LncRNA GALNT5 uaRNA在TRAIL诱导的肺癌A549和HCC827细胞耐药中的作用

LncRNA GALNT5 uaRNA mediates the resistance to TRAIL in lung cancer A549 and HCC827 cells

目的:探究LncRNA GALNT5 uaRNA在肿瘤坏死因子相关凋亡诱导配体(TRAIL)诱导的肺癌A549和HCC827细胞耐药中的作用。方法:采用实时荧光定量PCR法分别检测正常培养及TRAIL耐药的A549和HCC827细胞中GALNT5 uaRNA的表达。A549或HCC827细胞分为4组,空载体组、GALNT5 uaRNA组、siNC组和siGALNT5 uaRNA组,转染并以TRAIL(终浓度为100g/L)处理24 h,采用双染法检测细胞凋亡,Western blot检测凋亡通路蛋白及乙酰化组蛋白H3的表达。采用RNA pull down实验验证GALNT5 uaRNA与组蛋白脱乙酰酶1(HDAC1)之间的相互作用。结果:与正常培养的细胞相比,对TRAIL耐药的A549和HCC827细胞中GALNT5 uaRNA的表达均升高(P<0.001)。与空载体组相比,过表达GALNT5 uaRNA并经TRAIL处理的A549和HCC827细胞凋亡率降低,凋亡通路蛋白Cleaved Caspase-8、Cleaved Caspase-3蛋白及乙酰化组蛋白H3的表达水平降低(P<0.001);与siNC组相比,敲低GALNT5 uaRNA并经TRAIL处理的A549和HCC827细胞凋亡率升高,Cleaved Caspase-8、Cleaved Caspase-3蛋白及乙酰化组蛋白H3的表达水平升高(P<0.001)。GALNT5 uaRNA可以与HDAC1相互作用并抑制乙酰化组蛋白H3蛋白的表达(P<0.05)。结论:GALNT5 uaRNA可能通过与HDAC1相互作用抑制组蛋白的乙酰化,从而减少Caspase-8的表达,促进肺癌细胞对TRAIL的耐药性。

Aim:To investigate the role of LncRNA GALNT5 uaRNA in tumor necrosis factor associated apoptosis inducing ligand(TRAIL)-induced drug resistance in human lung cancer A549 and HCC827 cells.Methods:The expression of GALNT5 uaRNA in normally-cultured and TRAIL-resistant A549 and HCC827 cells was detected by qRT-PCR.A549 and HCC827 cells were allocated into 4 groups,vector group,GALNT5 uaRNA group,siNC group and siGALNT5 uaRNA group,respectively,and treated with TRAIL(100 g/L) for 24 hours.Cell apoptosis was detected by double staining,and the expressions of apoptosis pathway proteins,and acetylated histone H3 were detected by Western blot.RNA pull down assay was used to verify the interaction between GALNT5 uaRNA and histone deacetylase 1(HDAC1).Results:Compared with normally-cultured cells,the expression of GALNT5 uaRNA was increased in TRAIL-resistant A549 and HCC827 cells(P<0.001).Compared with the vector group,the apoptosis rate of A549 and HCC827 cells in GALNT5 uaRNA group was decreased after TRAIL treatment,and the protein expressions of Cleaved Caspase-8,Cleaved Caspase-3,and acetylated histone H3 were also decreased(P<0.001).Compared with siNC group,the apoptosis rate of A549 and HCC827 cells in siGALNT5 uaRNA group was increased after TRAIL treatment,and the protein expressions of Cleaved Caspase-8,Cleaved Caspase-3,and acetylated histone H3 were also increased(P<0.001).GALNT5 uaRNA could interact with HDAC1,which inhibited the expressions of acetylated histone H3(P<0.05).Conclusion:GALNT5 uaRNA could inhibit histone H3 acetylation by interacting with HDAC1,thereby reducing the expression of Caspase-8 and promoting TRAIL resistance in lung cancer cells.