紫杉醇诱导人类原代胃癌细胞凋亡及其与细胞周期关系的研究

Induction of Apoptosis of Fresh Gastric Tumor Cells with Paclitaxel and Its Association with Cell Cycle

目的探讨紫杉醇作用于人的胃癌实体瘤的机制,为紫杉醇应用于胃癌化疗提供相应的实验室依据。方法应用流式细胞术的AnnexinⅤ/PI技术检测紫杉醇诱导的37例人胃癌原代细胞凋亡;Sub-G1方法分析细胞周期及检测晚期凋亡;琼脂糖凝胶电泳检测细胞凋亡;Confocal检测凋亡发生的形态。结果AnnexinⅤ/PI法和Sub-G1法均能检测到不同浓度紫杉醇在不同时间诱导的胃癌细胞凋亡,同一浓度在不同时间诱导的胃癌细胞凋亡与对照相比差异均有显著性意义(均P<0.01);周期分布图可发现紫杉醇作用12h出现明显的G1期细胞减少,而S和G2/M期细胞增多;25.50μg/ml药物作用12和24h琼脂糖凝胶电泳均可见典型的梯形改变;共聚焦显微成像可见紫杉醇诱导的人胃癌细胞的典型早期和晚期凋亡图像。结论紫杉醇能诱导人胃癌细胞凋亡可能是其对胃癌化疗有效的机制之一。

Objective To investigate the mechanism of paclitaxel in the treatment of fresh human gastric cancer. Methods The specimen was collected from 37 patients with advanced gastric cancer after surgery. Apoptosis rate was examined by Annexin V/PI assay and Sub-G1 of flow cytometric analyses and DNA gel electrophoresis were used to detect the activity of apoptosis. Morphologic examination was done by post-sorting confocal（PSC） analysis. Results Flow cytometry analysis revealed significantly greater increases in apoptotic cells after treatment with paclitaxel. Sub-G1 analysis indicated a pronounced decrease in the G1 phase and an increase in the S phase and G2/M phase after exposure to paclitaxel for 12 h. DNA fragmentation and PSC analyses revealed conspicuous changes of apoptosis. Conclusion Paclitaxel could effectively treat gastric cancer possibly by inducing apoptosis.