gp96-肽复合物体外对脾淋巴细胞基因表达及其抗肿瘤效应研究  

Gene expression of spleen lymphocytes induced by gp96-peptide complexes and its anti-tumor effect in vitro

作　　者：朱昌来[1] 林琳[1] 张天一[1] 

机构地区：[1]南通大学神经再生重点实验室,江苏南通226001

出　　处：《中国病理生理杂志》2007年第9期1724-1727,共4页Chinese Journal of Pathophysiology

基　　金：江苏省教育厅高校自然科学指导计划资助项目(No.03KJD320183)

摘　　要：目的:研究H22肿瘤细胞来源的gp96-多肽复合物体外对小鼠脾淋巴细胞TNF-α、IFN-γmRNA表达的影响;并观察其培养上清诱导H22肿瘤细胞凋亡的形态学变化。方法:利用蛋白提取纯化技术提取H22肿瘤细胞来源的热休克蛋白gp96,Western blotting法鉴定所得目的蛋白;RT-PCR法检测细胞TNF-α和IFN-γmRNA的表达水平;激光共聚焦显微镜及透射电子显微镜观察细胞培养上清诱导H22肿瘤细胞的形态学变化。结果:经鉴定获得纯化的热休克蛋白gp96;gp96肽复合物诱导组的脾淋巴细胞TNF-α、IFN-γmRNA的表达强度(RV)分别为0.20±0.02及0.22±0.01,明显高于对照组的0.11±0.02、0.11±0.01和0.10±0.01、0.11±0.01(P<0.05);实验诱导组的培养上清能诱导H22肿瘤细胞凋亡的形态学改变。结论:H22肿瘤来源的热休克蛋白gp96-肽复合物能在体外增强小鼠脾淋巴细胞TNF-α、IFN-γmRNA的表达;同时,经诱导的脾细胞能分泌免疫活性物质诱导H22细胞凋亡。AIM ： To study the expressing variation of TNF-α and IFN-γ mRNA in mouse splenocytes induced by H22 tumor cells derived heat shock protein gp96 - peptide complexes in vitro, and to observe the morphologic change of H22 tumor cells which treated with the culture supernatant. METHODS ： H22 tumor cells derived HSP gp96 was obtained by the techniques for protein extraction and purification and was identified by Western blotting method. The expression values of TNF -α and IFN-γ mRNA in spleen lymphocytes were detected by semi - quantitative RT - PCR. The morphologic changes of H22 tumor cells induced by the culture supernatant were observed by laser scanning confocal microscopy （LSCM） and transmission electron microscope （TEM）. RESULTS： Purified heat shock protein gp96 was identified by Western blotting. The expression value of TNF -α and IFN -γ mRNA in activated spleen lymphocytes induced by gp96 - peptide complexes was higher than that in control groups （ P 〈 0. 05 ）. The morphologic change of apoptosis of H22 tumor cells, -which treated by the culture supernatant of experimental group was observed with LSCM and TEM. CONCLUSION： Heat shock protein gp96 - peptide complexes increase the expression value of TNF - α and IFN-γ mRNA in spleen lymphocytes of mouse in vitro. Besides, apoptosis of H22 cells is induced by immunologic active material secreted by activated splenocytes.

关 键 词：GP96 热休克蛋白质类 脾淋巴细胞 细胞凋亡 基因表达 

分 类 号：R735[医药卫生—肿瘤]