丹皮酚诱导人胃癌 SGC-7901 细胞 G2/M 期阻滞和凋亡。
Paris chinensis dioscin induces G2/M cell cycle arrest and apoptosis in human gastric cancer SGC-7901 cells.
机构信息
Department of Pathology and Pathophysiology, Taishan Medical University, Taian, Shandong Province, China.
出版信息
World J Gastroenterol. 2011 Oct 21;17(39):4389-95. doi: 10.3748/wjg.v17.i39.4389.
AIM
To investigate the anti-tumor effects of Paris chinensis dioscin (PCD) and mechanisms regarding cell cycle regulation and apoptosis in human gastric cancer SGC-7901 cells.
METHODS
Cell viability was analyzed by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide assay. Cell apoptosis was evaluated by flow cytometry and laser scanning confocal microscope (LSCM) using Annexin-V/propidium iodide (PI) staining, and the cell cycle was evaluated using PI staining with flow cytometry. Intracellular calcium ions were detected under fluorescence microscope. The expression of cell cycle and apoptosis-related proteins cyclin B1, CDK1, cytochrome C and caspase-3 was measured by immunohistochemical staining.
RESULTS
PCD had an anti-proliferation effect on human gastric cancer SGC-7901 cells in a dose- and time-dependent manner. After treatment of SGC-7901 cells with PCD, apoptosis appeared in SGC-7901 cells. Morphological changes typical of apoptosis were also observed with LSCM by Annexin V/PI staining, and the cell number of the G0/G1 phase was decreased, while the number of cells in the G2/M phase was increased. Cell cycle-related proteins, such as cyclin B1 and CDK1, were all down-regulated, but caspase-3 and cytochrome C were up-regulated. Moreover, intracellular calcium accumulation occurred in PCD-treated cells.
CONCLUSION
G2/M phase arrest and apoptosis induced by PCD are associated with the inhibition of CDK-activating kinase activity and the activation of Ca(2+)-related mitochondrion pathway in SGC-7901 cells.
目的
研究贯叶连翘薯蓣皂苷（PCD）对人胃癌 SGC-7901 细胞的抗肿瘤作用及其对细胞周期调控和凋亡的机制。
方法
采用噻唑蓝（MTT）比色法检测细胞活力。流式细胞术和激光扫描共聚焦显微镜（LSCM）用 Annexin-V/PI 染色法检测细胞凋亡，流式细胞术用 PI 染色法检测细胞周期。荧光显微镜下检测细胞内钙离子。免疫组化法检测细胞周期和凋亡相关蛋白 cyclin B1、CDK1、细胞色素 C 和 caspase-3 的表达。
结果
PCD 对人胃癌 SGC-7901 细胞具有剂量和时间依赖性的增殖抑制作用。PCD 处理 SGC-7901 细胞后，SGC-7901 细胞出现凋亡。LSCM 用 Annexin V/PI 染色观察到典型的凋亡形态学改变，G0/G1 期细胞数减少，G2/M 期细胞数增加。细胞周期相关蛋白 cyclin B1 和 CDK1 均下调，而 caspase-3 和细胞色素 C 则上调。此外，PCD 处理的细胞中发生了细胞内钙积累。
结论
PCD 诱导的 SGC-7901 细胞 G2/M 期阻滞和凋亡与抑制 CDK 激活激酶活性和激活 Ca(2+)相关的线粒体途径有关。
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