miR-155 通过靶向 SOCS1 和 MMP16 促进乳腺癌细胞的增殖和迁移。

MiR-155 promotes the proliferation and migration of breast cancer cells via targeting SOCS1 and MMP16.

机构信息

Department of Thyroid Gland and Breast Surgery, the First Affiliated Hospital of Wenzhou Medical University, Wenzhou, China.

出版信息

Eur Rev Med Pharmacol Sci. 2018 Nov;22(21):7323-7332. doi: 10.26355/eurrev_201811_16269.

DOI:10.26355/eurrev_201811_16269

PMID:30468477

Abstract

OBJECTIVE

The aim of this study was to investigate the effect of miR-155 on the proliferation and migration of breast cancer cells, and to explore the underlying mechanism.

MATERIALS AND METHODS

The breast cancer cell line MDA-MB-231 was transfected with miR-155 mimics, inhibitor or negative control, respectively. The expression level of miR-155 was detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). Subsequently, the proliferation of MDA-MB-231 cells was detected by multi-cellular tumor spheroid (MTS) and colony formation assay. Cell migration was examined by transwell assay and scratch test. In addition, qRT-PCR was performed to analyze the expression of matrix metallopeptidase 16 (MMP16) after miR-155 mimics or inhibitor transfection in MDA-MB-231 cells. Meanwhile, Western blot was used to evaluate the protein expression levels of suppressor of cytokine signaling 1 (SOCS1) and MMP16 after miR-155 mimics or inhibitor transfection.

RESULTS

QRT-PCR results showed that miR-155 mimics significantly increased the expression of miR-155 in MDA-MB-231 cells, whereas miR-155 inhibitor markedly decreased miR-155 expression (p < 0.05). Meanwhile, MTS and colony formation assay indicated that the proliferation of MDA-MB-231 cells was remarkably increased after miR-155 mimics transfection. However, miR-155 inhibitor transfection exhibited the opposite result in cell proliferation (p < 0.05). Moreover, miR-155 overexpression significantly increased the migration of MDA-MB-231 cells (p < 0.05). Western blot further confirmed that miR-155 overexpression down-regulated the expression level of target protein SOCS1 and upregulated the expression level of MMP16.

CONCLUSIONS

We found that miR-155 significantly enhanced the proliferation and migration of MDA-MB-231 cells, which might serve as an oncogene in breast cancer. Therefore, it is preliminarily believed that miR-155 plays an important role in the proliferation and migration of breast cancer cells via down-regulating the expression of SOCS1 and up-regulating the expression of MMP16.

摘要

目的

本研究旨在探讨 miR-155 对乳腺癌细胞增殖和迁移的影响，并探讨其潜在机制。

材料和方法

分别用 miR-155 模拟物、抑制剂或阴性对照转染乳腺癌细胞系 MDA-MB-231。采用实时定量聚合酶链反应 (qRT-PCR) 检测 miR-155 的表达水平。随后，通过多细胞肿瘤球体 (MTS) 和集落形成实验检测 MDA-MB-231 细胞的增殖。通过 Transwell 测定和划痕试验检测细胞迁移。此外，在 MDA-MB-231 细胞中转染 miR-155 模拟物或抑制剂后，通过 qRT-PCR 分析基质金属蛋白酶 16 (MMP16) 的表达。同时，Western blot 用于评估 miR-155 模拟物或抑制剂转染后 MDA-MB-231 细胞中 SOCS1 和 MMP16 的蛋白表达水平。

结果

qRT-PCR 结果显示，miR-155 模拟物显著增加 MDA-MB-231 细胞中 miR-155 的表达，而 miR-155 抑制剂则显著降低 miR-155 的表达 (p<0.05)。同时，MTS 和集落形成实验表明，miR-155 模拟物转染后 MDA-MB-231 细胞的增殖显著增加。然而，miR-155 抑制剂转染则导致细胞增殖相反的结果 (p<0.05)。此外，miR-155 过表达显著增加 MDA-MB-231 细胞的迁移 (p<0.05)。Western blot 进一步证实，miR-155 过表达下调了靶蛋白 SOCS1 的表达水平，并上调了 MMP16 的表达水平。