Effect of novel epidermal growth factor receptor isoform EGFRvA on migration of human glioma U87MG cells and its possible mechanism / 肿瘤

ABSTRACT

Objective: To investigate the effect of novel epidermal growth factor receptor variant type A (EGFRvA) on migration of human glioma U87MC cells, and to explore its possible mechanism. Methods: The lentiviral expression vector pWPT-GFP [recombined with green fluorescent protein (GFP) gene, as the blank control], pWPT-EGFRwt (recombined with ECFR wild-type gene, as the negative control) and pWPT-EGFRvA (recombined with ECFRvA gene) was respectively co-transfected with packaging vector psPAX2 and pMD2.G into 293T cells to produce lentiviral particles. Subsequently, the lentiviral particles were separately infected into U87 MG cells to establish U87MG GFP, U87MG EGFRwt and U87MG EGFRvA cell lines. The overexpression levels of EGFRwt and EGFRvA in U87MG EGFRwt and U87MG EGFRvA cells were detected by flow cyctometry (FCM) and Western blotting. The migration capability of U87MG cells after transfection was investigated by Transwell migration test. The real-time fluorescent quantitative-PCR (RFQ-PCR) was performed to validate the varied mRNA expression levels of 52 genes which had been screened out in three types of cells by microarray. Then the protein expression levels of 4 tumor metastasis-related genes which had significantly varied mRNA expression levels in RFQ-PCR results were confirmed by Western blotting. Results: After U87 MG cells were stably infected with lentivirus particles, U87 MG EGFRwt and U87MG EGFRvA cells overexpressed EGFRwt and EGFRvA, respectively. The migration capability of U87MG EGFRvA cells was stronger than that of U87MG EGFRwt cells (P < 0.001). The expression levels of FBLN2 and ROBOI mRNAs were higher in U87MG EGFRvA cells than those in U87MG EGFRwt cells (both P < 0.05), whereas the expression levels of CDH13 and SOX2 mRNAs were the opposite (both P < 0.05). U87MG EGFRvA cells had a significantly lower expression levels of CDH1 3 and SOX2 proteins which were related to tumor metastasis (both P < 0.05). Conclusion: Overexpression of EGFRvA can promote the migration of U87MG cells through regulating the expressions of FBLN2, ROBO1, CDH13 and SOX2 genes in tumor cells.