摘要:背景伊立替康(camptothecin-11,CPT-11)为晚期结肠癌的一线化疗用药,但CPT-11耐药性限制了CPT-11的化疗效果.研究结肠癌CPT-11耐药的机制,并恢复结肠癌细胞对CPT-11的敏感性,对延长结肠癌患者的生命时间具有十分重要的临床价值.目的探讨长链基因间非编码RNA-p21(long intergenic noncoding RNA-p21,lincRNA-p21)对结肠癌细胞CPT-11耐药的作用和机制.方法采用结肠癌HCT-8细胞和SW480细胞利用CPT-11持续接触浓度递增法分别构建伊立替康耐药HCT-8/CPT-11细胞和SW480/CPT-11细胞,并利用实时定量聚合酶链式反应(real-time quantitative polymerase chain reaction,RT-qPCR)检测细胞内lincRNA-p21表达.HCT-8/CPT-11细胞和SW480/CPT-11细胞分别转染pcDNA-lincRNA-p21或si-lincRNA-p21后,细胞计数试剂盒-8(cell counting kit-8,CCK-8)实验检测CPT-11对细胞活性的影响;蛋白质免疫印迹(Western blot)初步分析了lincRNA-p21对磷脂酰肌醇3-激酶/蛋白激酶B(phosphoinositide 3-kinase/protein kinase B,PI3K/AKT)通路是否有调节作用.采用PI3K/AKT通路抑制剂LY294002预处理已转染si-lincRNA-p21的HCT-8/CPT-11细胞和SW480/CPT-11细胞或采用PI3K/AKT通路激动剂Recilisib预处理已转染pcDNAlincRNA-p21的HCT-8/CPT-11细胞和SW480/CPT-11细胞,CCK-8实验检测CPT-11对细胞活性的影响.结果与亲本细胞相比,CPT-11耐药细胞中lincRNA-p21表达明显降低.过表达lincRNA-p21抑制HCT-8/CPT-11细胞和SW480/CPT-11细胞对CPT-11耐药性,而敲低lincRNA-p21增强HCT-8/CPT-11细胞和SW480/CPT-11细胞对CPT-11耐药性.Western blot结果显示,lincRNA-p21过表达抑制PI3K/AKT通路活性;而敲低lincRNA-p21增强PI3K/AKT通路活性.LY294002能抑制lincRNA-p21敲低对CPT-11耐药的促进作用;Recilisib能抑制lincRNA-p21过表达对CPT-11耐药的抑制作用.结论上调lincRNA-p21能抑制结肠癌细胞CPT-11耐药性,而下调lincRNA-p21能增强结肠癌细胞CPT-11耐药性,这种调节作用可能与lincRNA-p21调控PI3K/AKT信号通路活性
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