Int J Med Sci 2021; 18(8):1798-1809. doi:10.7150/ijms.50871 This issue

Research Paper

MicroRNA-362 negatively and positively regulates SMAD4 expression in TGF-β/SMAD signaling to suppress cell migration and invasion

Han Ping Cheng1,2,7, Chiu-Jung Huang6, Ming-Long Tsai5, Hooi Tin Ong3, Soon Keng Cheong1,4, Kong Bung Choo1,3✉, Shih-Hwa Chiou5,7,8✉

1. Centre for Stem Cell Research, Faculty of Medicine and Health Sciences, Universiti Tunku Abdul Rahman, Selangor, Malaysia.
2. Postgraduate Program, Faculty of Medicine and Health Sciences, Universiti Tunku Abdul Rahman, Selangor, Malaysia.
3. Department of Preclinical Sciences, Faculty of Medicine and Health Sciences, Universiti Tunku Abdul Rahman, Selangor, Malaysia.
4. Dean's Office, Faculty of Medicine and Health Sciences, Universiti Tunku Abdul Rahman, Selangor, Malaysia.
5. Department of Medical Research, Taipei Veterans General Hospital, Taipei, Taiwan.
6. Department of Animal Science & Graduate Institute of Biotechnology, Chinese Culture University, Taipei, Taiwan.
7. Institutes of Pharmacology, National Yang-Ming University, Taipei, Taiwan.
8. Institute of Clinical Medicine, National Yang-Ming University, Taipei, Taiwan.

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Citation:
Cheng HP, Huang CJ, Tsai ML, Ong HT, Cheong SK, Choo KB, Chiou SH. MicroRNA-362 negatively and positively regulates SMAD4 expression in TGF-β/SMAD signaling to suppress cell migration and invasion. Int J Med Sci 2021; 18(8):1798-1809. doi:10.7150/ijms.50871. Available from https://www.medsci.org/v18p1798.htm

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Abstract

Graphic abstract

Cell migration and invasion are modulated by epithelial-to-mesenchymal transition (EMT) and the reverse MET process. Despite the detection of microRNA-362 (miR-362, both the miR-362-5p and -3p species) in cancers, none of the identified miR-362 targets is a mesenchymal or epithelial factor to link miR-362 with EMT/MET and metastasis. Focusing on the TGF-β/SMAD signaling pathway in this work, luciferase assays and western blot data showed that miR-362 targeted and negatively regulated expression of SMAD4 and E-cadherin, but not SNAI1, which is regulated by SMAD4. However, miR-362 knockdown also down-regulated SMAD4 and SNAI1, but up-regulated E-cadherin expression. Wound-healing and transwell assays further showed that miR-362 knockdown suppressed cell migration and invasion, effects which were reversed by over-expressing SMAD4 or SNAI1, or by knocking down E-cadherin in the miR-362 knockdown cells. In orthotopic mice, miR-362 knockdown inhibited metastasis, and displayed the same SMAD4 and E-cadherin expression profiles in the tumors as in the in vitro studies. A scheme is proposed to integrate miR-362 negative regulation via SMAD4, and to explain miR-362 positive regulation of SMAD4 via miR-362 targeting of known SMAD4 suppressors, BRK and DACH1, which would have resulted in SMAD4 depletion and annulment of subsequent involvement in TGF-β signaling actions. Hence, miR-362 both negatively and positively regulates SMAD4 expression in TGF-β/SMAD signaling pathway to suppress cell motility and invasiveness and metastasis, and may explain the reported clinical association of anti-miR-362 with suppressed metastasis in various cancers. MiR-362 knockdown in miR-362-positive cancer cells may be used as a therapeutic strategy to suppress metastasis.

Keywords: microRNA-362, EMT/MET, SMAD4, TGF-β/SMAD signaling pathway, cell migration/invasion, metastasis.