【摘要】 目的探讨热休克蛋白70(HSP70)在膀胱癌细胞株EJ中的热诱导表达及其意义。方法 106/ml EJ置于1.5 ml Eppendorf管中,将它们在不同温度水浴分别加热诱导2 h、4 h、6 h、8 h。5%CO2孵箱中(37℃)恢复2 h。流式细胞术(FCM)和蛋白印迹法检测HSP70,反转录-聚合酶链反应(RT-PCR)检测HSP70 m RNA,SPSS16.0统计软件处理数据。结果人膀胱癌细胞株EJ中HSP70随着温度的增高表达增多,差异有统计学意义(P<0.05),但温度超过43℃时,细胞死亡率增高;随着热诱导时间的延长而增多,差异有统计学意义(P<0.05),超过8 h EJ中HSP70不再增高。结论人膀胱癌细胞株EJ中HSP70热诱导表达的最适条件是43℃、6 h,这对基于HSP70膀胱肿瘤的瘤苗制作提供一定的实验基础。更多还原

【Abstract】 Objective To investigate the hyperthermia-induced heat shock protein 70(HSP70) expression and its significance in bladder carcinoma cell line EJ. Methods The 106/ml EJ was placed in 1.5 ml Eppendorf tube for water bath of different temperatures, which were induced by hyperthermia for 2 h, 4 h, 6 h, and 8 h, and recovered in5% CO2incubator(37 ℃) for 2 h. The flow cytometry(FCM) and Western blot were used to determine HSP70. RTPCR was used to determine HSP70 m RNA. The SPSS16.0 statistical software was used to process the data. Results The HSP70 expression in human bladder carcinoma cell line EJ was increased along with the increasing temperature,with significant difference(P<0.05). However, the cell mortality was increased when the temperature exceeded 43 ℃.The HSP70 expression was increased along with the increasing induction time, with significant difference( P<0.05).The HSP70 expression no longer increased at 8 h. Conclusion The optimal condition of hyperthermia-induced HSP70 expression in human bladder carcinoma cell line EJ is 43 ℃ for 6 h, which provides a certain experimental foundation for the production of tumor vaccine based on HSP70 bladder carcinoma.更多还原