慢病毒靶向溴样结构域蛋白4通过抑制SHH通路对非小细胞肺癌细胞A549能量代谢水平的影响

Effect of lenti-siRNA against BRD4 on energy metabolism in non-small cell lung cancer A549 cells by inhibiting SHH pathway

目的:探究溴样结构域蛋白4(BRD4)siRNA通过SHH通路对非小细胞肺癌细胞A549能量水平的影响。方法:RT-qPCR分析非小细胞肺癌细胞A549、HLF-1细胞内BRD4表达水平;将BRD4 siRNA转入A549细胞内,细胞计数试剂盒-8(CCK-8)和细胞克隆形成实验检测BRD4 siRNA对A549生长增殖的影响;2-[N-(7-硝基苯并-2-氧-1,3-二唑-4-基)氨基]-2-脱氧葡萄糖法(2-NBDG)法和乳酸检测试剂盒检测BRD4 siRNA对A549细胞葡萄糖摄取率和乳酸生成量的影响;RT-qPCR和Western blot检测BRD4 siRNA对基因SHH、胶质瘤相关癌基因同源物1(GLI1)的mRNA和蛋白表达水平的影响。结果:BRD4基因在A549细胞中上调表达(P<0.05);BRD4 siRNA对A549细胞的增殖、克隆形成、葡萄糖摄入和乳酸生成具有抑制作用(P<0.05),且能够促进SHH和GLI1基因的mRNA和蛋白表达显著下调(均P<0.05)。结论:慢病毒靶向BRD4基因能够通过抑制SHH通路降低非小细胞肺癌细胞A549的能量代谢水平。

Objective: To investigate the effect of lenti-siRNA against BRD4 on energy metabolism in non-small cell lung cancer A549 cells by SHH pathway. Methods: The expression of BRD4 in A549 and HLF-1 cells was detected by RT-qPCR. After BRD4 siRNA was transferred into A549 cells, the effects of BRD4 siRNA on the growth and proliferation of A549 cells were detected by CCK8 and cell cloning assays. The effects of BRD4 siRNA on the glucose uptake and lactic acid production of A549 cells were detected by 2-NBDG and lactic acid test kit. The expression levels of SHH, GLI1 mRNA and protein were detected by RT-qPCR and Western blotting respectively. Results: The expression of BRD4 mRNA was up-regulated in A549 cells(P<0.05), and BRD4 siRNA inhibited the proliferation, cloning, glucose intake and lactic acid production of A549 cells(P<0.05), and promoted the significant down-regulation of mRNA and protein expression of SHH and GLI1 genes(P<0.05). Conclusion: The lenti-siRNA against BRD4 could reduce the level of energy metabolism in non-small cell lung cancer A549 cells by inhibiting the SHH pathway.