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目的:探讨Slit-Robo信号对黏液表皮样癌细胞凋亡的影响及其作用机制。方法:免疫组织化学方法检测Mc3细胞Slit2及Robol蛋白表达,R5单克隆抗体作用Mc3细胞,采用FITC-AnnexinV/PI法检测细胞凋亡率,流式细胞仪检测细胞周期,DNA梯状电泳及Hoechst/PI双染法检测细胞凋亡发生,流式细胞仪细胞内抗原检测法检测Caspase-3蛋白含量。结果:Mc3细胞表达Slit2及Robol蛋白,R5抗体作用下细胞凋亡率增高,达到33.6%,形态学观察凋亡细胞明显增多并出现坏死细胞,电泳结果出现梯状条带,S期细胞明显增多,Caspase-3表达水平上调,平均荧光强度达到64%。结论:黏液表皮样癌中Slit-Robo信号可能以抑制细胞凋亡的形式参与肿瘤的形成过程,而这可能是通过Caspase-3蛋白的表达调控实现的。
Objective: To investigate the effect of Slit-Robo signal on apoptosis of mucoepidermoid carcinoma cells and its mechanism. Methods: The expression of Slit2 and Robol protein in Mc3 cells was detected by immunohistochemistry. The effect of R5 monoclonal antibody on Mc3 cells was detected by FITC-Annexin V / PI. The cell cycle was detected by flow cytometry. DNA ladder and Hoechst / PI double staining was used to detect the occurrence of apoptosis. Flow cytometry was used to detect the expression of Caspase-3 protein. Results: Slit2 and Robol proteins were expressed in Mc3 cells. The apoptosis rate of A549 cells increased by 33.6% with R5 antibody. The morphological changes of apoptotic cells were observed and the necrotic cells appeared. The results of ladder electrophoresis showed that the S phase cells were significantly increased , Caspase-3 expression increased, the average fluorescence intensity reached 64%. CONCLUSION: Slit-Robo signaling in mucoepidermoid carcinoma may be involved in tumor formation in the form of inhibiting apoptosis, which may be through the regulation of Caspase-3 protein expression.