OncoTargets and Therapy (Aug 2021)
Long Non-Coding RNA TRG-AS1 Promoted Proliferation and Invasion of Lung Cancer Cells Through the miR-224-5p/SMAD4 Axis
Abstract
Mengyan Zhang,1,2,* Weiguo Zhu,3,* Mansour Haeryfar,4 Sumei Jiang,5 Xiang Jiang,6 Wei Chen,7 Jiancheng Li2 1The School of Clinical Medicine, Fujian Medical University, Fuzhou, People’s Republic of China; 2Department of Radiation Oncology, Fujian Medical University Cancer Hospital & Fujian Cancer Hospital, Fuzhou, 350014, People’s Republic of China; 3Department of Radiotherapy, The Affiliated Huaian No.1 People’ s Hospital of Nanjing Medical University, Huaian, Jiangsu Province, 223300, People’s Republic of China; 4Department of Microbiology and Immunology, Western University, London, Ontario, N6A 3K7, Canada; 5Department of B-ultrasound, The Affiliated Huaian No.1 People’ s Hospital of Nanjing Medical University, Huaian, Jiangsu Province, 223300, People’s Republic of China; 6Department of Hernia Surgery, The Affiliated Huaian No.1 People’ s Hospital of Nanjing Medical University, Huaian, Jiangsu Province, 223300, People’s Republic of China; 7Department of Respiratory Medicine, The Affiliated Huaian No.1 People’ s Hospital of Nanjing Medical University, Huaian, Jiangsu Province, 223300, People’s Republic of China*These authors contributed equally to this workCorrespondence: Jiancheng LiDepartment of Radiation Oncology, Fujian Medical University Cancer Hospital & Fujian Cancer Hospital, No. 420, Fuma Road, Jin’an District, Fuzhou City, Fujian Province, 350014, People’s Republic of ChinaEmail [email protected]: The aim of this study was to investigate the role and mechanism of long non-coding RNA (lncRNA) TRG-AS1 in mediating the proliferation, invasion and migration of lung cancer cells as well lung tumor growth.Methods: Firstly, the expression levels of TRG-AS1, miR-224-5p in lung cancer tissues or cells were quantified by quantitative real-time PCR. Western blot analysis was conducted to measure the expression levels of protein SMAD4. CCK-8 assay, wound healing assay and transwell assay were conducted to evaluate cell proliferation, migration and invasion, respectively. The interaction between TRG-AS1 and miR-224-5p was predicted by bioinformatics analysis. Dual-luciferase assay and RNA pull-down assay were performed to further confirm their interaction. In addition, the interaction between miR-224-5p and SMAD4 was detected by RIP assay.Results: The results showed that TRG-AS1 was highly upregulated and miR-224-5p was downregulated in lung cancer. A negative correlation was found between TRG-AS1 and miR-224-5p. Furthermore, upregulation of TRG-AS1 promoted cell proliferation and invasion, while overexpression of miR-224-5p attenuated the effects of TRG-AS1. The downstream protein SMAD4 played an important role. In vivo study showed that knockdown of TRG-AS1 effectively retarded tumor growth.Discussion: Our data suggested that the TRG-AS1/miR-224-5p/SMAD4 axis may be a potential therapeutic target in lung cancer.Keywords: lung cancer, TRG-AS1, miR-224-5p/SMAD4 axis, therapeutic target
