MiR-5586-5p通过多重靶向乳腺癌中的HIF-1α、HBEGF和ADAM17抑制缺氧诱导的血管生成。

MiR-5586-5p Suppresses Hypoxia-induced Angiogenesis Through Multiple Targeting of HIF-1α, HBEGF and ADAM17 in Breast Cancer.

作者信息

Shin Dongjo, Yoo Je-Ok, Jeong Jae-Hoon, Han Young-Hoon

机构信息

Division of Radiation Biomedical Research, Korea Institute of Radiological and Medical Sciences (KIRAMS), Seoul, Republic of Korea.

Radiological and Medico-Oncological Sciences, University of Science and Technology (UST), Daejeon, Republic of Korea.

出版信息

Anticancer Res. 2025 Feb;45(2):473-489. doi: 10.21873/anticanres.17437.

DOI:10.21873/anticanres.17437

PMID:39890164

Abstract

BACKGROUND/AIM: Hypoxia-inducible factor-1 alpha (HIF-1α) plays a key role in the cellular response to hypoxia, which plays a crucial role in the induction of abnormal angiogenesis and metastasis. Understanding the mechanism for the regulation of angiogenesis by HIF-1α-regulating miRNA will contribute to developing the strategy to prevent metastasis.

MATERIALS AND METHODS

We conducted a functional screening for HIF-1α-inhibiting miRNAs by evaluating the effects of miRNA mimics on HIF-1α expression and identified miR-5586-5p as an angiogenesis inhibitor through a mechanistic study. Angiogenic activity was assessed by tube formation assays using HUVEC cells exposed to conditioned media from miRNA-transfected breast cancer cells. In vivo activity of miR-5586-5p was examined through intratumoral injection of miRNA in orthotopic xenograft mice established by injecting MDA-MB-231 cells into the mammary fat pads of BALB/c nu/nu mice.

RESULTS

The expression of the critical proangiogenic factors vascular endothelial growth factor A (VEGFA) and angiopoietin-like protein 4 (ANGPTL4) was inhibited by miR-5586-5p. Migration and tube formation of human umbilical vein endothelial cells were reduced in the conditioned medium prepared from miR-5586-5p-transfected cells. miR-5586-5p also suppressed the expression of heparin-binding EGF-like growth factor (HBEGF) and a disintegrin and metalloprotease 17 (ADAM17), which play a role in hypoxic signaling to induce the expression of VEGFA and ANGPTL4. HIF-1α, HBEGF, and ADAM17 were verified as the direct targets of miR-5586-5p responsible for the angiogenesis-suppressing function of miR-5586-5p. Expression levels of miR-5586-5p were lower in tumor tissues than in neighboring normal tissues of breast cancer patients. The expression of miR-5586-5p was inversely correlated to those of HIF-1α, HBEGF, ADAM17, VEGFA, and ANGPTL4. Angiogenesis and subsequent tumor growth were suppressed by intratumoral injection of miR-5586-5p in orthotopic MDA-MB-231 xenografts in mice.

CONCLUSION

A potent tumor-suppressive function of miR-5586-5p applicable for the development of a novel cancer treatment strategy is herein described.

摘要

背景/目的：缺氧诱导因子-1α（HIF-1α）在细胞对缺氧的反应中起关键作用，而缺氧在异常血管生成和转移的诱导中起关键作用。了解通过HIF-1α调节性微小RNA调控血管生成的机制将有助于制定预防转移的策略。

材料与方法

我们通过评估微小RNA模拟物对HIF-1α表达的影响，对抑制HIF-1α的微小RNA进行了功能筛选，并通过机制研究确定miR-5586-5p为血管生成抑制剂。使用暴露于来自微小RNA转染的乳腺癌细胞的条件培养基中的人脐静脉内皮细胞（HUVEC）进行管形成试验，评估血管生成活性。通过向BALB/c裸鼠乳腺脂肪垫注射MDA-MB-231细胞建立原位异种移植小鼠，通过瘤内注射微小RNA检测miR-5586-5p的体内活性。

结果

关键促血管生成因子血管内皮生长因子A（VEGFA）和血管生成素样蛋白4（ANGPTL4）的表达受到miR-5586-5p的抑制。在由miR-5586-5p转染细胞制备的条件培养基中，人脐静脉内皮细胞的迁移和管形成减少。miR-5586-5p还抑制了肝素结合表皮生长因子样生长因子（HBEGF）以及在缺氧信号传导中发挥作用以诱导VEGFA和ANGPTL4表达的解整合素和金属蛋白酶17（ADAM17）的表达。HIF-1α、HBEGF和ADAM17被证实为miR-5586-5p抑制血管生成功能的直接靶点。乳腺癌患者肿瘤组织中miR-5586-5p的表达水平低于相邻正常组织。miR-5586-5p的表达与HIF-1α、HBEGF、ADAM17、VEGFA和ANGPTL4的表达呈负相关。在小鼠原位MDA-MB-231异种移植瘤中瘤内注射miR-5586-5p可抑制血管生成及随后的肿瘤生长。