环孢素A对人胃癌BGC823细胞增殖与凋亡的影响

Effect of cyclosporin A on proliferation inhibition and apoptosis in gastric adenocarcinoma cell line BGC823

目的研究环孢素A(Cs A)对人胃癌BGC823细胞的生长增殖以及凋亡的影响。方法 MTT法检测不同浓度Cs A处理24、48、72 h后对BGC823细胞增殖抑制率;流式细胞术(FCM)检测细胞周期、活性氧(ROS)含量及细胞线粒体膜电位(△Ψm)改变;Annexin-V-FITC/PI双染法检测细胞凋亡率。结果 Cs A在5~20μmol/L浓度范围内和24~72 h时间范围对BGC823细胞增殖有显著抑制作用,与药物剂量、作用时间呈现量效和时效的依赖性;FCM结果显示Cs A浓度依耐性使细胞周期阻滞于G0/G1期,与对照组相比差异有统计学意义(F=33.45,P<0.05,P<0.01);细胞凋亡率随Cs A作用浓度增加而增加;5~20μmol/L Cs A浓度范围细胞内ROS含量显著增加(F=46.17,P<0.01),细胞线粒体膜电位明显降低。结论 Cs A对BGC823细胞有抑制增殖和诱导凋亡作用,其机制可能与阻滞细胞生长周期、细胞内ROS含量增加以及线粒体膜电位下降相关。

Abstract Objective To investigate the effect of cyclosporin A（CsA） on inhibitory proliferation and apoptosis in human gastric carcinoma cell line BGC823. Methods The proliferation inhibition of BGC823 cells was determined by M3T assay after different concentration at 24, 48, 72 h. The cell cycle arrest, intracellular reactive oxygen spe- cies （ROS） and mitochondrial membrane potential（MMP） were measured by utilizing flow cytometry（FCM） analy- sis. Annexin V-FITC/PI double staining was employed to detect cell apoptosis rate through FCM analysis. Results CsA significantly inhibited the human BGC823 cell growth in a dose-dependent manner within the concentration ranging from 5 to 20.0 Ixmol/L and time-dependent manner from 24 h to 72 h. FCM showed that CsA blocked the G0/G1 boundary of the cell cycle in a dose-dependent manner, compared with the control group, the difference was significant（ F = 33.45, P 〈0. 05, P 〈 0.01 ）. It also demonstrated that the apoptosis rate increased in accordance with the increase of CsA concentration, and the intracellular ROS generation was obviously increased（ F =46. 17 ,P 〈 0. 01 ） and MMP decreased after CsA treatment. Conclusion CsA suppressed the proliferation of BGC823 cells by inducing cell cycle arrest, and the underlying mechanism of apoptosis may be related to the increase of intracel- lular ROS content and the decrease of MMP.