Abstract

It has been reported that clustered miRNAs can be transcribed coordinately and exhibit similar functions by regulating the same targets. miR-1/133a and miR-206/133b are well-characterized miRNA clusters. However, the effect of these clusters on EGFR-TKI resistance is not clear. In this study, we demonstrated that lentivirus-mediated HGF overexpression was able to induce gefitinib resistance in non-small cell lung cancers with EGFR sensitive mutations. miR-1/133a and miR-206/133b clusters could overcome HGF induced gefitinib resistance. Furthermore, the clusters were more effective than individual miRNA. Transcriptome RNA sequencing and bioinformatics analysis revealed that multiple pathways, including ‘EGFR tyrosine kinase inhibitor resistance’ pathway, were involved in anti-resistance mechanisms of miR-1/133a and miR-206/133b clusters. Western blotting results confirmed the inhibitory effect of miRNA clusters on MET expression and downstream pathway activation. In conclusion, miR-1/133a and miR-206/133b clusters are able to exhibit the synergetic effect on overcoming HGF-induced gefitinib resistance in NSCLC and the mechanisms are through targeting multiple genes related to gefitinib resistance.

摘要

据报道，聚集的miRNA可以通过调节相同的靶标来协调转录并表现出相似的功能。miR-1/133a 和 miR-206/133b 是充分表征的 miRNA 簇。然而，这些簇对 EGFR-TKI 耐药性的影响尚不清楚。在这项研究中，我们证明了慢病毒介导的 HGF 过表达能够在具有 EGFR 敏感突变的非小细胞肺癌中诱导吉非替尼耐药。miR-1/133a 和 miR-206/133b 簇可以克服 HGF 诱导的吉非替尼耐药性。此外，簇比单个 miRNA 更有效。转录组 RNA 测序和生物信息学分析显示，包括“EGFR 酪氨酸激酶抑制剂抗性”途径在内的多种途径参与了 miR-1/133a 和 miR-206/133b 簇的抗抗性机制。Western印迹结果证实了miRNA簇对MET表达和下游通路激活的抑制作用。总之，miR-1/133a 和 miR-206/133b 簇能够在克服 HGF 诱导的 NSCLC 中的吉非替尼耐药方面表现出协同作用，其机制是通过靶向与吉非替尼耐药相关的多个基因。