WT1在非小细胞肺癌中的表达及其与肺癌细胞增殖的关系

The expression of WT1 in non-small cell lung cancer and its relationship with proliferation of lung cancer H1299 cells

目的:探讨人非小细胞肺癌及其相应癌旁组织中WT1基因(Wilms’tumor gene1,WT1)的表达,体外研究其与肺癌细胞增殖的关系。方法:应用实时荧光定量PCR法检测79例非小细胞肺癌组织及其相应癌旁组织中WT1mRNA的表达情况。构建重组质粒pLV-GFP-WT1,并通过脂质体LipofectAMINE2000将其瞬时转染至非小细胞肺癌H1299细胞中,应用蛋白质印迹法检测转染后H1299细胞中WT1蛋白的表达,CCK-8(cell countingkit-8)法检测细胞的增殖情况。结果:非小细胞肺癌组织中WT1mRNA的表达水平高于相应的癌旁组织,差异有统计学意义(P<0.01)。重组质粒pLV-GFP-WT1成功构建,并将其成功转染至H1299细胞中;转染后的H1299细胞中,WT1蛋白的表达水平上调;在pLV-GFP-WT1转染后24、36和48h时,过表达WT1的H1299细胞增殖率高于对照组,差异有统计学意义(P<0.05)。结论:WT1mRNA在非小细胞肺癌组织中的表达水平明显高于相应癌旁组织,高表达WT1基因能促进非小细胞肺癌H1299细胞的增殖,提示WT1在非小细胞肺癌中扮演癌基因的角色。

Objective： To investigate the expression of WT1 （Wilms’ tumor gene 1） in NSCLC （non-small cell lung cancer） tissues and the corresponding para-cancerous tissues, and to explore the relationship between WT1 expression and the proliferation of NSCLC cells in vitro. Methods： The expression levels of WT1 mRNA in 79 NSCLC tissues and the corresponding para-cancerous tissues were detected by RFQ-PCR （real-time fluorescence quantitative-PCR）. Recombination plasmid of pLV-GFP-WT1 was constructed and transfeced into NSCLC H1299 cells by LipofectAMINE 2000. The expression level of WT1 protein in H1299 cells after transfection with pLV-GFP-WT1 was detected by Western blotting, and the proliferation of H1299 cells was detected by CCK-8 （cell counting kit-8）. Results： The expression level of WT1 mRNA in NSCLC tissues was higher than that in the corresponding para-cancerous tissues （P 〈 0.01）. Recombination plasmid of pLV-GFP-WT1 was successfully constructed and transfected into NSCLC H1299 cells. After transfection, the expression level of WT1 protein was up-regulated in H1299 cells. The proliferation rates of H1299 cells 24, 36 and 48 h after transfection with pLV-GFP-WT1 were higher than those of the cells without transfection （P 〈 0.05）. Conclusion： The expression level of WT1 mRNA in NSCLC tissues is higher than that in the corresponding para-cancerous tissues. The overexpression of WT1 can promote the proliferation of NSCLC H1299 cells. It is suggested that WT1 may act as an oncogene in NSCLC.