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[目地]研究肺癌耐药细胞株HCC827/GR中MET信号通路的调控,明确肺癌获得性耐药的分子机制。[方法]使用HCC827细胞[epidermal growth factor receptor(EGFR)基因19外显子缺少的肺腺癌细胞株],在此细胞的基础上培养吉非替尼耐药细胞株。检测耐药细胞株中MET的表达,并使用RT-PCR的方法检测miR-34a的表达;使用TargetScan等生物信息学软件预测miR-34a的下游靶点,并在细胞内验证miR-34a是否可以调控MET的表达。[结果]与HCC827细胞相比,HCC827/GR细胞株对吉非替尼明显耐药。在耐药HCC827/GR细胞株中,miR-34a低表达,MET高表达,而在HCC827细胞株中,miR-34a高表达,MET低表达。Target Scan生物信息学软件提示,MET是miR-34a的下游靶点之一。将携带荧光报告基团和MET 3′UTR的载体与miR-34a共转染入细胞后荧光度下降。[结论]miR-34a可能通过调控靶基因MET而参与EGFR-TKI的获得性耐药。
[Objective] To study the regulation of MET signaling pathway in drug-resistant lung cancer cell line HCC827 / GR and elucidate the molecular mechanism of acquired drug resistance in lung cancer. [Method] The lung adenocarcinoma cell lines lacking the exon 19 of epidermal growth factor receptor (EGFR) gene were used in this study. Gefitinib cell lines were cultured on the basis of this cell line. We detected the expression of MET in drug-resistant cell lines and detected the expression of miR-34a by RT-PCR. The bioinformatics software such as TargetScan was used to predict the downstream targets of miR-34a and verify whether miR-34a Regulates MET expression. [Results] HCC827 / GR cell lines were significantly resistant to gefitinib compared with HCC827 cells. In the resistant HCC827 / GR cell line, miR-34a is low-expressed and MET is highly expressed, while in HCC827 cell line miR-34a is highly expressed and MET is low-expressed. Target Scan bioinformatics software suggests that MET is one of the downstream targets of miR-34a. Fluorescence decreased after cotransfection of vector carrying miR-34a and reporter carrying the reporter and MET 3’UTR into cells. [Conclusion] miR-34a may be involved in acquired resistance of EGFR-TKI by regulating target gene MET.