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miR-15b-5p靶向CDK4抑制脉络膜黑色素瘤细胞增殖

张晓楠 冯浩 白雪 应曼曼 孙胜男 宁宏

中国医科大学学报2019,Vol.48Issue(3):230-235,6.
中国医科大学学报2019,Vol.48Issue(3):230-235,6.DOI:10.12007/j.issn.0258-4646.2019.03.008

miR-15b-5p靶向CDK4抑制脉络膜黑色素瘤细胞增殖

miR-15b-5p Inhibits Choroid Melanoma Cell Proliferation by Targeting CDK4

张晓楠 1冯浩 1白雪 1应曼曼 1孙胜男 1宁宏1

作者信息

  • 1. 中国医科大学附属第一医院眼科,沈阳 110001
  • 折叠

摘要

Abstract

Objective To explore the inhibitory effects of miR-15 b-5 p on choroid melanoma cell line proliferation by targeting CDK4.Methods Dual-luciferase assay was used to verify the direct binding site between miR-15 b-5 p and CDK4 3'-UTR. MUM-2 B cells were cultured in vitro and transfected with negative control RNA, miR-15 b-5 p mimics, inhibitor normal control (nc) RNA, and miR-15 b-5 p inhibitor. qRT-PCR was used to detect miR-15 b-5 p expression, Western blotting was used to measure the expression levels of CDK4 in the cells, CCK-8 assay was used to detect proliferation capacity, and flow cytometry was used to detect cell cycle. Results Dual-luciferase assay verified that miR-15 b-5 p could bind to CDK4 mRNA 3'-UTR successfully. Compared to the negative control group, the mimics group showed significantly increased miR-15 b-5 p expression, decreased CDK4 levels, decreased cell proliferation rate, and increased proportion of G1-phase cells. Compared to the inhibitor nc group, the inhibitor group showed significantly decreased miR-15 b-5 p expression (t = 25.01, P < 0.000 1), increased CDK4 protein level, increased cell proliferation rate, and decreased proportion of G1-phase cells.Conclusion miR-15 b-5 p can target CDK4, induce G1 phase arrest in cells, and thus, reduce the proliferation rate of choroid melanoma cells.

关键词

脉络膜黑色素瘤/细胞周期素依赖性激酶4/miR-15b-5p

Key words

choroidal melanoma/CDK4/miR-15b-5p

分类

医药卫生

引用本文复制引用

张晓楠,冯浩,白雪,应曼曼,孙胜男,宁宏..miR-15b-5p靶向CDK4抑制脉络膜黑色素瘤细胞增殖[J].中国医科大学学报,2019,48(3):230-235,6.

基金项目

辽宁省自然科学基金(20180551171) (20180551171)

中国医科大学学报

OA北大核心CSCDCSTPCD

0258-4646

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