阴离子通道CFTR在双酚A致前列腺癌细胞凋亡的机制研究

Study on the Mechanism of Anion Channel CFTR in Bisphenol A-inducedApoptosis of Prostate Cancer Cells

目的:探讨阴离子通道囊性纤维化跨膜传导调节蛋白(CFTR)表达水平与双酚A致前列腺癌细胞系凋亡中的机制。方法:将前列腺癌细胞系PC-3和LNCaP按染毒剂量分为DMSO溶剂对照组、0μM组、低、中、高剂量组,分别暴露于含不同染毒剂量的溶液24 h。用MTT和酶联免疫吸附法(ELISA)检测BPA对细胞的毒性反应;采用酶标仪检测ATP的浓度;ELISA法检测经细胞系中cAMP和CFTR的表达水平;Western-blotting法检测MAPKα、BCl-2和Bax的浓度;Annexin-FITC与PI染色-流式细胞仪定量检测细胞凋亡水平。用单因素方差分析比较组间差异。结果:经不同浓度BPA染毒24 h后,PC-3和LNCaP细胞的细胞存活率随着BPA浓度的增加而降低;cAMP、CFTR和ATP的表达水平降低;Bax蛋白表达增加而Bcl-2蛋白表达减少,AMPKα激酶表达增加,细胞凋亡数目增加。结论:BPA诱导的前列腺癌可能与离子通道CFTR影响ATP合成、细胞凋亡途径有关。

Objective:To explore the mechanism of anion channel cystic fibrosis transmembrane conductance regulator(CFTR)expression and bisphenol A-induced apoptosis in prostate cancer cell lines.Methods:The prostate cancer cell lines PC-3 and LNCaP were divided into DMSO solvent control group,0μM group,low,medium,and high dose groups according to the exposure dose.They were respectively exposed to solutions containing different doses for 24 hours.MTT and ELISA were used to detect the toxicity of BPA on cells.ATP concentration was measured by a microplate reader.ELISA was used to detect the expression levels of cAMP and CFTR in cell lines.Western-blotting was used to detect MAPKαand Bcl-2 and Bax concentrations.Annexin V-FITC staining and flow cytometry was applied to quantitatively determine apoptosis levels.Variation between groups was compared with one-way analysis of variance.Results:After treated with different concentrations of BPA for 24 hours,the cell viability of PC-3 and LNCaP cells was decreased with increasing BPA concentration.The expression levels of cAMP,CFTR,ATP decreased;Bax protein expression increased while Bcl-2 protein expression decreased,AMPKαexpression increased,and the number of apoptosis increased.Conclusion:BPA-induced prostate cancer perhaps is possibly relevant with the anion channel CFTR by affecting ATP synthesis and cell apoptosis.