Journal of Southern Medical University ›› 2022, Vol. 42 ›› Issue (5): 712-717.doi: 10.12122/j.issn.1673-4254.2022.05.12

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MiR-744-5p inhibits the proliferation, invasion, and migration of clear-cell renal cell carcinoma cells by targeting CCND1

LEI Kunyang, XIE Wenjie, SUN Ting, LIU Yifu, WANG Xu   

  1. Department of Urology, Department of Pathology, First Affiliated Hospital of Nanchang University, Nanchang 330006, China
  • Online:2022-05-20 Published:2022-06-02

Abstract: Objective To explore the role of miR-744-5p/CCND1 axis in clear-cell renal cell carcinoma (ccRCC). Methods We examined the expression levels of miR-744-5p in 65 pairs of ccRCC and adjacent tissue specimens and in 5 ccRCC cell lines and human renal tubular epithelial (HK2) cells using qRT-PCR. The ccRCC cell lines 786-O and OSRC2 were transfected with miR-744-5p mimic, CCND1 mimic, or their negative control mimics, and the changes in cell proliferation, migration, and invasion were evaluated with CCK-8, wound healing, and Transwell assays. The downstream target molecules of miR-744-5p were predicted by bioinformatics analysis, and the expression level of CCND1 in ccRCC cells was verified by qRT-PCR and Western blotting. The relationship between miR-744-5p and CCND1 was further validated by dual luciferase reporter assay, and the role of the miR-744-5p/CCND1 axis in ccRCC was explored by rescue experiments. Results MiR-744-5p was significantly downregulated in ccRCC tissues and cell lines (all P<0.05), and its overexpression inhibited the proliferation, migration, and invasion of ccRCC cells (all P<0.05). Bioinformatics analysis and dual luciferase reporter assay showed that CCND1 was a downstream target of miR-744-5p. The results of rescue experiments showed that upregulation of CCND1 could partially reverse the inhibitory effect of miR-744-5p overexpression on ccRCC cell proliferation, migration, and invasion (all P<0.05). Conclusion MiR-744-5p inhibits the malignant phenotype of ccRCC cells by targeting CCND1, and the miR-744-5p/CCND1 axis may be a novel target for diagnosis and treatment of ccRCC.

Key words: clear-cell renal cell carcinoma; miR-744-5p; CCND1