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Circ-MAN1A2 Contributes to the Acquired Resistance of Gefitinib by Binding to miR-409-3p to Induce TWIST1 Expression in Non-small-cell Lung Cancer

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Abstract

Gefitinib has been widely used as a first-generation epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor for non-small-cell lung cancer (NSCLC) patients with EGFR mutation. In this study, we explored the key molecules responsible for acquired Gefitinib resistance in NSCLC cells. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was performed to analyze the Gefitinib resistance and proliferation ability of NSCLC cells. Cell proliferation was also assessed by colony formation assay. Flow cytometry was performed to analyze cell apoptosis. Wound healing assay and transwell invasion assay were performed to the migration and invasion abilities of NSCLC cells, respectively. The target relationship between microRNA-409-3p (miR-409-3p) and circular RNA mannosidase alpha class 1A member 2 (circ-MAN1A2) or twist family bHLH transcription factor 1 (TWIST1) was verified by dual-luciferase reporter assay and RNA immunoprecipitation assay. Circ-MAN1A2 level was markedly elevated in Gefitinib-resistant NSCLC cell lines and tissues. Circ-MAN1A2 interference sensitized Gefitinib-resistant NSCLC cells to Gefitinib. Furthermore, circ-MAN1A2 interference suppressed the proliferation, migration and invasion and promoted the apoptosis of Gefitinib-resistant NSCLC cells. Circ-MAN1A2 over-expression negatively regulated miR-409-3p level by directly binding to it. miR-409-3p silencing partly counteracted circ-MAN1A2 silencing-mediated anti-tumor effects in Gefitinib-resistant NSCLC cells. TWIST1 was a target of miR-409-3p, and miR-409-3p overexpression-induced anti-tumor effects in Gefitinib-resistant NSCLC cells were partly reversed by TWIST1 overexpression. Circ-MAN1A2 silencing aggravated Gefitinib-mediated inhibition of tumor growth in vivo. In conclusion, circ-MAN1A2 facilitated the acquired resistance of Gefitinib and other malignant behaviors of NSCLC cells through mediating miR-409-3p/TWIST1 axis.

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Authors and Affiliations

  1. Department of Oncology, Jingmen No.1 People’s Hospital, Jingmen, Hubei, 448000, China

    Yun Li & Rong Luo

  2. Department of Medical School, Jingchu University of Technology, Jingmen, Hubei, 448000, China

    Jinping Liu

  3. Department of General Practice, Jingmen No.1 People’s Hospital, Jingmen, Hubei, 448000, China

    Yong You

  4. Department of Oncology 1, Jingmen No.2 People’s Hospital, Jingmen, Hubei, 448000, China

    Guiming Chen

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  1. Yun Li
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  2. Jinping Liu
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  3. Rong Luo
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Correspondence to Guiming Chen.

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The authors declare no conflict of interest. The design of this protocol follows the tenets of the Declaration of Helsinki, approved by the Ethics Committee of Jingmen No.1 People’s Hospital. (No. JM2019071835).

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Li, Y., Liu, J., Luo, R. et al. Circ-MAN1A2 Contributes to the Acquired Resistance of Gefitinib by Binding to miR-409-3p to Induce TWIST1 Expression in Non-small-cell Lung Cancer. Biotechnol Bioproc E 27, 556–571 (2022). https://doi.org/10.1007/s12257-021-0190-2

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