miR-502调控犬乳腺肿瘤细胞增殖、迁移和EMT的作用机制

doi:10.11843/j.issn.0366-6964.2023.10.034

Abstract

Abstract: This study aimed to investigate the effect and mechanism of microRNA-502(miR-502) targeting RBMS1 on proliferation, migration and epithelial-mesenchymal transition (EMT) of canine mammary tumor cells. The transcription level of miR-502 and RBMS1 mRNA in canine mammary tumor cell lines (CHMp and CHMm) and normal mammary gland tissues were detected by realtime quantitative PCR (qRT-PCR). The miR-502 mimic/inhibitor and negative control mimic/inhibitor NC were transfected into CHMp/CHMm cells using liposome transfection method,respectively,and the expression levels of miR-502 in cells were detected by qRT-PCR method. MTT assay and Transwell assay were used to detect the proliferation and migration ability of CHMm cells; the expression levels of epithelial cell marker E-cadherin, mesenchymal cell marker vimentin, cell migration markers matrix metalloproteinase (MMP2) and cell proliferation marker PCNA after transfection with miR-502 were detected by qRT-PCR and Western blot methods. qRT-PCR and double-luciferase reporter gene assay were adopted to verify the targeting relationship between miR-502 and RBMS1 gene. The results showed that compared with normal mammary tissues, the expression level of miR-502 in canine mammary tumor cells was significantly up-regulated, while RBMS1 was down-regulated. Transfection of miR-502 mimic up-regulated the expression level of miR-502, whereas miR-502 inhibitor down-regulated the expression level of miR-502. Compared with the miR-NC group, transfection of miR-502 mimic significantly promoted the proliferation and migration of CHMm cells, moreover, down-regulated the expression of E-cadherin mRNA and protein, and up-regulated the expression of vimentin, MMP2, PCNA mRNA and protein. On the contrary, transfection of miR-502 inhibitor significantly suppressed the proliferation and migration of CHMm cells, up-regulated the expression of E-cadherin and down-regulated the expression of vimentin, MMP2, PCNA mRNA and protein; qRT-PCR and luciferase report experiments showed that miR-502 inhibited the expression of RBMS1 by targeting RBMS1 3'-UTR. Therefore, miR-502 is highly expressed in canine mammary tumor cells, and interfering with its expression might affected the proliferation, migration and EMT of the canine mammary tumor CHMm cells, and miR-502 targeted regulation the expression of RBMS1 mRNA.

Key words: miR-502, RBMS1, canine mammary tumor cells, cell proliferation/migration, epithelial-mesenchymal transition

CLC Number:

S858.2927.26

REN Xiaoli, FAN Yuying, HUANGFU Heping, WANG Jun, JIN Shuangxing, LIU Yun, SHI Dongmei. Mechanism of miR-502 Regulating Proliferation, Migration, Invasion and EMT of Canine Mammary Tumor Cells[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(10): 4379-4388.

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Mechanism of miR-502 Regulating Proliferation, Migration, Invasion and EMT of Canine Mammary Tumor Cells

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[1]	REN Xiaoli, FAN Yuying, HUANGFU Heping, LIU Yun, SHI Dongmei. Effect of GSK126 on Epithelial-mesenchymal Transition of Canine Mammary Tumor Cells [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(4): 1721-1729.
[2]	REN Xiaoli, FAN Yuying, SHI Dongmei, TIAN Chuanwen, LU Shuilong, LIU Yun. Expressions of miR-502 in Canine Breast Cancer and Clinical Significance [J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(1): 193-197.