Wang Eyou, Li Jun, Yang Guohua, Zhong Shan, Liu Tongzu
School of Basic Medical Science, Wuhan University, Wuhan, 430071, China.
Renmin Hospital of Wuhan University, Wuhan, 430060, China.
J Huazhong Univ Sci Technolog Med Sci. 2012 Apr;32(2):237-241. doi: 10.1007/s11596-012-0042-6. Epub 2012 Apr 20.
Previous researches showed that the expression level of E-Cad in most infiltrating cancer cells was reduced or negative. This study explored whether 4HPR restrained the infiltration of bladder cancer cells through regulating the expression of E-Cad. The infiltrating bladder cancer cells T24 were cultured, and then treated by a proper dosage of drug. Their viability was a determined by MTT method. Western blotting and RT-PCR were adopted to detect the changes of E-Cad gene expression at both protein and mRNA levels. Moreover, immunofluorescent staining and confocal fluorescence microscopy were employed for the observation of the expression of E-Cad. The result showed that, at both mRNA and protein levels, the expression level of E-Cad in T24 cells treated by 4HPR was significantly higher than that of control group, while the β-Cat expression was also relocated from the cell nucleus to cytoplasm. Our findings suggested that the regulatory function of 4HPR on infiltration of bladder cancer cells T24 is at least partly achieved by regulating the expression of E-Cad.
以往研究表明,大多数浸润性癌细胞中E-Cad的表达水平降低或呈阴性。本研究探讨4HPR是否通过调节E-Cad的表达来抑制膀胱癌细胞的浸润。培养浸润性膀胱癌细胞T24,然后用适当剂量的药物进行处理。采用MTT法测定其活力。采用蛋白质印迹法和RT-PCR检测E-Cad基因在蛋白质和mRNA水平的表达变化。此外,采用免疫荧光染色和共聚焦荧光显微镜观察E-Cad的表达。结果显示,在mRNA和蛋白质水平上,4HPR处理的T24细胞中E-Cad的表达水平均显著高于对照组,同时β-Cat的表达也从细胞核转移至细胞质。我们的研究结果表明,4HPR对膀胱癌细胞T24浸润的调节作用至少部分是通过调节E-Cad的表达来实现的。