三氧化二砷对卵巢癌细胞的生长及化疗敏感性的影响

目的:研究三氧化二砷(As2O3)对人卵巢癌COC1细胞顺铂(cDDP)化疗敏感性的影响,As2O3与cDDP联合作用的效应以及As2O3对COC1细胞的生长抑制效应及机制。方法:四甲基偶氮唑蓝(MTT)比色法检测不同浓度As2O3、cDDP作用72h对卵巢癌细胞生长的抑制作用,As2O3对顺铂化疗敏感性的影响及两药联合作用的效应。逆转录聚合酶链反应(RT-PCR)和免疫印迹技术(Western blot)分别检测PIK3CA mRNA和AKT、ERK、MMP-2蛋白表达的变化。结果:单独应用As2O3或cDDP和联合用药时卵巢癌COC1细胞生长均受到抑制,并呈浓度依赖关系,联合用药组抑制率明显高于单独用药组(P<0.05);0.08~0.15μmol/L As2O3无明显细胞毒性,不能提高COC1细胞对顺铂的敏感性。各浓度(1、3、8、16μmol/L)As2O3作用24h可以下调AKT、MMP-2蛋白水平(P<0.05),并呈浓度依赖效应,但对ERK蛋白的表达无显著影响。0.5、1、2、4μmol/L As2O3作用4h,PIK3CA mRNA表达分别降低45.03%、53.05%、61.67%和72.91%(P<0.05)。结论:As2O3抑制COC1细胞增殖具有浓度依赖性,与顺铂联用有相加效应,可能与As2O3下调PIK3CA mRNA、AKT、MMP-2蛋白水平等有关。

The effect of arsenic trioxide on cell growth and influence on the chemosensitivity of human ovarian cancer cells

Objective:To study the influence of arsenic trioxide(As2O3)on the chemosensitivity of human ovarian cancer cell line COC1 to cisplatin and combination effect of As2O3 and cisplatin,and explore cytotoxic mechanism of As2O3 in COC1 cells.Methods:The inhibition effect of different doses of As2O3 and cisplatin for 72hours and influence of As2O3 on cisplatin chemosensitivity as well as combination effect of As2O3 and cisplatin in COC1 cells were assayed with MTT test.The expression of PIK3CA mRNA and AKT,ERK,MMP-2 protein in COC1 cells after being exposed to As2O3 solution of different concentration were determined by RT-PCR and Western blotting respectively.Results:As2O3 and cisplatin used alone or in combination inhibited the growth of COC1 cells in a dose-dependent manner.The inhibition was more obvious in the combination group than in As2O3 group and cisplatin group(P<0.05);As2O3 at 0.08μmol/L to 0.15μmol/L concentrations was not significantly cytotoxic to COC1 cells,and could not improve the sensitivity of COC1 cells to cisplatin.Different concentration(1,3,8,16μmol/L) of As2O3 treatment for 24hours decreased AKT and MMP-2 protein expression in a dose-dependent fashion(P<0.05),but did not alter ERK protein expression.Compared with the control group,different concentration(0.5,1,2,4μmol/L) of As2O3 treatment for 4hours significantly decreased PIK3CA mRNA expression in a dose-dependent fashion(P<0.05).Conclusion:As2O3 can inhibit proliferation of COC1 cells in a concentration-dependent manner and additive effect of As2O3 in combination with cisplatin is observed,which may be related to down-regulation of the expression of PIK3CA,AKT and MMP-2 expression.