|Table of Contents|

Study on the mechanism of miR-335-5p regulating the immune escape of prostate cancer cells through targeted regulation on Galectin-3

Journal Of Modern Oncology[ISSN:1672-4992/CN:61-1415/R]

Issue:
2023 13
Page:
2431-3437
Research Field:
Publishing date:

Info

Title:
Study on the mechanism of miR-335-5p regulating the immune escape of prostate cancer cells through targeted regulation on Galectin-3
Author(s):
ZHOU Rubao; GUO Zonghua; YANG Jun; XU Wei; ZHAO Kedong; RAO Zhigang; KONG Dongbo; JIANG Botao
Department of Urology,Xianning Central Hospital(The First Affiliated Hospital of Hubei University of Science and Technology),Hubei Xianning 437199,China.
Keywords:
miR-335-5p; prostate cancer; galectin-3; immune escape
PACS:
R735.25
DOI:
10.3969/j.issn.1672-4992.2023.13.011
Abstract:
Objective:To investigate the effect of miR-335-5p on the immune escape of prostate cancer cells through targeted regulation on Galectin-3.Methods:qRT-PCR was used to detect the expression of miR-335-5p in normal prostate epithelial cells WPMY-1 and prostate cancer cells LNCaP,PC-3,DU145.PC-3 cells were divided into:NC group,mimics NC group,miR-335-5p mimics group,miR-335-5p mimics+pcDNA group,and miR-335-5p mimics+pcDNA-Galectin-3 group,qRT-PCR was performed to measure the expression of miR-335-5p of cells in each group.Western blot was performed to measure the expression of Galectin-3 protein of cells in each group.After the cells in above five groups were co-cultured with the natural killer cell NK-92MI,ELISA method was performed to measure the levels of tumor necrosis factor alpha(TNF-α) and interferon gamma(IFN-γ) in the supernatant of co-cultured cells.CCK-8 method was performed to measure the immune killing rate of NK-92MI cells.The apoptosis rate of NK-92MI cells was detected by flow cytometry.The targeting relationship between miR-335-5p and Galectin-3 was verified by dual-luciferase reporter gene,RNA pull-down,and RNA-binding protein immunoprecipitation(RIP) experiments.Results:Compared with normal prostate epithelial cells WPMY-1,the expression level of miR-335-5p in prostate cancer cells LNCaP,PC-3,and DU145 was significantly reduced(P＜0.05),and the expression level of miR-335-5p in PC-3 cells was the lowest,so PC-3 cells were selected as the follow-up research object.Compared with the NC group,the expression level of miR-335-5p in PC-3 cells in the miR-335-5p mimics group was significantly increased,and the expression of Galectin-3 protein was significantly decreased(P＜0.05).Compared with the NC co-culture group,the levels of TNF-α and IFN-γ in the cell supernatant and the immune killing rate of NK-92MI cells in the miR-335-5p mimics co-culture group were significantly increased,reduced apoptosis rate of NK-92MI cells(P＜0.05).Compared with the miR-335-5p mimics group,there was no significant difference in the expression level of miR-335-5p in PC-3 cells in the miR-335-5p mimics+pcDNA-Galectin-3 group(P>0.05),the expression of Galectin-3 protein was significantly increased(P＜0.05).Compared with the miR-335-5p mimics co-culture group,the levels of TNF-α and IFN-γ in the cell supernatant and the immune killing rate of NK-92MI cells in the miR-335-5p mimics+pcDNA-Galectin-3 co-culture group were significantly decreased,increased apoptosis rate of NK-92MI cells(P＜0.05).miR-335-5p targeted and negatively regulated the Galectin-3 expression.Conclusion:Overexpression of miR-335-5p can inhibit the immune escape of PC-3 cells by targeting down-regulation of Galectin-3 expression.

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Memo

Memo:
湖北省卫生健康委2021-2022年度面上项目(编号：WJ2021M091)
Last Update: 2023-05-31