摘要
研究不同热休克条件对H22小鼠肝癌细胞的HSP70mRNA及蛋白表达的影响,以期获得最佳诱导条件.分别用MTT、RT-PCR、免疫荧光和FCM检测等方法观察不同热休克条件对鼠H22细胞的生存率、胞内HSP70mRNA转录水平及胞膜HSP70蛋白表达水平的改变.结果表明,H22细胞经轻度热休克(42~43℃),细胞生存率无影响,而中重度热休克(44~45℃)则随休克时间的延长,细胞生存率明显下降;42℃热休克初期(0.5~4小时)胞内HSP70mRNA水平低于对照组,休克后期(8~12小时)mRNA水平有所恢复并逐渐增高;不同时间热休克组H22细胞胞膜HSP70表达均较对照组显著增高(P<O.005),其中非致死性热休克以12小时表达最高,42℃为59.5%,43℃为96.8%.因此适当热休克条件可引起H22细胞HSP7OmRNA的改变及膜上HSP70蛋白表达的增加,从而增加肿瘤细胞的免疫原性.
To identify the best inducing condition, we studied the expression of membrane protein HSP70 and mRNA of H22 cell at various temperature. Using MTT,RT-PCR, immunofluorescence and FCM techniques, we observed H22 cell survival rate, the expression of HSP70 mRNA and membrane HSP70. No effects of H22 cell survival rate under 42 ~ 43℃ was observed, but cell survival rate declined with increasing stress time at 44 ~ 45 ℃; the level of HSP70 mRNA decreased initially (0.5~4.0) hours but gradually resumed and increased from 8 to 12 hours at 42℃. Membrane HSP70 expressing cells were significently higher in heat shock treatment group than in a control group ( P < 0.005). The highest expressing rate was 59.5% at 42℃ and 96.8% at 43℃ after 12 hours heat treatment. Optimal heat treatment could change HSP 70 mRNA and increase expression of membrane HSP70 of H22 cell, thus probably increase immunogenicity of the tumor cells.
出处
《中国肿瘤生物治疗杂志》
CAS
CSCD
1998年第1期5-8,共4页
Chinese Journal of Cancer Biotherapy
基金
卫生部科学研究基金(96-1-160)资助