Mobile website
Archive > Volume 29 Issue 8 > 2022,29(8):732-740. DOI:10.3872/j.issn.1007-385X.2022.08.005 Prev Next
Basic Research
miR-124 affects malignant biological behaviours of renal cell carcinoma OS-RC-2 cells by regulating the Jagged1/Notch signaling pathway
- Article
- Figures
- Metrics
- Preview PDF
- Reference
- Related
- Cited by
- Materials
Abstract:
Objective: To investigate the effects of miR-124 on the proliferation, apoptosis, migration and invasion of renal cell carcinoma (RCC) cells by regulating the Jagged1 (JAG1)/Notch signaling pathway. Methods: The RCC tissues and paracancerous tissues of 38 RCC patients treated in Wuhan Third Hospital from June 2018 to October 2021 were collected, and RCC cells (Caki-2,A498, ACHN, 786-O, OS-RC-2) and human normal kidney cells (293T) were cultured in vitro. The expression levels of miR-124 and JAG1 proteins in RCC tissues and cells were detected by immunohistochemistry, qPCR and WB assay. OS-RC-2 cells, which had the greatest difference in expression of miR-124 with that of 293T cells, were selected for transfection and divided into control group, NC mimic group, miR-124 mimic group, miR-124 mimic+pcDNA group and miR-124 mimic+pc-JAG1 group according to the difference in transfectants. The relationship between miR-124 and JAG1 was verified by dual-luciferase reporter gene experiments; the expressions of miR-124 and JAG1 mRNA were detected by qPCR; the protein expression of JAG1 was analyzed by immunohistochemistry method; WB assay was performed to measure the expressions of JAG1, apoptosis-related proteins (cleaved caspase-3, BAX, Bcl2) and Notch signaling pathway-related proteins (NICD, HES1 and HES5); MTT method was performed to measure the proliferation of OS-RC-2 cells; Transwell assay was performed to measure the migration and invasion of OS-RC-2 cells;and flow cytometry was performed to measure the apoptosis of OS-RC-2 cells. Results:Compared with paracancerous tissues, the expression of miR-124 in RCC tissues decreased, and the expressions of JAG1 mRNA and protein increased (all P<0.01); compared with 293T cells, the levels of miR-124 decreased while the expressions of JAG1 mRNA and protein increased in Caki-2, A498, ACHN,786-O and OS-RC-2 cells (all P<0.05); miR-124 directly negatively regulated JAG1. Compared with the Control group and the NC mimic group, the expression level of miR-124, the apoptosis rate and the expressions of cleaved caspase-3 and BAX proteins in the miR-124 mimic group increased; the expressions of JAG1 mRNA and protein decreased; the cell viability (24, 48, 72 h) decreased; the numbers of migrating and invasive cells decreased; the expressions of Bcl2 and NICD, HES1, HES5 proteins decreased (all P<0.05).Compared with the miR-124 mimic+pcDNA group and the miR-124 mimic group, the expression level of miR-124, the apoptosis rate and the expressions of cleaved caspase-3 and BAX proteins in the miR-124 mimic+pc-JAG1 group decreased; the expressions of JAG1 mRNA and protein increased; the cell viability (24, 48, 72 h) increased; the numbers of migrating and invasive cells increased and the expressions of Bcl2 and NICD, HES1, HES5 proteins increased (all P<0.05). Conclusion: miR-124 reduced the proliferation, migrationand invasion abilities of RCC OS-RC-2 cells and promoted apoptosis by downregulating JAG1 and inhibiting the Notch signaling pathway.
Keywords:
Project Supported:
Clc Number:
