Notch-1与顺铂诱导的胃癌细胞上皮间质转化的关系研究

Research on the Relationship between Notch- 1 and Gastric Cancer Cells Epithelial to Mesenchymal Transformation Induced by Cisplatin

目的研究化疗药物顺铂对胃癌细胞侵袭性的影响以及Notch-1及其下游转录因子Snail与胃癌细胞上皮间质转化(epithelial to mesenchymal transition,EMT)的关系。方法以顺铂作用后的人胃癌细胞MGC-803为研究对象,用MTT比色法、蛋白免疫印迹、半定量聚合酶链反应、细胞划痕实验、流式细胞术分别检测药物作用前后相关指标。结果顺铂作用后细胞逐渐向间质细胞形态转化,随着药物作用时间延长,胃癌细胞的上皮标记蛋白E-cadherin表达下调,间质标记蛋白vimentin和目的蛋白Notch-1及其下游转录因子Snail表达上调,细胞划痕实验观察发现胃癌细胞迁移能力增强,流式细胞术检测发现药物作用后凋亡率明显增加,S期细胞减少,而G_2期细胞明显增加。结论化疗药物顺铂对胃癌细胞有着明显的杀伤作用,短期内可诱导胃癌细胞发生EMT、Notch-1以及下游转录因子Snail与胃癌细胞EMT呈正相关。

Objective To find the effect of cisplatin on the invasion ability of gastric cancer cells and the relationship between Notch-1 and its downstream transcription factor Snail and gastric cancer cells epithelial mesenchymal transformation. Methods This research was based on the human gastric cancer cells MGC-803, which had been treated by cisplatin. And the related indicators before and after the treatment were examined by MTT assay, Western blot, PCR, cell scratch experiment and flow cytometry. Results The cancer cells transformed to mesenchymal cells form gradually after being treated with cisplatin, as the drug prolonged, the expression of epithelial marker protein E-cadherin decreased while mesenchymal marker protein vimentin increased, target protein Notch -1 and its downstream transcription factor Snail increased too. We found that the invasion ability of the cells had increased by means of Cell scratch experiment. Besides, the apoptosis rate of gastric cancer cells had increased obviously, and the cells in S phase had decreased while G2 phase increased. Conclusion Cisplatin had obvious killing effect on the gastric cancer cells and can also induce gastric cancer cells to EMT in the short term. There is a positive correlation between EMT and Notch-1 as well as its downstream transcription factor Snail.