目的 研究川楝素(toosendanin,TSN)对人卵巢癌细胞凋亡的影响,并阐明相关机制。方法 用不同浓度TSN处理CAVO-3和A2870细胞,采用CCK-8法检测细胞存活率,比色法检测Caspase-3、Caspase-9的活性,EdU法检测细胞增殖率,Western blot法检测Bcl-2、Bax、Cyto-C蛋白的表达。结果 TSN能明显抑制CAVO-3和A2870细胞的存活率,并呈剂量(r=0.869 1,P<0.05)和时间(r=0.776 5,P<0.05)依赖性。TSN处理后A2870细胞增殖率显著下降(P<0.05),且CAVO-3和A2870细胞Caspase-3和Caspase-9的活性显著升高,并能被各自抑制剂z-DEVE-FMK和z-LEHD-FMK逆转(P<0.05),同样也逆转了CAVO-3和A2870细胞存活率(P<0.05)。TSN处理后A2870细胞中Bcl-2、Bax、Cyto-C蛋白的表达显著增加,并能被Caspase-9抑制剂z-LEHD-FMK逆转(P<0.05)。结论 TSN通过上调Caspase-3和Caspase-9的活性来激活线粒体途径,从而诱导人卵巢癌细胞凋亡,这是TSN发挥抗卵巢癌作用的机制之一。
Abstract
OBJECTIVE To study the effect of toosendanin (TSN)on the apoptosis of human ovarian cancer cell and to clarify the related mechanism. METHODS CAVO-3 and A2870 cells were treated with toosendanin of different concentrations, and CCK-8 method was used to detect the cell survival rate, colorimetric method was used to measure the activities of Caspase-3 and Caspase-9, EdU method was used to determine the cell proliferation rate, and Western blot method was used to test the expressions of Bcl-2, Bax and Cyto-C protein. RESULTS TSN could significantly inhibit the survival of CAVO-3 and A2870 cells with dose-(r=0.869 1, P<0.05) and time-(r=0.776 5, P<0.05) dependent manners. The survival rate of A2870 cell with TSN dropped significantly (P<0.05). The activities of Caspase-3 and Caspase-9 were significantly increased by TSN in CAVO-3 and A2870 cells, however, the inhibitors of Caspase-3 (z-DEVE-FMK) and Caspase-9 (z-LEHD-FMK) could reverse those effect of TSN (P<0.05) and also reverse the survival rates of CAVO-3 and A2870 cells (P<0.05). In addition, TSN could increase the expression of Bcl-2, Bax and Cyto-C protein in A2870 cells notably, which could be reversed mostly by Caspase-9 inhibitor (z-LEHD-FMK)(P<0.05). CONCLUSION TSN could induce the apoptosis in human ovarian cancer cell through up-regulating the activities of Caspase-3 and Caspase-9, and then activates the mitochondrial pathway.
关键词
川楝素 /
卵巢癌 /
线粒体途径 /
凋亡
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Key words
toosendanin /
ovarian cancer /
mitochondrial pathway /
apoptosis
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中图分类号：
R965
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参考文献
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脚注
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基金
衢州市科技计划项目资助项目(2016Y018);衢职院病毒致瘤研究科技创新团队资助项目
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