3,3-二吲哚甲烷在体外和体内对卵巢癌SKOV3细胞增殖的影响

Effect of 3,3’-diindolylmethane on the proliferation of human ovarian cancer cell SKOV3 in vitro and in vivo

目的:探讨3,3-二吲哚甲烷(3,3-diindolylmethane,DIM)对卵巢癌细胞SKOV3体外增殖以和体内成瘤能力的影响,以及可能的作用机制。方法:采用不同浓度的DIM处理SKOV3细胞后,观察DIM对SKOV3细胞形态的影响;进一步用Hoechst33342染色后,观察DIM对SKOV3细胞凋亡的影响;随后采用CCK-8法和平板克隆形成实验检测DIM对SKOV3细胞增殖能力的影响。皮下接种SKOV3细胞构建BALB/c裸鼠移植瘤模型,通过灌胃给药途径观测DIM对SKOV3细胞移植瘤生长的影响;采用免疫组织化学法检测DIM对肿瘤组织中AKT、磷酸化AKT(phospho-AKT,p-AKT)及Bcl-2和caspase3蛋白表达水平的影响。结果:DIM能明显改变SKOV3细胞的形态,并诱导其凋亡。DIM能明显抑制SKOV3细胞的增殖能力和克隆形成能力(P<0.05),并呈一定的浓度依赖性。体内研究显示,DIM可显著抑制小鼠SKOV3细胞移植瘤的生长能力(P<0.05);免疫组织化学法检测结果可见,DIM可明显下调移植瘤中p-AKT和Bcl-2蛋白的表达水平,而上调caspase3的表达水平(P<0.05),但对AKT总蛋白表达无明显影响。结论:DIM可有效抑制卵巢癌细胞的体外增殖能力及体内成瘤能力,其作用机制可能与抑制AKT的活化以及Bcl-2表达,诱导caspase3表达有关。

Objective:To investigate the effects of 3,3’-diindolylmethane(DIM)on the proliferation and tumorigenicity of ovarian cancer cell SKOV3 in vitro and in vivo and the underlying mechanism.Methods:The effect of DIM on the morphology of SKOV3 cells was observed after treatment with varied concentrations of DIM,and the effect of DIM on the apoptosis of SKOV3 cells was further assessed after Hoechst33342 staining.The effect of DIM on the proliferation of SKOV3 cells was evaluated by CCK-8 assay and colony formation assay.The tumor xenograft model was established by subcutaneous injection of SKOV3 cells into BALB/c nude mice.The effect of DIM on the growth of the SKOV3 xenograft tumor was observed after drug administration through oral gavage.Immunohistochemical analysis was used to examine the effect of DIM on the expression levels of AKT,p-AKT(phospho-AKT),Bcl-2 and caspase3 in xenograft tumor tissues.Results:DIM could obviously change the morphology and induce the apoptosis of SKOV3 cells.DIM could significantly inhibit the proliferation and colony formation of SKOV3 cells in a dose dependent manner(P<0.05).The in vivo experimental results showed that DIM could significantly suppress the growth of SKOV3 xenograft tumors(P<0.05).Immunohistochemical analysis results showed that DIM could obviously down-regulate the expression levels of p-AKT and Bcl-2 while upregulate the expression level of caspase3(P<0.05)without affecting the expression level of total AKT protein.Conclusion:DIM can effectively inhibit the proliferation of ovarian cancer cells in vitro and the tumorigenicity of ovarian cancer cells in vivo.The mechanism may be associated with the inhibition of AKT activation and Bcl-2 expression,and the indu ction of caspase3 expression.