Copyright (c) 2023 Xiaowei Qin, Jinjie Zhang, Mengmeng Wang, Yafei Feng, Yanbo Zhao, Huipeng Zhang, Wei Guo
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The undersigned hereby assign all rights, included but not limited to copyright, for this manuscript to CMB Association upon its submission for consideration to publication on Cellular and Molecular Biology. The rights assigned include, but are not limited to, the sole and exclusive rights to license, sell, subsequently assign, derive, distribute, display and reproduce this manuscript, in whole or in part, in any format, electronic or otherwise, including those in existence at the time this agreement was signed. The authors hereby warrant that they have not granted or assigned, and shall not grant or assign, the aforementioned rights to any other person, firm, organization, or other entity. All rights are automatically restored to authors if this manuscript is not accepted for publication.The effect of thalidomide on the invasive ability of gastric cancer cells by regulating miR-524-5p/FSTL1
Corresponding Author(s) : Wei Guo
Cellular and Molecular Biology,
Vol. 69 No. 14: Cancer molecular biology: Diagnosis and treatment
Abstract
This study aimed to investigate the effect of thalidomide (Thal) regulating microRNA (miR)-524-5p/follistatin-like protein 1 (FSTL1) on the invasion ability of gastric cancer cells. For this purpose, real-time fluorescent quantitative PCR (RT-qPCR) was used to detect the level of miR-524-5p in GES1, SUN-16, MGC-803, SGC-7901, MKN-28, and MKN-45 cells. Then the MGC-803 and MKN-45 cells would proceed to the next research. The MGC-803 and MKN-45 cells were cultured in vitro and added Thal to the final concentration of (6.25, 12.5, 25, 50, 100) μg/mL. The blank control group only added 0.1% dimethyl sulfoxide (DMSO) culture medium, and cultured for 48 hours. CCK8 was used to detect cell proliferation, and Transwell was used to detect cell invasion. The experiment was divided into a blank control group, Thal group (25 μg/mL Thal-treated cells), Thal+inhibitor NC group, and Thal+miR-524-5p inhibitor group (transfected with inhibitor NC and miR-524-5p inhibitor respectively on the basis of Thal group), cultivated for 48 h. The level of miR-524-5p in the cells was detected by RT-qPCR; the cell invasion was detected by Transwell; the expression of matrix metalloproteinase (MMP)-2, MMP-9, FSTL1 protein in the cells was detected by Western blot. The targeting relationship between miR-524-5p and FSTL1 was verified by dual luciferase. Results showed that compared with GES1 cells, the level of miR-524-5p in SUN-16, MGC-803, SGC-7901, MKN-28, and MKN-45 cells decreased (P<0.05). In MGC-803 and MKN-45 cells, compared with the blank control group, the cell proliferation rate and the number of invasions in the (50, 100) μg/mL Thal treatment groups, and the number of invasions in (6.25, 12.5, 25) μg/mL Thal treatment groups decreased (P<0.05). Compared with the blank control group, the level of miR-524-5p in the cells of the Thal group, Thal+inhibitor NC group, and Thal+miR-524-5p inhibitor group increased (P<0.05), the number of invasions, the levels of MMP-2, MMP-9 and FSTL1 proteins in cells decreased (P<0.05); compared with the Thal group and the Thal+inhibitor NC group, the level of miR-524-5p in the cells of the Thal+miR-524-5p inhibitor group decreased (P<0.05), the number of invasions, the levels of MMP-2, MMP-9, and FSTL1 proteins in the cells increased (P<0.05). Dual luciferase verification revealed that there was a targeting relationship between miR-524-5p and FSTL1. In conclusion, that can up-regulate the expression of miR-524-5p to reduce the expression of FSTL1 protein, inhibit the invasion of gastric cancer cells, and achieve alleviation of the disease.
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