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目的探讨从人肺腺癌传代细胞系GLC-82细胞提纯热休克蛋白70用以制备肿瘤特异性抗原。方法培养GLC-82细胞,43℃30 min热休克诱导HSP表达,低温高速离心、DEAE-52、Sephdex G-200层析法提纯HSP-70,十二烷基磺酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)和ELISA法进行定量及定性检测。结果未经过热休克处理的GLC-82细胞提取HSP-70得率约为57μg/ml,适当的热休克处理后GLC-82细胞HSP-70得率约为75μg/ml。升高温度或延长时间均会导致细胞凋亡。结论热休克处理可以增加肿瘤细胞HSP-70的表达。
Objective To investigate the purification of heat shock protein 70 from human lung adenocarcinoma cell line GLC-82 for the preparation of tumor-specific antigens. Methods GLC-82 cells were cultured and heat-shock induced the expression of HSP at 43 ℃ for 30 min. High-speed centrifugation was performed at low temperature and high temperature. The HSP-70 was purified by DEAE-52 and Sephdex G-200 chromatography. Sodium dodecyl sulfate-polyacrylamide gel Electrophoresis (SDS-PAGE) and ELISA method for quantitative and qualitative detection. Results The yield of HSP-70 in GLC-82 cells without heat shock treatment was about 57μg / ml. The yield of HSP-70 in GLC-82 cells after heat shock treatment was about 75μg / ml. Elevated temperature or prolonged time can lead to apoptosis. Conclusion Heat shock treatment can increase the expression of HSP-70 in tumor cells.