Wang Jiun-Long, Lan Ying-Wei, Tsai Yi-Ting, Chen Ying-Cheng, Staniczek Theresa, Tsou Yung-An, Yen Chih-Ching, Chen Chuan-Mu
Ph.D. Program in Translational Medicine, Department of Life Sciences, National Chung Hsing University, Taichung, Taiwan.
Division of Chest Medicine, Department of Internal Medicine, Taichung Veterans General Hospital, Taichung, Taiwan.
Front Cell Dev Biol. 2021 Jun 21;9:688062. doi: 10.3389/fcell.2021.688062. eCollection 2021.
Lung cancer is heterogeneous and challenging to cope with once it has progressed. Chemotherapy is the first step once no active driver mutation has been discovered. Non-antitumor drugs have been found to be beneficial when used as adjuvants to chemotherapy. In this study, the additive effect and mechanism of metformin combined with pemetrexed in non-small-cell lung cancer (NSCLC) cells were elucidated. Three NSCLC cell lines, A549, H1975, and HCC827, were used to analyze tumor cell proliferation, colony formation and the cell cycle when exposed to metformin alone, pemetrexed alone or their combination. We found that combination treatment in three cell lines exerted antiproliferative effects through cell cycle arrest in the S phase. An chicken chorioallantoic membrane (CAM) assay was used to examine the antiangiogenic effect of metformin combined with pemetrexed on vascular structure formation. We further created an A549 orthotopic xenograft model with an imaging system (IVIS) and explored the associated indicators involved in the tumorigenic process. The results showed that the combination of metformin and pemetrexed exhibited an antiproliferative effect in reducing cell viability and colony formation, the downregulation of cyclin D1 and A2 and the upregulation of CDKN1B, which are involved in the G1/S phase. For antiangiogenic effects, the combination therapy inhibited the vascular structure, as proven by the CAM assay. We elucidated that combination therapy could target VEGFA and Endoglin by RT-qPCR, ELISA and histopathological findings in an A549 orthotopic NSCLC xenograft model. Our research demonstrated the additive antiproliferative and antiangiogenic effects of the combination of metformin with pemetrexed in NSCLC and could be applied to clinical lung cancer therapy.
肺癌具有异质性，一旦病情进展，治疗颇具挑战性。若未发现有活性的驱动基因突变，化疗是首选治疗手段。已发现非抗肿瘤药物作为化疗辅助药物使用时有益。在本研究中，阐明了二甲双胍联合培美曲塞在非小细胞肺癌（NSCLC）细胞中的附加效应及机制。使用三种NSCLC细胞系A549、H1975和HCC827，分析单独暴露于二甲双胍、培美曲塞或二者联合时肿瘤细胞的增殖、集落形成及细胞周期。我们发现，三种细胞系中的联合治疗通过使细胞周期停滞于S期发挥抗增殖作用。采用鸡胚绒毛尿囊膜（CAM）试验检测二甲双胍联合培美曲塞对血管结构形成的抗血管生成作用。我们进一步利用成像系统（IVIS）建立了A549原位异种移植模型，并探索了肿瘤发生过程中涉及的相关指标。结果显示，二甲双胍与培美曲塞联合使用在降低细胞活力和集落形成方面具有抗增殖作用，可下调参与G1/S期的细胞周期蛋白D1和A2，并上调CDKN1B。对于抗血管生成作用，联合治疗抑制了血管结构，这在CAM试验中得到证实。我们通过RT-qPCR、ELISA以及A549原位NSCLC异种移植模型中的组织病理学发现阐明，联合治疗可靶向VEGFA和内皮糖蛋白。我们的研究证明了二甲双胍与培美曲塞联合在NSCLC中具有附加的抗增殖和抗血管生成作用，可应用于临床肺癌治疗。
