南方医科大学学报

南方医科大学学报 ›› 2022, Vol. 42 ›› Issue (5): 712-717.doi: 10.12122/j.issn.1673-4254.2022.05.12

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MiR-744-5p通过靶向 CCND1 抑制肾透明细胞癌细胞的增殖、侵袭和迁移

雷坤阳，谢文杰，孙 庭，刘贻富，王 旭   

  1. 南昌大学第一附属医院泌尿外科，病理科，江西 南昌 330006
  • 出版日期:2022-05-20 发布日期:2022-06-02

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

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摘要： 目的 探讨miR-744-5p/CCND1轴在肾透明细胞癌(ccRCC)中的作用及机制。方法 qRT-PCR检测ccRCC组织和细胞系中miR-744-5p 的表达水平。实验分组：miR-744-5p 模拟物（miR-744-5p mimic）、阴性对照（NC mimic）、CCND1 模拟物（CCND1 mimic）及其阴性对照（NC mimic）。CCK-8实验、划痕愈合实验和transwell实验验证miR-744-5p对ccRCC细胞增殖、迁移及侵袭功能的影响。通过生物信息学预测miR-744-5p的下游靶分子，qRT-PCR和Western blot验证CCND1在786-O及OSRC2细胞中的表达水平，双荧光素酶报告基因实验进一步验证miR-744-5p和CCND1的关系，最后通过回复实验验证miR-744-5p/CCND1轴在ccRCC中的作用。结果 MiR-744-5p在ccRCC组织和细胞系中显著下调（P<0.05），过表达miR-744-5p可以抑制ccRCC细胞的增殖、迁移和侵袭（P<0.05）。生物信息学分析结果和双荧光素酶报告基因实验结果显示，CCND1是miR-744-5p的下游靶标。回复实验结果显示，上调CCND1可以部分逆转上调miR-744-5p对ccRCC细胞增殖、迁移以及侵袭的抑制作用（P<0.05）。结论 miR-744-5p通过靶向CCND1抑制ccRCC细胞的恶性表型，miR-744-5p/CCND1轴可能是ccRCC诊断和治疗的一个新的靶点。

关键词: 肾透明细胞癌；miR-744-5p；CCND1

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