摘要
目的明确结肠癌转移相关基因1(MACC1)在食管癌组织中的表达状况,探讨MACC1调控PI3K/AKT信号通路对食管癌细胞系Eca109增殖凋亡的影响。方法 Western blot检测食管癌组织及癌旁组织MACC1表达;将MACC1小干扰RNA(MACC1-siRNA)和阴性对照(siRNA-NC)转染人食管癌细胞系Eca109,以空脂质体转染的细胞作为对照组,RT-PCR及Western blot检测转染效果;转染后的细胞培养48h,CCK-8检测细胞增殖;流式细胞仪检测细胞凋亡;Western blot检测Cleaved caspase3、PI3K、AKT、p-AKT蛋白表达。结果 MACC1在食管癌组织中的表达高于癌旁组织(P<0.01);siRNA-NC组细胞中MACC1的mRNA和蛋白表达与对照组差异无统计学意义(P>0.05),MACC1-siRNA组细胞中MACC1的mRNA和蛋白表达低于对照组(P<0.01);siRNA-NC组细胞增殖率、凋亡率及Cleaved caspase3、PI3K、AKT、p-AKT蛋白表达与对照组差异无统计学意义(P>0.05);MACC1-siRNA组细胞增殖率低于对照组,凋亡率及Cleaved caspase3、PI3K、p-AKT蛋白表达高于对照组(P<0.01),两组之间AKT蛋白表达差异无统计学意义(P<0.01)。食管癌细胞与PI3K/AKT信号通路的特异性抑制剂LY294002作用后细胞的增殖与凋亡结果与转染MACC1-siRNA的结果一致。结论 MACC1在食管癌中高表达,沉默MACC1的表达能显著抑制人食管癌细胞系Eca109的增殖,并通过上调Cleaved caspase3蛋白表达促进细胞凋亡,其机制与PI3K/AKT信号通路的调节有关。
Objective To investigate the effect of MACC1 by regulating the PI3K/AKT signaling pathway on the proliferation and apoptosis of esophageal cancer cells. Methods The expression of MACC1 in esophageal cancer tissue and adjacent tissue were detected by Western blot. MACC1 small interfering RNA（MACC1-siRNA）and negative control（siRNA-NC） was transfected into human esophageal cancer cell line Eca109. With empty liposome transfection cells as the control group,transfected cells cultured for 48 h and cell proliferation was detected by CCK-8.Cell apoptosis were determined by flow cytometry and the expression of Cleaved caspase3,PI3K,AKT,p-AKT protein was detected by Western blot. Results The expression of MACC1 in esophageal cancer was significantly higher than that in paracancerous tissues（P〈0.01）. The expression of MACC1 protein between siRNA-NC group and the control group had no significant difference（P〉0.05）. MACC1 mRNA and protein expression in MACC1-siRNA group was significantly lower than the control group（P〈0.01）. The proliferation rate,apoptosis rate and the expression of Cleaved caspase3,PI3K p-AKT,AKT protein in siRNA-NC group no significant difference with control group（P〉0.05）. The proliferation in MACC1-siRNA group was significantly lower than the control group while the apoptosis rate and expression of Cleaved caspase3,PI3K and p-AKT protein was significantly higher than the control group（P〈0.01）,the expression of AKT in two groups had no significant difference（P〈0.01）. The proliferation and apoptosis of esophageal cancer cells aftern PI3K/AKT signaling pathway specific inhibitor LY294002 treated cells were consistent with the results of MACC1-siRNA. Conclusion The expression of MACC1 gene was higher in esophageal cancer. The expression of silent MACC1 can significantly inhibit proliferation of the human esophageal cancer cell line Eca109,and promote cell apoptosis through upregulation Cleaved caspase3 protein expression. Its mechanism is related to the regulation of PI3K/AKT signaling pathway.
出处
《宁夏医科大学学报》
2017年第7期744-750,共7页
Journal of Ningxia Medical University
