乳酸通过GPR81介导的Nrf2/LDHA通路促进非小细胞肺癌H1299细胞的迁移

Lactic acid promotes migration of non-small cell lung cancer H1299 cells via GPR81-mediated Nrf2/LDHA pathway

目的:探讨乳酸诱导GPR81、Nrf2、LDHA等相关蛋白表达对非小细胞肺癌H1299细胞迁移的影响。方法:体外培养H1299,加入乳酸、siGPR81、PI3K-AKT通路抑制剂LY294002和Nrf2 siRNA处理,采用Western blot检测乳酸对GPR81、Nrf2和LDHA的调控作用;构建不同片段LDHA启动子质粒,通过双荧光素酶报告基因实验检测Nrf2对LDHA转录水平的影响;采用Transwell实验检测乳酸对H1299细胞迁移的影响,细胞划痕实验检测LDHA对H1299细胞迁移的影响。结果:乳酸诱导GPR81、Nrf2和LDHA蛋白表达并促进迁移;GPR81通过PI3K-AKT通路正调控Nrf2;Nrf2在转录和翻译水平正调控LDHA的表达;沉默LDHA的表达可以抑制H1299细胞的迁移。结论:乳酸可以通过GPR81介导的Nrf2/LDHA通路促进非小细胞肺癌H1299细胞的迁移。

Objective:To investigate the effects of lactic acid-induced expression of GPR81,Nrf2,LDHA and other related proteins on the migration of non-small cell lung cancer(NSCLC)cell line H1299.Methods:H1299 was cultured and treated with lactic acid,siGPR81,PI3K-AKT pathway inhibitor LY294002 and Nrf2 siRNA,and Western blot was used to detect the regulatory effects of lactic acid on GPR81,Nrf2 and LDHA.Different fragments of LDHA promoter plasmids were constructed to detect the effect of Nrf2 on LDHA transcription through dual luciferase reporter assays.Transwell test was used to detect the effect of lactic acid on the migration of H1299,and cell scratch test was used to detect the effect of LDHA on the migration of H1299.Results:Lactic acid promoted the expression of GPR81,Nrf2 and LDHA proteins and the migration of H1299.GPR81 positively regulates Nrf2 ex⁃pression through PI3K-Akt pathway.Nrf2 positively regulates LDHA expression at the transcription and translation levels.Interference of LDHA expression could inhibit the migration of H1299.Conclusion:Lactic acid promotes the migration of NSCLC cell line H1299 through the Nrf2/LDHA pathway mediated by GPR81.