人膀胱癌细胞株EJ中热休克蛋白70的热诱导表达及其意义

Hyperthermia-induced heat shock protein 70 expression and its significance in human bladder carcinoma cell line EJ

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。

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 in 5% CO2 incubator （37 ℃） for 2 h. The flow cytometry （FCM） and Western blot were used to determine HSP70. RT-PCR was used to determine HSP70 mRNA. 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.