Effects of miR-17-5p targeting B7-H3 on biological characteristics and immunomodulatory effects of colorectal cancer cells
Objective:To investigate the regulatory role of miR-17-5p on B7-H3 and its immunomodulatory role in the develop-ment of colorectal cancer.Methods:Twenty-five pairs of human colorectal cancer tissues and adjacent normal tissues were collected,and the expression levels of miR-17-5p,B7-H3 and PD-L1 were analyzed by real-time fluorescence quantitative PCR,protein immuno-blotting and immunohistochemical assays.Human colorectal cancer HCT-116 cells were divided into four groups:miR-NC group,miR-17-5p mimic group,si-NC group and si-B7-H3 group.The growth,invasive ability and apoptosis rate of HCT-116 cells were ana-lyzed by cell colony formation assay,Transwell assay and flow cytometry.The regulatory effect of miR-17-5p on B7-H3 was analyzed by luciferase reporter gene assay.The levels of IL-2,IL-4,IL-6,IL-10,IFN-γ and TNF-α cytokines in the cell culture medium were analyzed by ELISA.Results:Compared with paracancerous tissues,miR-17-5p expression was decreased and B7-H3 expression was increased in colorectal cancer tissues.miR-17-5p mimic and si-B7-H3 were able to reduce the number of HCT-116 cell colony forma-tion and invasion,promote HCT-116 cell apoptosis,and decrease the levels of IL-2,IL-4,IL-6,IL-10,IFN-γ and TNF-α cytokines in culture medium.In addition,miR-17-5p mimic and si B7-H3 were able to inhibit PD-L1 expression in HCT-116 cells.Conclusion:miR-17-5p can targeting inhibit B7-H3 expression,affect the immunomodulatory role in colorectal cancer development,and inhibit metastasis and promote apoptosis of colorectal cancer cells.
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