热休克对小鼠胚胎成纤维细胞CaM mRNA和HSF活性的影响

Effect of heat shock on expression of CaM mRNA and activity of HSF

试验分别以38、40和42℃对原代培养的小鼠胚胎成纤维细胞进行热休克处理,采用半定量PCR方法和凝胶迁移阻滞(EMSA)对CaM mRNA和HSF活性进行检测,并与对照组(37℃)比较,以确定热休克对小鼠胚胎成纤维细胞CaM mRNA和HSF活性的影响。结果表明,38℃热休克处理组小鼠胚胎成纤维细胞的CaMmRNA表达量显著高于对照组(P<0.05);40和42℃处理组细胞的CaM mRNA表达量极显著高于37℃组(P<0.01),但与40℃处理组相比,42℃处理组细胞的CaM mRNA表达量略有下降。通过凝胶阻滞分析检测HSF与DNA的结合活性,发现与对照组相比,38、40和42℃热休克处理明显增加了HSF活性。

This study was carried out to detecte the expression of CaM mRNA and the activity of HSF by using the methods of RT-PCR and Gel mobility shift assay（EMSA） after the primary cultured mouse embryonic fibroblasts were exposed to 38,40 and 42 ℃.The results showed that the expressions of CaM mRNA in 38℃ groups was significantly higher than that in the control group（P0.05）.However,the expression of CaM mRNA in the group treated at 40 and 42 ℃ were extremely higher compared to that of the control group（P0.01）;The expressions of CaM mRNA in the groups treated at 42 ℃ was lower compared with that in 40 ℃ groups.Compared to the control group,the activity of HSF in the heat shock groups was significantly increased as determined by EMSA.