Mobile website
Archive > Volume 27 Issue 5 > 2020,27(5):477-486. DOI:10.3872/j.issn.1007-385X.2020.05.002 Prev Next
Basic Research
miR-625 inhibits the malignant biological behavior of non-small cell lung cancer cells by negatively regulating Resistin expression and its mechanisms
- Article
- Figures
- Metrics
- Preview PDF
- Reference
- Related
- Cited by
- Materials
Abstract:
Objective: To investigate the effects of miR-625 and Resistin on the proliferation, invasion and migration of NSCLC cells as well as the growth of transplanted tumors in nude mice and their possible mechanisms. Methods: qPCR was used to detect the expression of miR-625 and Resistin in 80 pairs of NSCLC and corresponding para-cancerous tissues (specimens collected from NSCLC patients who were surgically treated in Xinjiang Uygur Autonomous Region People’s Hospital from March 2018 to October 2019) and four cell lines. Bioinformatics was adopted to predict the targeting relationship between miR-625 and Resistin, which was then verified by Dual luciferase gene reporter experiment. Overexpression or inhibition of miR-625 and Resistin in NSCLC cells was achieved with lipofection transfection technology, and the experimental cells were divided into miR-625 mimic group, miR-625 inhibitor group, si-Resisitin group, miR-625 inhibitor+si-Resisitin group and NC group. The effects of miR-625 and Resistin on proliferation, invasion and migration of NSCLC cells were detected by CCK-8, Transwell and Scratch test, respectively. Western blotting was used to detect the effects of miR-625 and Resistin on the expressions of PI3K/AKT/Snail pathway proteins related with EMT in NSCLC cells. A549 cell transplanted tumor model was constructed in nude mice to observe the effect of miR-625 and Resistin on the growth of xenografts. Results: Compared with para-cancerous tissues, miR-625 showed low expression while Resistin showed high expression in NSCLC tissues and four cell lines (both P<0.01), and the two were negatively correlated (r=-0.7183,P<0.01). The expression of Resistin was related to the degree of NSCLC differentiation, clinical stage and lymph node metastasis. Resistin was a target gene of miR-625. Compared with the Blank group and NC group, the proliferation, invasion and migration of NSCLC cell lines A549 and H226, as well as the growth of transplanted tumors in nude mice in the miR-625 mimic group and the si-Resistin group were significantly reduced (all P<0.05), while those indicators in the miR-625 inhibitor group were significantly improved (all P<0.05); However, co-transfection of miR-625 inhibitor and si-Resistin significantly reversed the effect of miR-625 inhibitor on above indicators (all P<0.05); And there was no significant difference between NC group and miR-625 inhibitor+si-Resistin group (all P>0.05). The protein expressions of p-AKT, p-PI3K, Snail, Twist1 and Vimentin also showed the same trend (all P<0.05), while the expression of E-cadherin protein changed in the opposite direction (P<0.05). Conclusion: miR-625 is lowly expressed in NSCLC tissues and cell lines, which can negatively regulate Resistin expression to inhibit the proliferation, invasion and migration of NSCLC cells and the growth of transplanted tumors in nude mice. The mechanism may be related to the PI3K/AKT/Snail signaling pathway.
Keywords:
Project Supported:
Project supported by the Natural Science Foundation of Xinjiang Uygur Autonomous Region (No. 2019D01C108)
Clc Number:
