目的探讨两种环境内分泌干扰物双酚A(bisphenolA,BPA)与玉米赤霉烯酮(zearalenone,ZEA)对人神经母细胞瘤SK—N—SH细胞细胞周期素CyclinDl、CDK4、p21表达的影响。方法SK—N.SH细胞经培养扩增后分为7组:组1,无干预(对照组);组2.加17β-雌二醇(Ez);组3,加BPA,组4.加ZEA;组5,同时加E2和ICI182,780;组6,同时加BPA和ICI182.780;组7,同时加ZEA和ICI182,780。在72h用免疫细胞化学染色及Westernblot分析法检测细胞周期蛋白CyclinDl、CDK4、p21的蛋白表达。结果72h时免疫组化和Westernblot检测发现,组2、3、4中CyclinDl、CDK4的表达明显高于对照组和5、6、7组(P〈0.05),p21在各组之间差异无统计学意义(P〉0.05)。结论BPA与ZEA可能通过调节细胞周期蛋白而影响SK—N—SH细胞的周期变化,从而促进其增殖。
[Abstract] Objective To investigate the effect of environmental endocrine disruptors, bisphenol A (BPA) and zearalenone (ZEA), on the expression of CyclinD1, CDK4 and p21 in SK-N-SH human neuroblastoma (NB) cells in vitro. Methods NB cells were cultured in estrogen free improved minimum essential medium and divided into 7 groups based on different treatments: group 1, no treatment; group 2, treated with 17β-estradiol (E2) ; group 3, treated with BPA; group 4, treated with zearalenone (ZEA) ; group 5, treated with both E2 and ICI 182,7801 group 6, treated with both BPA and ICI 182,780; group 7, treated with both ZEA and ICI 182,780. Western blot analysis and immunol cytochemistry were employed to monitor the expression of CyclinD1, CDK4 and p21. Results At 72 hours, the expression of CyclinD1 and CDK4 was abundant in 2, 3 and 4 group, but was little in control and 5, 6, 7 group (P〈0.05). There was no difference on expression of p21 among the groups (P〈0.05). Conclusions Disrupting cell cycle may be involved in the process of growth of SK-N-SH cells, which is directed by BPA and ZEA.