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  

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

作　　者：雷坤阳 谢文杰[1] 孙庭[1] 刘贻富 王旭 LEI Kunyang;XIE Wenjie;SUN Ting;LIU Yifu;WANG Xu(Department of Urology,First Affiliated Hospital of Nanchang University,Nanchang 330006,China;Department of Pathology,First Affiliated Hospital of Nanchang University,Nanchang 330006,China)

机构地区：[1]南昌大学第一附属医院泌尿外科,江西南昌330006 [2]南昌大学第一附属医院病理科,江西南昌330006

出　　处：《南方医科大学学报》2022年第5期712-717,共6页Journal of Southern Medical University

基　　金：江西省卫生健康委科技计划(202210397)。

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Westernblot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。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.

关 键 词：肾透明细胞癌 miR-744-5p CCND1 

分 类 号：R737.11[医药卫生—肿瘤]