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Archive > Volume 28 Issue 8 > 2021,28(8):824-832. DOI:10.3872/j.issn.1007-385X.2021.08.009 Prev Next
Clinical Research
Highly expressed miR-504 in gastric cancer tissues regulates the biological behaviors of gastric cancer cell BGC-823 through TP53INP1
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Abstract:
Objective: To explore the expression level of microRNA-504 (miRNA-504) in gastric cancer (GC) tissues and its regulatory mechanism in the biological behaviors of GC cells. Methods: From June 2020 to December 2020, tumor tissues and corresponding para-cancerous tissues resected from 48 patients with gastric cancer were collected in the Department of Surgical, Sanya Central Hospital. qPCR was used to detect the mRNA levels of miR-504 and tumor protein 53-induced nuclear protein 1 (TP53INP1) in the tissues, and WB assay was used to detect the protein level of TP53INP1. Human gastric cancer BGC-823 cells were cultured in vitro and divided into control group (normally cultured BGC-823 cells), miR-504 mimic group, mimic-NC group, miR-504 inhibitor group,inhibitor-NC group, miR-504 inhibitor+si-NC group, and miR-504 inhibitor+si-TP53INP1 group. qRT-PCR was used to detect mRNA expression of miR-504 and TP53INP1 in each group of cells. MTT method, Flow cytometry, Scratch test, and Transwell invasion assay were used to detect the proliferation, apoptosis, migration, and invasion of the cells in each group. WB assay was used to detect the protein expression of cell proliferation, migration, and invasion-related proteins (Cyclin D1, E-cadherin, MMP-2, MMP-9) and TP53INP1 in each group. Dual-luciferase reporter gene experiment was used to further verify the targeting relationship between miR[1]504 and TP53INP1. Results: Compared with the para-cancerous tissues, the expression of miR-504 in gastric cancer tissues was significantly increased (P<0.05), and the mRNA and protein expression levels of TP53INP1 were significantly decreased (P<0.05 or P <0.01). The expression of miR-504 was negatively correlated with TP53INP1 mRNA (P<0.01). Compared with the control group, the expression of miR-504 in the BGC-823 cells of the miR-504 mimic group was significantly increased, while the mRNA and protein expression of TP53INP1 was significantly decreased (all P<0.05); moreover, the cell survival rate, scratch healing rate, the number of cells entering the lower layer of the Transwell chamber, and the expression of Cyclin D1, MMP-2, and MMP-9 were all significantly increased, while the apoptosis rate and the expression of E-cadherin were significantly decreased (all P<0.05) in miR-504 mimic group;However, the miR-504 inhibitor could significantly down-regulate the expression of miR-504, up-regulate the mRNA and protein expression of TP53INP1, inhibit cell proliferation, migration, and invasion, and promote cell apoptosis (all P<0.05) in BGC-823 cells.Down-regulating the expression of TP53INP1 could significantly reduce the inhibitory effect of down-regulation of miR-504 on the proliferation, migration, and invasion of BGC-823 cells; and miR-504 over-expression could obviously inhibit the luciferase activity of wild-type TP53INP1 plasmid (all P<0.05). Conclusion: miR-504 is highly expressed in gastric cancer tissues. Down-regulation of miR[1]504 can inhibit the malignant biological behaviors of gastric cancer BGC-823 cells and promote their apoptosis, and the mechanism may be related to the targeted regulation of the expression of TP53INP1.
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Project Supported:
Project supported by Scientific Research Project of Health and Family Planning Industry in Hainan Province (No.16A200142), and the Sanya Medical and Health Science and Technology Innovation Project (No. 2017YW08)
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