环孢素A对人胃癌MGC80-3细胞周期影响以及诱导其凋亡相关机制的研究  

Effects of cyclosporin A induced cell line arrest and related mechanism of apoptosis in human gastric adenocarcinoma MGC80-3 cells

作　　者：黄军祥[1] 胡咏梅[2] 石海[3] 张金坤[1] 王敏红[4] 杨云龙[4] 莫丽亚[4] 吴永华[4] 

机构地区：[1]苏州市立医院本部消化内科,苏州215002 [2]安徽医科大学第一附属医院科研处,合肥230022 [3]安徽医科大学第一附属医院消化内科,合肥230022 [4]苏州市立医院本部老年医学科,苏州215002

出　　处：《安徽医科大学学报》2017年第2期169-173,共5页Acta Universitatis Medicinalis Anhui

基　　金：安徽省自然科学基金青年基金(编号:11040606Q24)

摘　　要：目的探讨环孢素A(CsA)对人胃癌MGC80-3细胞周期影响以及诱导其凋亡相关机制的研究。方法采用流式细胞仪检测不同浓度CsA作用MGC80-3细胞后,细胞生长周期的变化以及细胞内活性氧(ROS)产生;吖啶橙/溴化乙啶(AO/EB)染色于荧光显微镜观察CsA处理后细胞凋亡形态学的改变;Western blot法检测Caspase-3蛋白的表达变化。结果流式细胞仪检测结果显示CsA可浓度依耐性阻滞细胞周期在G0/G1期,与对照组比较差异有统计学意义(F=48.21,P<0.05,P<0.01);能显著增加细胞内ROS含量(F=90.24,P<0.01)。AO/EB染色后细胞发生肿胀、缩小甚至碎裂等典型凋亡形态学改变。Western blot分析结果表明CsA可激活细胞内Caspase-3蛋白的表达,与对照组比较差异有统计学意义(P<0.01)。结论 CsA可能通过阻滞细胞生长周期抑制MGC80-3细胞增殖,其诱导细胞凋亡的作用机制可能与细胞内ROS大量生成以及Caspase-3蛋白高水平的表达有关。Objective To investigate the effects of cyclosporin A（CsA） induced cell arrest and related mechanism in human gastric adenocarcinoma MGC80-3 cells. Methods Cell cycle arrest and intracellular reactive oxygen spe- cies （ROS） generation were detected by utilizing flow cytometry analysis after treatment with different concentrations of CsA on MGC80-3 cells. Morphological changes of apoptotic cells were observed under fluorescent microscope through acridine orange/ethidium bromide （AO/EB） staining method. Western blot was performed to measure Caspase-3 protein expression. Results Flow cytometry indicated that CsA could arrest the cell cycle in G0/G1 phase（F =48.21 ,P 〈 0. 05 ,P 〈 0.01 ） and significantly increased intracellular ROS content in a dose-dependent manner. Compared with the control group, the difference was statistically significant （ F = 90.24, P 〈 0. 01 ）. After being treated with CsA, AO/EB staining results showed that the cells of apoptosis were characterized by typical mor- phological changes such as swelling, shrinking and fragmentation. Western blot analysis also showed that CsA obvi- ously increased the expression of Caspase-3 protein （ P 〈 0. 01 ）. Conclusion CsA may inhibit proliferation of MGC80-3 cells by blocking cell growth cycle, and the mechanism of CsA induced apoptosis may be correlated with high levels of intraeellular ROS generation and increasing expression of Caspase-3 protein.

关 键 词：环孢素A MGC80-3细胞 细胞周期 细胞凋亡 CASPASE-3蛋白 

分 类 号：R329.25[医药卫生—人体解剖和组织胚胎学] R735.2[医药卫生—基础医学]