雷公藤甲素对人卵巢癌A2780细胞抑制作用的机制研究

Mechanism of Triptolide Inhibiting Human Ovarian Cancer A2780 Cells

目的观察雷公藤甲素作用于人卵巢癌A2780细胞后,细胞增殖和凋亡的变化,并探讨其作用前后的分子机制。方法配置不同浓度的雷公藤甲素溶液,将其作用于A2780细胞,采用MTT法检测不同浓度雷公藤甲素作用A2780细胞后,细胞生长的抑制情况,采用荧光定量聚合酶链反应及Western blot检测不同浓度雷公藤甲素作用A2780细胞前后磷脂酰肌醇-3-激酶(PI3K)、蛋白激酶B(Akt)、B细胞淋巴瘤/白血病-2(Bcl-2)和Bcl-2相关X蛋白(Bax)信使RNA(mRNA)及蛋白的表达情况。结果不同浓度雷公藤甲素分别作用于A2780细胞24、48、72 h后,MTT结果显示,随着雷公藤甲素浓度的升高,作用时间的延长,抑制A2780细胞生长作用逐渐增强。10 nmol/L组的Bcl-2 mRNA表达低于0 nmol/L组[(0.62±0.10)比(1.00±0.21)](P<0.05);20 nmol/L组的Bcl-2 mRNA表达低于0 nmol/L组[(0.37±0.10)比(1.00±0.21)](P<0.01),Bax mRNA表达高于0 nmol/L组[(1.48±0.33)比(1.00±0.13)](P<0.05);40 nmol/L组的PI3K、Akt、Bcl-2 mRNA表达均低于0 nmol/L组[(0.45±0.10)比(1.00±0.56),(0.49±0.41)比(1.00±0.11),(0.28±0.10)比(1.00±0.21)](P<0.05),Bax mRNA表达高于0 nmol/L组[(5.07±0.78)比(1.00±0.13)](P<0.01)。结论雷公藤甲素可抑制人卵巢癌A2780细胞增殖并诱导凋亡,其机制可能与下调PI3K、Akt和Bcl-2 mRNA和蛋白,及上调Bax mRNA和蛋白的表达有关。

Objective To observe the changes of proliferation and apoptosis of human ovarian cancer A2780 cells treated with triptolide,and to explore the molecular mechanism before and after its action.Methods A2780 cells were tested with different concentrations of triptolide.MTT assay was adopted to test the cell proliferation inhibition after treatment with different concentrations of triptolide.The expressions of phosphatidylinositol-3-kinase(PI3K),protein kinase B(Akt),B-cell lymphoma/leukemia-2(Bcl-2)and Bcl-2 related X protein(Bax)messenger RNA(mRNA)and protein were detected by fluorescence quantitative polymerase chain reaction and Western blot.Results After different concentrations of triptolide acting on A2780 cells for 24,48 and 72 h,MTT results showed that with the increase of triptolide concentration and the extension of action time,the inhibitory effect on the growth of A2780 cells gradually increased.The expression of Bcl-2 mRNA in 10 nmol/L group was lower than that in 0 nmol/L group[(0.62±0.10)vs(1.00±0.21)](P<0.05);the expression of Bcl-2 mRNA in 20 nmol/L group was lower than that in 0 nmol/L group[(0.37±0.10)vs(1.00±0.21)](P<0.01),and the expression of Bax mRNA in 20 nmol/L group was higher than that in 0 nmol/L group[(1.48±0.33)vs(1.00±0.13)](P<0.05);the mRNA expressions of PI3K,Akt and Bcl-2 in 40 nmol/L group were lower than those in 0 nmol/L group[(0.45±0.10)vs(1.00±0.56),(0.49±0.41)vs(1.00±0.11),(0.28±0.10)vs(1.00±0.21)](P<0.05),and the Bax mRNA expression was higher than that in 0 nmol/L group[(5.07±0.78)vs(1.00±0.13)](P<0.01).Conclusion Tripto-lide can inhibit the proliferation and induce the apoptosis of human ovarian cancer A2780 cells,and its mechanism may be connected with down-regulating the mRNA and protein expression of PI3K,Akt and Bcl-2 and up-regulating the mRNA and protein expression of Bax.