【摘要】 目的研究前B细胞克隆增强因子(PBEF)对缺氧/复氧(H/R)处理的非小细胞肺癌(NSCLC)细胞生长及水通道蛋白1(AQP1)和钠通道蛋白α(ENaCα)表达的影响。方法将NSCLC A549细胞分为4组:对照组(C组)、H/R处理模型组(M组)、H/R处理+pCAGGS空载组(pCAGGS NC组)、H/R处理+pCAGGS PBEF组(pCAGGS PBEF组);通过构建pCAGGS PBEF质粒并转染到细胞中建立H/R A549细胞培养实验模型。采用qPCR和Western blot(WB)检测PBEF的表达;采用WB检测AQP1和ENaCα的表达;采用流式细胞术检测细胞凋亡。结果 M组细胞凋亡率显著高于C组(P<0.05),pCAGGS PBEF组细胞凋亡率显著高于pCAGGS NC组(P<0.05),M组和pCAGGS NC组细胞凋亡率比较差异无统计学意义(P>0.05)。pCAGGS PBEF组细胞中AQP1和ENaCα的蛋白表达水平显着低于pCAGGS NC组(P<0.05),而C组、M组、pCAGGS NC组3组细胞中AQP1和ENaCα的蛋白表达水平比较差异均无统计学意义(P>0.05)。结论 PBEF过表达促进了H/R处理的NSCLC A549细胞凋亡,并抑制AQP1和ENaCα蛋白的表达,提示PBEF在低氧环境中可能具有辅助治疗作用。更多还原

【Abstract】 Objective To investigate the effects of pre-B cell colony-enhancing factor(PBEF) on growth of hypoxia/reoxygenation(H/R)-treated human non-small cell lung carcinoma(NSCLC) cells and expression of aquaporin-1(AQP1) and epithelial sodium channel α-subunit(ENaCα).Methods NSCLC A549 cells were divided into four groups:control group(C group),H/R-treated model group(M group),H/R treatment+empty pCAGGS vector group(pCAGGS NC group),and H/R treatment+pCAGGS PBEF group(pCAGGS PBEF group).PCAGGS PBEF plasmids were constructed and transfected into cells to establish the H/R model,and levels of PBEF expression were detected by qPCR and Western blot(WB).The expression AQP1 and ENaCα was determined by WB,and cell apoptosis was detected by flow cytometry.Results The apoptosis rate in M group was higher than that in C group,and that in pCAGGS PBEF group was higher than that in pCAGGS NC group(P<0.05).The difference in apoptosis rate was not significant between M group and pCAGGS NC group(P>0.05).The expression of AQP1 and ENaCα in pCAGGS PBEF group was lower than that in pCAGGS NC group(P<0.05).There were no significant differences in AQP1 and ENaCα expression among C,M and pCAGGS NC groups(P>0.05).Conclusion PBEF overexpression can promote apoptosis and inhibit AQP1 and ENaCα expression in H/R-treated NSCLC A549 cells,suggesting that PBEF may play an adjuvant therapeutic role in hypoxic environment.更多还原