Xiangfeng Jin1, Yi Yu1, Qiang Zou2, Mingzhao Wang1, Yaojie Cui1, Jing Xie1, Zizong Wang1. 1. Department of Thoracic Surgery, The Affiliated Hospital of Qingdao University, Qingdao, China. 2. Department of Blood Transfusion, Qingdao Haici Hospital Affiliated to Qingdao University, Qingdao, China.
Abstract
BACKGROUND: A growing number of microRNAs have been proved to play significant roles in limiting tumor growth and the epithelial-mesenchymal transition (EMT) process of nonsmall cell lung cancer (NSCLC). Present work aims to study the function of microRNA (miR)-105 in EMT of NSCLC cells, which is unrevealed yet. METHODS: Two NSCLC cell lines A549 and Calu-3 were transfected with miR-105 mimic, inhibitor, or scrambled control. And then the effects of miR-105 were evaluated by performing trypan blue staining, transwell assay, ANNEXIN-FITC/propidium iodide (PI) double staining and Western blot analysis. The expression levels of myeloid cell leukemia-1 (Mcl-1) after transfection were tested by real-time quantitative polymerase chain reaction and Western blot analysis. Whether Mcl-1 was a downstream effector of miR-105, and the involvement of mammalian target of Rapamycin (mTOR) and p38 mitogen-activated protein kinase (p38MAPK) signaling pathways were assessed. RESULTS: The overexpression of miR-105 significantly increased the viability and migration of A549 and Calu-3, but had no impacts on cell apoptosis. Meanwhile, E-cadherin was remarkably downregulated, and N-cadherin, Vimentin, ZEB1, and Snail were upregulated by miR-105 overexpression. Mcl-1 was positively regulated by miR-105, and the effects of miR-105 overexpression on A549 and Calu-3 cells viability, migration and EMT were all flattened by Mcl-1 silence. Both mTOR and p38MAPK pathways were activated in miR-105-overexpressing and Mcl-1-overexpressing cells. Besides, inhibition of mTOR and p38MAPK pathways by using Rapamycin and VX-702 abolished the regulatory effects of Mcl-1 on EMT. CONCLUSION: Our study underlines the importance of miR-105 in modulating NSCLC cells EMT. miR-105 promoted the EMT of NSCLC cells possibly via upregulation of Mcl-1 and thereby activation of mTOR and p38MAPK signaling.
BACKGROUND: A growing number of microRNAs have been proved to play significant roles in limiting tumor growth and the epithelial-mesenchymal transition (EMT) process of nonsmall cell lung cancer (NSCLC). Present work aims to study the function of microRNA (miR)-105 in EMT of NSCLC cells, which is unrevealed yet. METHODS: Two NSCLC cell lines A549 and Calu-3 were transfected with miR-105 mimic, inhibitor, or scrambled control. And then the effects of miR-105 were evaluated by performing trypan blue staining, transwell assay, ANNEXIN-FITC/propidium iodide (PI) double staining and Western blot analysis. The expression levels of myeloid cell leukemia-1 (Mcl-1) after transfection were tested by real-time quantitative polymerase chain reaction and Western blot analysis. Whether Mcl-1 was a downstream effector of miR-105, and the involvement of mammalian target of Rapamycin (mTOR) and p38 mitogen-activated protein kinase (p38MAPK) signaling pathways were assessed. RESULTS: The overexpression of miR-105 significantly increased the viability and migration of A549 and Calu-3, but had no impacts on cell apoptosis. Meanwhile, E-cadherin was remarkably downregulated, and N-cadherin, Vimentin, ZEB1, and Snail were upregulated by miR-105 overexpression. Mcl-1 was positively regulated by miR-105, and the effects of miR-105 overexpression on A549 and Calu-3 cells viability, migration and EMT were all flattened by Mcl-1 silence. Both mTOR and p38MAPK pathways were activated in miR-105-overexpressing and Mcl-1-overexpressing cells. Besides, inhibition of mTOR and p38MAPK pathways by using Rapamycin and VX-702 abolished the regulatory effects of Mcl-1 on EMT. CONCLUSION: Our study underlines the importance of miR-105 in modulating NSCLC cells EMT. miR-105 promoted the EMT of NSCLC cells possibly via upregulation of Mcl-1 and thereby activation of mTOR and p38MAPK signaling.
Authors: Daniel J de Klerk; Mark J de Keijzer; Lionel M Dias; Jordi Heemskerk; Lianne R de Haan; Tony G Kleijn; Leonardo P Franchi; Michal Heger Journal: Methods Mol Biol Date: 2022
Authors: Meijie Sang; Aiying Li; Xu Wang; Can Chen; Kun Liu; Lin Bai; Ming Wu; Fei Liu; Meixiang Sang Journal: Transl Cancer Res Date: 2020-03 Impact factor: 1.241