Caspase 8和9在肿瘤局部微环境形成早期与调控蛋白Twist1的关联性

Study on the Correlation between Caspase8,9 and EMT Regulating Proteins Twist1 in the Early Stage of Tumor Microenvironment

目的：研究Caspase 8、9在肿瘤局部微环境早期线形程序性坏死（LPPCN）及血管生成拟态（VM）形成中对上皮-间充质转变（EMT）调控蛋白Twist1表达的影响。方法：将96只C57BL小鼠随机分为Caspase 8抑制剂组、Caspase 9抑制剂组、Caspase 8、9抑制剂联合用药组（联合组）和对照组,制备小鼠荷瘤模型。采用免疫组织化学法检测EMT调控蛋白Twist1在LPPCN和VM周围第1-4层肿瘤细胞的表达水平。结果：4组LPPCN周围第1-4层肿瘤细胞Twist1核阳性染色指数差异有统计学意义（P〈0.05或P〈0.01）,且呈递减趋势;对于同层肿瘤细胞,联合组和对照组的细胞核Twist1阳性染色指数比较差别有统计学意义（均P〈0.01）。LPPCN、VM周围第1-4层肿瘤细胞、同一层不同组的肿瘤细胞浆Twist1阳性染色指数差异均无统计学意义（均P〉0.05）;VM周围第1-4层肿瘤细胞、同一层不同组的肿瘤细胞核Twist1阳性染色指数差异均无统计学意义（均P〉0.05）。结论：Caspase 8、9介导的凋亡机制启动与EMT发生存在关联性。

Objective： To study the effect of linearly patterned programmed cell necrosis （LPPCN） and vasculogenic mimicry （VN） of caspase 8 and 9 on the expression of epithelial- mesenchymal transition （EMT） regulating protein Twist1 in the early stage of tumor microenvironment. Methods： Ninety-six C57BL mice were randomly divided into caspase 8 depressor group, caspase 9 depressor group, caspase 8 and 9 depressors group and the control group. The mouse tumor model was established. The expression of EMT regulating proteins Twist1 of tumor cells from the first to forth layer of LPPCN and VM was determined by immunohistochemical method. Results： There were significant differences in Twist1 nuclear expression fromⅠto Ⅳ layers of LPPCN in tumor cells between 4 groups （P 0.01）, which showed a decreasing trend. Compared with the control group, there was a significant difference in Twist1 nuclear expression in tumor cells of the same layer between caspase 8 and 9 depressors groups （P 0.01）. The positive rates of Twist1 cytoplasm expression in tumor cells fromⅠto Ⅳ layers of LPPCN and VM, or in the same layer of LPPCN and VM but not in the same group, were no significant differences （P 0.05）. The positive rates of Twist1 nuclear expression in tumor cells from Ⅰ to Ⅳ layers of VM and in the same layer of VM, but not in the same group were also no significant differences （P 0.05）. Conclusion： The mechenism of caspase 8 and 9-mediated-apoptosis is related with the occurrence of EMT.