METTL7B促进非小细胞肺癌细胞侵袭的机制研究

Study on the mechanism of METTL7B promoting non-small cell lung cancer in cell invasion

目的探讨甲基转移酶样蛋白-7B(methyltransferase-like 7B,METTL7B)对非小细胞肺癌细胞侵袭的机制。方法通过Western blotting法和qPCR检测METTL7B在A549细胞和正常人上皮细胞分别在蛋白和mRNA水平上表达情况;利用慢病毒在A549细胞中过表达METTL7B,通过Western blotting法检测过表达情况,细胞Transwell侵袭实验检测过表达METTL7B对A549细胞的侵袭能力;再利用双向电泳技术对过表达METTL7B的A549细胞中蛋白质组学进行分析,探讨METTL7B促进非小细胞肺癌细胞侵袭的机制。结果RT-qPCR和Western blotting法检测结果均显示METTL7B在A549中的表达明显高于正常的人肺上皮细胞。使用慢病毒能有效地促使METTL7B在A549中过表达,侵袭实验结果显示过表达METTL7B可增强A549细胞的侵袭能力。通过双向电泳技术分析过表达METTL7B细胞中差异蛋白,发现共有12个差异表达蛋白,其中6个上调,6个下调,且通过Western blotting法实验验证发现过表达METTL7B细胞中的SOD1和SAE1表达上调。结论METTL7B可能通过上调SOD1和SAE1的表达,从而增强非小细胞肺癌细胞的侵袭能力。

Objective To study the mechanism of METTL7B on the invasion of non-small cell lung cancer cells.Methods The protein and mRNA levels of METTL7B in A549 cells and normal human epithelial cells(BEAS-2B)were detected by Western blotting and qPCR,respectively;METTL7B was overexpressed in A549 cells by lentivirus,and the overexpression effect was detected by Western blotting;cell transwell invasion assay was used to detect the invasion ability of METTL7B over expressing in A549 cells,then in order to find the mechanism of METTL7B promoting the invasion of non-small cell lung cancer cells,we used the method of two-dimensional electrophoresis to analyze the proteomics in A549 cells over expressing METTL7B to explore the mechanism of MELLT7B promoting the invasion of non-small cell lung concer cells.Results RT-sqPCR and Western blotting showed that the expression of METTL7B in A549 was significantly higher than that in normal human lung epithelial cells.The lentivirus used can effectively overexpress METTL7B in A549.The invasion experiment showed that overexpression of METTL7B can enhance the invasion ability of A549 cells.The differential proteins in overexpressed METTL7B cells were analyzed by two-dimensional electrophoresis.It was found that there were 12 differentially expressed proteins,of which 6 were up-regulated and 6 were down regulated.The expression of(superoxide dismutase,SOD1)and SAE1 in overexpressed METTL7B cells was up-regulated by Western blotting.Conclusion METTL7B can promate the invasive ability of non-small cell lung cancer cells by up-regulating the expressions of SOD1 and SAE1.