【摘 要】
:
目的 探讨紧密连接蛋白家族成员Occludin(OCLN)在膀胱癌中的表达情况及调控膀胱癌细胞自噬的机制和临床意义.方法 (1)分析膀胱癌中OCLN在mRNA和蛋白水平的表达变化.(2)在膀胱癌T24细胞系中,敲降或过表达OCLN后,利用蛋白质印迹法检测自噬相关蛋白LC3B,P62表达变化,同时利用qRT-PCR检测与自噬相关的ANKRD37的mRNA变化,并采用独立样本t检验的方法进行组间差异分析.结果 (1)OCLN的mRNA和蛋白表达量随着膀胱癌病理分期的升高逐渐上调;(2)通过蛋白印迹法检测分析发
【机 构】
:
中山大学附属第五医院 病理科,广东 珠海 519000
论文部分内容阅读
目的 探讨紧密连接蛋白家族成员Occludin(OCLN)在膀胱癌中的表达情况及调控膀胱癌细胞自噬的机制和临床意义.方法 (1)分析膀胱癌中OCLN在mRNA和蛋白水平的表达变化.(2)在膀胱癌T24细胞系中,敲降或过表达OCLN后,利用蛋白质印迹法检测自噬相关蛋白LC3B,P62表达变化,同时利用qRT-PCR检测与自噬相关的ANKRD37的mRNA变化,并采用独立样本t检验的方法进行组间差异分析.结果 (1)OCLN的mRNA和蛋白表达量随着膀胱癌病理分期的升高逐渐上调;(2)通过蛋白印迹法检测分析发现,与对照组相比,在敲降T24细胞系OCLN后,shOCLN-1组和shOCLN-2组的LC3BⅡ/Ⅰ蛋白比值明显提高(P=0.0107,P=0.0008),P62蛋白的表达水平显著降低(P=0.0382,P=0.0327);相反,过表达OCLN后,LC3BⅡ/Ⅰ的比值明显降低(P=0.0238),P62蛋白的表达水平显著升高(P=0.0049);(3)通过qRT-PCR检测发现,与对照组相比,在敲降OCLN的T24细胞中,shOCLN-1组和shOCLN-2组ANKRD37的mRNA表达量下降(P=0.0108,P=0.0183),在过表达OCLN的T24细胞系中表达量显著升高(P=0.0238).结论 OCLN在膀胱癌中高表达,抑制OCLN的表达促进膀胱癌细胞的自噬,ANKRD37是OCLN调控膀胱癌自噬的重要靶基因.
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