碳酸酐酶1在非小细胞肺癌早期发病中作用机制的研究

Mechanism of carbonic anhydrase 1 in the early stage of non-small cell lung cancer

目的:探讨碳酸酐酶1(carbonic anhydrase 1,CA1)在非小细胞肺癌(non-small cell lung cancer,NSCLC)早期诊断的价值及其在肺癌早期发病中的作用机制。方法:应用酶联免疫吸附实验检测临床NSCLC患者血清中CA1的含量;应用RNA干扰及质粒扩增的方法分别对A549、H520、H1299细胞进行处理,构建敲低和过表达CA1的细胞模型,并应用CCK-8法检测不同处理组中细胞的增殖情况;应用实时定量多聚酶链式反应(polymerase chain reaction,PCR)和Western blot方法检测WNT/β-catenin信号通路中关键信号分子Wnt3a和β-catenin mRNA和蛋白质的表达水平。结果:CA1在早期NSCLC患者的血清中表达水平显著高于健康人群。通过sh-RNA抑制CA1表达后,A549、H520和H1299细胞系的增殖速率显著降低,而过表达CA1后,细胞增殖速率明显提高。同时,CA1能够有效调节WNT/β-catenin信号通路中Wnt3a及β-catenin的表达水平。结论:CA1通过Wnt/β-catenin信号通路影响NSCLC的增殖过程;CA1有望成为NSCLC早期诊断的新型生物分子标记物。

Objective:To investigate the diagnostic value and mechanism of carbonic anhydrase 1(CA1)in the early stage of non-small cell lung cancer(NSCLC).Methods:Serum CA1 levels in patients with early stage NSCLC were determined by ELISA.In vitro(in A549,H520,and H1299 cells),we used RNA interference to knock down the expression of CA1 and used plasmid transfection to establish a CA1 over-expression model.A CCK-8 assay was used to detect cell proliferation in different treatment groups.Real-time quantitative polymerase chain reaction(PCR)and Western blot were used to detect the mRNA and protein expression levels of Wnt3a andβ-catenin,which are the key components in the Wnt/β-catenin signaling pathway.Results:The expression of CA1 in the serum of patients with early NSCLC was significantly higher than that in healthy individuals.After knocking down the expression of CA1 by sh-RNA,the proliferation rates of A549,H520,and H1299 were significantly reduced,while after overexpression of CA1,the cell proliferation rate increased.CA1 can also effectively regulate the expression levels of Wnt3a andβ-catenin in the Wnt/β-catenin signaling pathway.Conclusions:CA1 may affect the proliferation of NSCLC through the Wnt/β-catenin signaling pathway,especially by Wnt3a andβ-catenin;CA1 is expected to become a new biomolecular marker for the early diagnosis of NSCLC.