舌鳞癌细胞热处理后HSP27的表达变化

Expression of heat shock protein 27 in tongue squamous cell carcinoma cells after hyperthermia

目的观察热疗后舌鳞癌Tscca细胞中热休克蛋白27(heat shock protein 27,HSP27)的变化及其对凋亡的作用。方法常规培养的Tscca细胞分6组,对照组不加热,其余5组43℃水浴法热处理40 min后分别常规培养2、4、8、12、24 h,应用蛋白组学方法检测HSP27变化。运用空载质粒(pc DNA3)、pc DNA3-HSP27质粒转染Tscca细胞24 h,免疫印迹分析靶细胞中的HSP27表达。43℃水浴处理未转染及转染组细胞0 min、40 min,再培养24 h后流式细胞仪检测细胞凋亡率。结果加热后12 h内细胞内HSP27持续升高。HSP27质粒转染细胞中HSP27蛋白表达增强。0 min热处理,未转染组、pc DNA3转染组及HSP27质粒转染组细胞凋亡率无差异;40min热处理,未转染组与pc DNA3转染组凋亡率无差异,HSP27质粒转染组分别与未转染组及空载质粒转染组比较,均有差异(P<0.05)。结论热处理后舌鳞癌Tscca细胞中HSP27含量升高。HSP27的变化与Tscca细胞凋亡的变化有关。

Objective To investigate the expression of heat shock protein 27（ HSP27） in human tongue squamous cell carcinoma（ Tscca） cells after hyperthermia. Methods Tscca cells without heating were cultured at 37 ℃ and divided into six groups,control group without heating,five experimental groups which were heat shocked in a temperature-controlled water bath at 43 °C for 40 min and then cultured for 2h,4h,8h,12 h,24h repectively before being harvested for proteomics analysis. Tscca cells were transfected with plasmids of pc DNA3-HSP27 and pc DNA3 for 24 hours respectively. Expression of HSP27 was examined in transfected cells with immunoblotting assay. These untransfected-cells and transfected-cells were treated at 43℃ in a heating water bath for 0min and 40 min and then cultured for 24 hours before being collected for analysis of apoptosis ratio by flow cytometry. Results HSP27 expression in Tscca cells was up-regulated continually within 12 hours after heating. Expression of HSP27 in pc DNA3-HSP27-transfected Tscca cells increased more than in pc DNA3-transfected cells. Apoptosis ratio was significantly different between HSP27-transfected cells and untransfected-cells,and pc DNA-transfected cells after heating for 40min（ P〈0. 05）. There was no difference between untransfected-cells and pc DNA-transfected cells after heating for 40min（ P〉0. 05）. Conclusion HSP27 expression in Tscca cells increased after heating and HSP27 may inhibit hyperthermia-induced apoptosis.