【摘要】 目的应用热休克人胶质瘤细胞的方法,观察热休克蛋白70(HSP70)在人胶质瘤细胞耐药过程中的作用。方法经43℃热处理2h的人胶质瘤细胞,应用半定量逆转录聚合酶链反应(RT-PCR)及免疫组化技术检测HSP70mR-NA及蛋白的表达情况;应用hoechst33258荧光染色的方法观察化疗药物阿霉素(adriamycin,ADM)作用后,不同处理组人胶质瘤细胞的凋亡情况。结果HSP70免疫组化,HS+ADM组阳性率为(61.5±18.3)%,与HS组(61.0±14.1)%比较没有统计学意义,但显著高于对照组(8.50±4.09)%(P<0.05),RT-PCR结果与免疫组织化学一致;ADM组凋亡细胞比率为(52.7±19.1)%,显著高于对照组(6.5±3.6)%(P<0.05),HS+ADM组(25.0±10.7)%高于对照组但显著低于ADM组(P<0.05),HS组(7.2±3.6)%与对照组没有明显差异(P>0.05)。结论热休克的方法可以诱导人胶质瘤细胞HSP70的表达;HSP70可能是引起人胶质瘤细胞耐受ADM的机制之一。更多还原

【Abstract】 Objective To investigate the expression of HSP 70 in human glioma cells by heat shocking human glioma cells SHG-44. Methods After SHG-44 cells were cultured in the condition of 43 ℃ for 2 h and then 37 ℃ for 10 h, they were cultured with adriamycin (HS+ADM, 25 ng/ml) or without (HS). SHG-44 cells getting no heat shocking and with or without adriamycin treatment served as ADM group and control. HSP 70 was examined by means of immunohistochemistry and its mRNA level was measured by RT-PCR. Results The HSP 70 positive rate was (61.5±18.3)% in HS+ADM group, with no significance to (61.0±14.1)% of HS group , but was significantly higher than control . RT-PCR analysis indicated similar outcomes. The apoptotic ratio was (52.7±19.1)% in ADM group and (25.0±10.7)% in HS+ADM group, both significanty differences were observed between these 2 groups (P<0.05) and in them with control . There was no significant difference in comparison of HS group and control. Conclusion Heat shock can induce the expression of HSP 70, indicating it may be involved in the drug resistance to ADM of human glioma.更多还原