摘要
目的:研究磷脂酶 C -γ1(phospholipaseC -γ1,PLC- γ1)对高温诱导成纤维细胞热休克蛋白70(heat shock protein 70,HSP 70)表达的影响。方法:以基因敲除方法建立的缺失 plcg1 基因(plcg1 / )及野生型(plcg1+/+ )小鼠胚胎成纤维细胞为模型,Western 免疫印迹 增强化学发光法检测它们在高温诱导时的HSP 70表达,并以酶联免疫吸附( ELISA)法做半定量分析,MTT法对比分析两种细胞的热耐受力。结果:两种细胞经43℃与45℃高温诱导 30 min,都能引起HSP 70合成,但plcg1 / 细胞HSP 70表达水平显著低于 plcg1+/+ 细胞,P<0 01;较低温度(41℃)不能诱导 plcg1 / 细胞 HSP 70 表达,却能引起 plcg1+/+ 细胞 HSP - 70 表达,且 pl cg1 / 细胞的热耐力亦明显低于 plcg1+/+ 细胞。结论:PLC- γ1 参与了热诱导成纤维细胞 HSP70表达的信号转导。
OBJECTIVE:To study the effect of phospholipase C-gamma 1(PLC-γ1) on expression of heat shock protein 70(HSP 70) in heat-stressed fibroblast. METHODS: Western blotting-enhanced chemiluminescence(ECL) and ELISA were used to determine and quantify heat-induced HSP 70 expression in plcg1 gene null mouse embryonic fibroblasts (plcg1 -/-) produced by gene knock-out techniques and its wild type (plcg1 +/+). Thermotolerance of the two cell lines was also detected by the MTT assay. RESULTS: HSP 70 increased rapidly either in plcg1 -/-cells or in plcg1 +/+ cells after heat stressed at 43℃ or 45℃ for 30 min, but the levels of HSP 70 were significant higher in plcg1 +/+than in plcg1 -/-cells,P<0.01. Lower temperature (41℃) did not induce HSP 70 expression in plcg1 -/-cells but the contrary for plcg1 +/ +cell line. Furthermore, hermotolerance of plcg1 -/-cells also deceased significantly compared with that of plcg1 +/+cell line.CONCLUSION:PLC-γ1 participates in heat-induced HS70 expression signal transduction.
出处
《肿瘤防治杂志》
2005年第3期180-182,共3页
China Journal of Cancer Prevention and Treatment
