灵芝孢子粉抑制人卵巢癌细胞增殖及诱导凋亡的体外研究

摘要

Objective To investigate the effect of Ganoderma lucidum spore powder ( GLSP) on cell proliferation and apoptosis of human ovarian cell line-OV 2008,and to explore its action mechanism .Methods The effects of GLSP on OV2008 cell growth and cell morphous were detected by WST-1 colorimetric technique and microscopy; cell cycle was analyzed flow cytometry;cell apoptosis was detected by Hoechst-33258 staining method;the expression of apoptosis-related proteins ( bcl-2 bax,P27,cleaved Caspase-3) were measured by Western blotting .Results The results of WST-1 assays showed that GLSP could inhibit the growth of OV2008 cells,moreover,which was time and dose dependent .The results by flow cytometry showed that GLSP could block OV2008 cell growth at G1 phase.Hoechst 33258 staining results showed that GLSP could induce cell apoptosis.Western blotting results showed that the expression levels of bcl-2, bcl-xl were decreased with the increase of concentration of GLSP,however bax,P27,Caspase-3 could inhibit the growth of ovarian cancer cells and induce cell apoptosis , and its action mechanism was that the expression levels of Casbase-3 that affected cell cycle were gradually increased . Conclusion GLSP can inhibit the growth of ovarian cancer cells , block cell growth at G1 stage, furthermore, which can develop its effects by up-regulating the expression of bax , P27, down-regulating the expression of bcl-2 and activating Caspase-3.%目的：研究灵芝孢子粉对人卵巢癌OV2008细胞增殖及凋亡的影响，并探讨其作用机制。方法采用WST－1比色法及显微镜观察法研究灵芝孢子粉对OV2008细胞生长抑制作用及细胞形态的改变；流式细胞仪检测细胞周期分布情况；Hoechst－33258染色方法评估细胞凋亡并采用western blot 方法测定凋亡相关蛋白的表达。结果WST结果表明灵芝孢子粉可以抑制OV2008细胞的生长，并呈剂量和时间依赖性；流式细胞术结果提示灵芝孢子粉可使卵巢癌细胞生长阻滞于G1期。 Hoechst－33258染色结果显示灵芝孢子处理细胞后可以检测出凋亡细胞。 Western blot方法检测到凋亡抑制蛋白bcl－2、bcl－xl随灵芝孢子浓度增加而下降，而促凋亡蛋白bax及P27、casep具有抑制卵巢癌细胞生长及诱导凋亡的作用，其机制是作用于细胞周期的ase－3逐渐升高。结论灵芝孢子粉可以抑制卵巢细胞生长，使细胞生长阻滞于G1期，并可以通过上调bax、P27，下调bcl－2并活化casepase－3而发挥其作用。