Effects of knocking down miR-221-3p expression on the stemness maintenance and angiogenesis in breast cancer cells under hypoxia conditions

ZHANG Qian; LI Qi; MA Nan; ZHANG Wei-qun

doi:10.11659/jjssx.08E024072

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Effects of knocking down miR-221-3p expression on the stemness maintenance and angiogenesis in breast cancer cells under hypoxia conditions

Basic Research | 更新时间：2025-06-17

- Effects of knocking down miR-221-3p expression on the stemness maintenance and angiogenesis in breast cancer cells under hypoxia conditions
- JOURNAL OF REGIONAL ANATOMY AND OPERATIVE SURGERY Vol. 34, Issue 6, Pages: 471-477(2025)
- 作者机构：
  
  新疆维吾尔自治区人民医院乳腺、甲状腺科，新疆乌鲁木齐 830000
- 作者简介：
- 基金信息：
- DOI：10.11659/jjssx.08E024072
  CLC： R737.9
- Received：20 August 2024，
  
  Published：25 June 2025
- 稿件说明：
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张倩,李琦,马楠等.低氧条件下敲低miR-221-3p表达对乳腺癌细胞干性维持及血管生成的影响[J].局解手术学杂志,2025,34(06):471-477.

ZHANG Qian,LI Qi,MA Nan,et al.Effects of knocking down miR-221-3p expression on the stemness maintenance and angiogenesis in breast cancer cells under hypoxia conditions[J].JOURNAL OF REGIONAL ANATOMY AND OPERATIVE SURGERY,2025,34(06):471-477.
张倩,李琦,马楠等.低氧条件下敲低miR-221-3p表达对乳腺癌细胞干性维持及血管生成的影响[J].局解手术学杂志,2025,34(06):471-477. DOI： 10.11659/jjssx.08E024072.

ZHANG Qian,LI Qi,MA Nan,et al.Effects of knocking down miR-221-3p expression on the stemness maintenance and angiogenesis in breast cancer cells under hypoxia conditions[J].JOURNAL OF REGIONAL ANATOMY AND OPERATIVE SURGERY,2025,34(06):471-477. DOI： 10.11659/jjssx.08E024072.

摘要

目的

探究低氧条件下微小RNA-221-3p（miR-221-3p）在乳腺癌细胞中的表达情况，以及敲低其表达对乳腺癌细胞干性维持与血管生成的影响。

方法

将人乳腺癌细胞系MCF-7分别在常氧条件和低氧条件下培养，实时荧光定量PCR（RT-qPCR）检测细胞中miR-221-3p表达。将MCF-7细胞分为常氧组（在常氧条件下培养MCF-7细胞）、低氧组（在低氧条件下培养MCF-7细胞）、低氧+miR-221-3p NC组（在低氧条件下培养MCF-7细胞并转染miR-221-3p NC）、低氧+miR-221-3p inhibitor组（在低氧条件下培养MCF-7细胞并转染miR-221-3p inhibitor）。CCK-8法检测各组细胞增殖活性；肿瘤球形成实验检测各组细胞肿瘤球数目；流式细胞术检测各组细胞中干性标志物乙醛脱氢酶1（ALDH1）标记的细胞比例；Western blot检测各组细胞中分化抗原簇蛋白133（CD133）、跨膜黏附分子（CD44）、性别决定区Y框蛋白2（SOX2）蛋白表达；体外管形成实验检测各组细胞上清液培养下的人脐静脉血管内皮细胞（HUVEC）管形成数目；Western blot检测各组细胞中低氧诱导因子-1α（HIF-1α）、血管内皮生长因子（VEGF）蛋白表达。

结果

与常氧组比较，低氧组细胞中miR-221-3p及CD133、CD44、SOX2、HIF-1α、VEGF蛋白表达水平显著上调（

0.05），细胞增殖活性、肿瘤球数目、ALDH1

细胞比例、HUVEC成管数目均显著增加/升高（

0.05）；与低氧+miR-221-3p NC组比较，低氧+miR-221-3p inhibitor组细胞中miR-221-3p及CD133、CD44、SOX2、HIF-1α、VEGF蛋白表达水平显著下调（

0.05），细胞增殖活性、肿瘤球数目、ALDH1

细胞比例、HUVEC成管数目均显著减少/降低（

0.05）。

结论

在低氧条件下，乳腺癌细胞中miR-221-3p表达上调，敲低miR-221-3p表达能够降低乳腺癌细胞干性及血管生成能力。

Abstract

Objective

To investigate the expression of microRNA-221-3p （miR-221-3p） in breast cancer cells under hypoxia conditions， and the effects of knocking down its expression on the stemness maintenance and angiogenesis in breast cancer cells.

Methods

Human breast cancer cell line MCF-7 were cultured under normoxic and hypoxic conditions， and the expression of miR-221-3p in cells was detected by quantitative real-time polymerase chain reaction （RT-qPCR）. MCF-7 cells were divided into normoxic group （MCF-7 cells were cultured under normoxic conditions）， hypoxic group （MCF-7 cells were cultured under hypoxic conditions）， hypoxia+miR-221-3p NC group （MCF-7 cells were cultured under hypoxic conditions and transfected with miR-221-3p NC）， hypoxia+miR-221-3p inhibitor group （MCF-7 cells were cultured under hypoxic conditions and transfected with miR-221-3p inhibitor）. The proliferation activity of cells in each group was detected by CCK-8 method； the number of cell tumor spheres in each group was detected by tumor sphere formation assay； the proportion of cells labeled with the stemness marker aldehyde dehydrogenase 1 （ALDH1） in each group was detected by flow cytometry； the expression of cluster of differentiation 133 （CD133）， cluster of differentiation 44 （CD44） and sex determination region Y-box protein 2 （SOX2） of cells in each group was detected by Western blot； the number of human umbilical vein endothelial cells （HUVEC） tubes formed by cell supernatant culture in each group was detected by in vitro tube formation experiment； the expression of hypoxia-inducible factor-1α （HIF-1α） and vascular endothelial growth factor （VEGF） of cells in each group was detected by Western blot.

Results

Compared with normoxic group， the expression levels of miR-221-3p，and CD133， CD44， SOX2， HIF-1α， VEGF protein of cells in hypoxic group were significantly up-regulated （

0.05）， and the cell proliferation activity， the number of tumor spheres， the proportion of ALDH1

cells and the number of HUVEC tubes were significantly increased （

0.05）. Compared with hypoxia+miR-221-3p NC group， the expression levels of miR-221-3p，and CD133， CD44， SOX2， HIF-1α， VEGF protein of cells in hypoxia+miR-221-3p inhibitor group were significantly down-regulated （

0.05）， and the cell proliferation activity， the number of tumor spheres， the proport

ion of ALDH1

cells， and the number of HUVEC were significantly decreased （

0.05）.

Conclusion

Under hypoxic conditions， the expression of miR-221-3p in breast cancer cells is up-regulated， and knocking down the expression of miR-221-3p can reduce the stemness and angiogenesis ability of breast cancer cells.

关键词

Keywords

references

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