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目的探讨姜黄素对人食管癌EC9706细胞凋亡的诱导作用,热休克蛋白70(HSP70)在肿瘤细胞凋亡过程中在核基质上的变化及其与凋亡调控相关蛋白的关系。方法用细胞计数和流式细胞仪检测姜黄素对人食管癌EC9706细胞的增殖抑制作用,以光学显微镜和透射电镜观察姜黄素诱导人食管癌EC9706细胞凋亡前后的细胞结构变化,琼脂糖凝胶电泳观察人食管癌EC9706细胞凋亡前后的DNA结构变化。双向凝胶电泳和质谱鉴定分析HSP70在核基质中的存在与变化;并以Western blotting进行确证;激光扫描共焦显微镜观察HSP70在EC9706细胞凋亡过程中的定位及其与Bax、Bcl-2等基因产物的共定位关系。结果姜黄素能显著抑制人食管癌EC9706细胞增殖并诱导人食管癌EC9706细胞凋亡,双向凝胶电泳、质谱鉴定和结果发现并证实,HSP70在姜黄素处理前后的EC9706细胞核基质蛋白中的存在及其表达下调变化。激光扫描共焦显微镜观察结果显示,HSP70在EC9706细胞凋亡过程中与Bax、Bcl-2等基因产物具有共定位关系,且其共定位区域发生了变化。结论姜黄素对人食管癌EC9706细胞具有显著的凋亡诱导作用;HSP70作为一种新发现的核基质蛋白,在姜黄素诱导人食管癌EC9706凋亡过程中的表达与分布发生了显著变化。HSP70与凋亡相关基因的关系对EC9706细胞凋亡具有重要影响。
Objective To investigate the effect of curcumin on the apoptosis of human esophageal carcinoma EC9706 cells, the change of heat shock protein 70 (HSP70) in the nuclear matrix and the relationship with heat shock protein 70 (HSP70) in the apoptosis of tumor cells. Methods The proliferation of human EC9706 cells was detected by cell counting and flow cytometry. The changes of cell structure of EC9706 cells induced by curcumin were observed by light microscope and transmission electron microscope. Electrophoresis Changes in DNA Structure of Human Esophageal Cancer EC9706 Cells before and after Apoptosis. Two-dimensional gel electrophoresis and mass spectrometry identification of HSP70 in the nuclear matrix of existence and changes, and confirmed by Western blotting; laser scanning confocal microscope HSP70 in EC9706 apoptosis process and its relationship with Bax, Bcl-2 Colocalization of gene products. Results Curcumin significantly inhibited the proliferation of human EC9706 cells and induced the apoptosis of human EC9706 cells. Two-dimensional gel electrophoresis, mass spectrometry identification and results showed that the expression of HSP70 in nuclear matrix proteins of EC9706 cells before and after curcumin treatment and Its expression is down-regulated. Confocal laser scanning microscopy showed that HSP70 had a co-localization relationship with the gene products of Bax and Bcl-2 in the process of EC9706 apoptosis, and its colocalization region had changed. CONCLUSION: Curcumin can induce apoptosis of human EC9706 cells. HSP70, a newly discovered nuclear matrix protein, has a significant change in the expression and distribution of curcumin induced apoptosis of human EC9706 cells. The relationship between HSP70 and apoptosis-related genes has an important effect on the apoptosis of EC9706 cells.