| 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 |
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| 引用本文： | 雷坤阳,谢文杰,孙庭,刘贻富,王旭.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[J].南方医科大学学报,2022,42(5):712-717. |
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| 作者姓名： | 雷坤阳 谢文杰 孙庭 刘贻富 王旭 |
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| 作者单位： | 南昌大学第一附属医院泌尿外科，病理科，江西 南昌 330006 |
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| 基金项目： | 江西省卫生健康委科技计划 |
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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。
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| 关 键 词： | 肾透明细胞癌 miR-744-5p CCND1 |
MiR-744-5p inhibits the proliferation,invasion, and migration of clear-cell renal cell carcinoma cells by targeting CCND1 |
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| LEI Kunyang,XIE Wenjie,SUN Ting,LIU Yifu,WANG Xu.MiR-744-5p inhibits the proliferation,invasion, and migration of clear-cell renal cell carcinoma cells by targeting CCND1[J].Journal of Southern Medical University,2022,42(5):712-717. |
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| Authors: | LEI Kunyang XIE Wenjie SUN Ting LIU Yifu WANG Xu |
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| Affiliation: | Department of Urology, Department of Pathology, First Affiliated Hospital of Nanchang University, Nanchang 330006, China |
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| 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. |
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| Keywords: | clear-cell renal cell carcinoma miR-744-5p CCND1 |
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