摘 要: 目的 探讨Wnt/β-catenin通路在镍精炼烟尘诱导人支气管上皮(Beas-2B)细胞产生Warburg效应过程中的作用。方法 采用不同浓度镍精炼烟尘(0、12.5、25.0、50.0 μg/ml)处理Beas-2B细胞24 h,通过CCK-8法检测镍精炼烟尘的细胞毒性,使用乳酸试剂盒检测乳酸分泌量,使用免疫印迹法(Western blot)检测Wnt/β-catenin信号通路蛋白β-catenin和Warburg效应相关蛋白丙酮酸脱氢酶激酶1(pyruvate dehydrogenase kinase 1,PDK1)、单羧酸乳酸转运体1(monocarborxylat transporter 1,MCT1)和乳酸脱氢酶激酶A(lactate dehydrogenase A,LDHA)的表达水平。采用通路抑制剂XAV-939处理Beas-2B后以25.0 μg/ml镍精炼烟尘染毒24 h,检测Warburg效应相关蛋白PDK1、MCT1和LDHA的表达水平。结果 与对照组相比,镍精炼烟尘染毒组Beas-2B细胞的相对存活率明显降低,乳酸生成量随着镍精炼烟尘暴露浓度增加而升高(P<0.05),β-catenin及Warburg效应相关蛋白PDK1、MCT1和LDHA的表达水平均见增高且呈剂量依赖关系;Wnt/β-catenin通路抑制剂XAV-939可以抑制Warburg效应相关蛋白的表达(P<0.05)。结论 镍精炼烟尘可能是通过激活Wnt/β-catenin信号通路诱导Beas-2B细胞产生Warburg效应,从而将正常的线粒体氧化磷酸化转变为有氧糖酵解的能量代谢方式,并可能诱导癌症的发生。 |
关键词: 镍精炼烟尘 Wnt/β-catenin通路 Warburg效应 |
中图分类号: R994.3
文献标识码: A
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基金项目: 国家自然科学基金资助项目(编号:81973010,82273601) |
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Nickel refining fumes may induce Warburg effect via Wnt/β-catenin pathway activation in human bronchial epithelial cells |
GAO Ying,ZHANG Tong,ZHANG Dan,CHEN Yang-yang,YAO Wen-xue,YANG Shi-kuan,LI Li-na,WU Yong-hui
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School of Public Health,Harbin Medical University,Harbin,Heilongjiang 150081,China
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Abstract: Objective To explore the role of Wnt/β-catenin pathway played in Ni-refining fumes induced Warburg effect in human bronchial epithelial cells(Beas-2B). Methods Beas-2B cells were treated with different concentrations of Ni-refining fumes(0,12.5,25.0,50.0 μg/ml)for 24 h,then the cytotoxicity was detected by CCK-8 assay,the amount of lactic acid secretion was detected using lactic acid kits,the expressions of the signal pathway protein β-catenin of Wnt/β-catenin pathway and the Warburg effect related proteins PDK1、MCT1 and LDHA were detected by Western Blot;additionally,after treating with the pathway inhibitor XAV-939,the Beas-2B cells were exposed to 25.0 μg/ml of nickel refining fume for 24 h,detect the expression levels of PDK1,MCT1 and LDHA again. Results The results showed that compared with the control group,the relative survival rate of Beas-2B cells in Ni-refining fume exposed group was significantly decreased and the lactic acid production amount was rised with the increase of exposure concentration of nickel refining fumes(P<0.05);meanwhile,the expression levels of Warburg effect related proteins such as β-catenin、PDK1、MCT1 and LDHA were all significantly increased in a dose-dependent manner(P<0.05),and the Wnt/β-catenin pathway inhibitor XAV-939 can inhibit the expression of Warburg effect related proteins(P<0.05). Conclusion The results suggested that Ni-refining fumes may induce Warburg effect produced by Beas-2B cells via activation of Wnt/β-catenin pathway,thus,the normal mitochondrial oxidative phosphorylation process will be transformed into the energy metabolism mode of aerobic glycolysis,which maybe eventually lead to cancer. |
Keywords: nickel refining fumes Wnt/β-catenin pathway Warburg effect |