Inhibitory effect and mechanism of hsa-miR-6832-5p on growth and metastasis of bladder tumor cells
10.3760/cma.j.cn431274-20190916-01070
- VernacularTitle:hsa-miR-6832-5p抑制膀胱肿瘤细胞生长和转移的作用及机制
- Author:
Maona ZHANG
1
;
Hong ZHANG
;
Yansha WANG
;
Sijia LYU
Author Information
1. 湖北省鄂州市中心医院病理科 436000
- From:
Journal of Chinese Physician
2020;22(9):1376-1380
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To analyze the expression of hsa-microRNA-6832-5p (hsa-miR-6832-5p) in bladder tumor tissues and bladder tumor cells, and to explore its interference on the expression of proline-rich protein 11 (PRR11) gene in bladder tumor cells and its effect on cell proliferation and migration.Methods:Real-time quantitative polymerase chain reaction (qPCR) was used to detect the expression level of hsa-miR-6832-5p in bladder tumor tissues, paratumorous tissues, bladder tumor cell lines and normal bladder epithelial cells. Bioinformatics predicts and the dual luciferase reporter gene validates the downstream target gene of hsa-miR-6832-5p. hsa-miR-6832-5p or miR-NC were transfected to the bladder tumor cell lines with the lowest expression level of hsa-miR-6832-5p, respectively, and named as miR-6832-5p group and miR-NC group. qPCR was used to detect the expression levels of hsa-miR-6832-5p and target gene mRNA in the transfected cells. Western blot was used to detect the expression level of target gene protein. Methyl thiazolyl tetrazolium (MTT) assay and transwell assay were used to detect cell proliferation and migration, respectively.Results:The expression of hsa-miR-6832-5p was lower in bladder tumor tissues than in adjacent tissues ( P<0.01). The expression of hsa-miR-6832-5p in bladder tumor cells was lower than that in normal bladder epithelial cells ( P<0.05), and T24 cells had the lowest hsa-miR-6832-5p expression level ( P<0.01). Bioinformatics and dual luciferase reporter genes showed that hsa-miR-6832-5p can directly act on the 3′-untranslated region of the PRR11 gene ( P<0.01). The expression of hsa-miR-6832-5p in the miR-6832-5p group was significantly higher than that in the miR-NC group ( P<0.01). The expression of PRR11 in the miR-6832-5p group was significantly lower than that in the miR-NC group ( P<0.01). Western blot results were consistent with qPCR results. Compared with the miR-NC group, the proliferation of bladder tumor cells was significantly decreased after transfection with hsa-miR-6832-5p ( P<0.05), and the migration ability of cells was significantly decreased ( P<0.01). Conclusions:The expression of hsa-miR-6832-5p was significantly decreased in bladder tumor tissues and cell lines. hsa-miR-6832-5p inhibited the proliferation and migration of bladder tumor cells by down-regulating the expression of PRR11 gene.
