miR-34a调控肺癌EGFR耐药细胞株中MET信号通路探讨

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。

miR-34a Regulating MET Signaling Pathway in EGFR Resistant Lung Cancer Cell Lines

Purpose] To study the miR-34a regulating MET signaling pathway in EGFR resistant lung cancer cell lines,and to investigate the mechanisms of acquired resistance of lung cancer. Methods] On the basis of HCC827 cells（epidermal growth factor receptor gene exon 19 deletions in lung adenocarcinoma cell lines）,the gefitinib resistant cell lines were cultured. the expression of MET and miR-34a in the drug-resistant cell lines were detected by RT-PCR method. The downstream target of miR-34a were predicted by bioinformatics software such as TargetScan. Whether miR-34a could regulate the expression of MET in the intracellular was verified. Results] HCC827/GR cell lines resistant to gefitinib approximately 100-fold compared with HCC827 cells. Low expression of miR-34a and high expression of MET in the resistant cell lines was found,while high expression of miR-34a and low expression of MET in the HCC827 cells. TargetScan bioinformatics software indicated MET is one of downstream target of miR-34a. The fluorescence was decreased when the vectors carried fluorescent reporter group and MET 3′UTR and miR-34 were co-transfected into cells.Conclusion] MiR-34a involves in EGFR-TKI acquired resistance.It might regulate target genes MET.