下调CDH17基因表达对顺铂耐药的人胃癌BGC823细胞凋亡与增殖的影响

Effect of Down-regulation of CDH17 Gene Expression on Apoptosis and Proliferation of Cisplatin-resistant Human Gastric Cancer BGC823 Cells

目的探讨下调肝肠钙粘连蛋白(CDH17)基因对顺铂耐药的人胃癌BGC823细胞凋亡和增殖的影响。方法构建si CDH17慢病毒转染的顺铂耐药人胃癌细胞株BGC823,运用噻唑蓝(MTT)实验检测细胞活力、Hoechst33258染色法和流式Annexin V-FITC/PI检测法观察细胞凋亡情况,Western blotting检测增殖凋亡通路相关蛋白的表达水平变化。结果 MTT实验表明,下调CDH17能够显著降低人胃癌BGC823细胞的存活率(P<0.05),提高人胃癌BGC823细胞对顺铂的敏感性;Hoechst33258染色法观察和Annexin V/PI双染色实验表明,下调CDH17可促进BGC823耐药细胞凋亡;Western blotting实验显示下调CDH17能够增加Bax、Cyt-c以及Cyclin D1的表达并降低Bcl-2和Bcl-x L的表达,表明CDH17可通过影响线粒体凋亡途径参与调控人胃癌BGC823细胞对顺铂的耐药性。结论下调CDH17可通过调控线粒体途径促进顺铂耐药BGC823细胞的凋亡,并调控周期蛋白肝肠钙粘连蛋白的表达来抑制耐药细胞的增殖,最终达到下调CDH17提高人胃癌BGC823细胞对顺铂的敏感性的效果。

Objective To investigate the effect of down-regulation of CDH17 gene on apoptosis and proliferation of cisplatin-resistant human gastric cancer BGC823 cells.Methods The cisplatin-resistant human gastric cancer cell line BGC823 was transfected with siCDH17 lentivirus.Cell viability was measured by MTT assay.Hoechst33258 staining and Annexin V-FITC/PI staining were used to observe the apoptosis of cells.Western blotting was used to detect the expression of apoptosis-related proteins.Results MTT assay showed that CDH17 significantly reduced the survival rate of human gastric cancer BGC823 cells(P<0.05)and enhanced the sensitivity of human gastric cancer BGC823 cells to cisplatin.Hoechst33258 staining and Annexin V/PI double staining showed that down-regulation of CDH17 could promote the apoptosis of cisplatin-resistant BGC823 cells.Western blotting showed that down-regulation of CDH17 could increase the expressions of Bax,Cyt-c and CyclinD1 and decrease the expressions of Bcl-2 and Bcl-xL,indicating that CDH17 might be involved in the regulation of cisplatin-resistance in human gastric cancer BGC823 cells through mitochondrial apoptosis pathway.Conclusion Down-regulation of CDH17 can promote the apoptosis of cisplatin-resistant BGC823 cells by regulating the mitochondrial pathway and regulate the expression of cyclin-dependent protein to inhibit the proliferation of drug-resistant cells.Finally,down-regulation of CDH17 can enhance the sensitivity of human gastric cancer BGC823 cells to cisplatin.