CDK4 and Bcl-2(P<0.05).【Conclusion】LncRNA FAS-AS1 may inhibit the proliferation and promote apoptosis of U251 glioma cells by activating Fas/FasL pathway,which may be a potential target for glioma therapy.