Effect of miR-335 on biological behavior of breast cancer MDA-MB-231 cells

伍家燕; 高月; 曾帆; 李海玉; 樊建军; 李韵; 张寒韬; 刘革力; 宋方洲

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Effect of miR-335 on biological behavior of breast cancer MDA-MB-231 cells

更新时间：2026-01-09

- Effect of miR-335 on biological behavior of breast cancer MDA-MB-231 cells
- Journal of Army Medical University Vol. 37, Issue 14, Pages: 1406-1411(2015)
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  CLC： R394.2;R730.23;R737.9
- Published：2015
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伍家燕,高月,曾帆,李海玉,樊建军,李韵,张寒韬,刘革力,宋方洲.miR-335对MDA-MB-231乳腺癌细胞生物学特性的影响[J].陆军军医大学学报(原第三军医大学学报),2015,37(14):1406-1411.

　Wu Jiayan,Gao Yue,Zeng Fan,Li Haiyu,Fan Jianjun,Li Yun,Zhang Hantao,Liu Geli,Song Fangzhou.Effect of miR-335 on biological behavior of breast cancer MDA-MB-231 cells[J].J Amry Med Univ (J Third Mil Med Univ),2015,37(14):1406-1411.
伍家燕,高月,曾帆,李海玉,樊建军,李韵,张寒韬,刘革力,宋方洲.miR-335对MDA-MB-231乳腺癌细胞生物学特性的影响[J].陆军军医大学学报(原第三军医大学学报),2015,37(14):1406-1411. DOI：

　Wu Jiayan,Gao Yue,Zeng Fan,Li Haiyu,Fan Jianjun,Li Yun,Zhang Hantao,Liu Geli,Song Fangzhou.Effect of miR-335 on biological behavior of breast cancer MDA-MB-231 cells[J].J Amry Med Univ (J Third Mil Med Univ),2015,37(14):1406-1411. DOI：

摘要

目的检测 microRNA-335（miR-335）在乳腺癌细胞系BT474、T47D、 MCF-7、SK-BR-3、 MDA-MB-231和正常乳腺细胞系MCF-10A中的表达以及在原发性乳腺癌和淋巴结转移性乳腺癌中的表达，进一步研究miR-335对MDA-MB-231细胞增殖、凋亡及迁移能力的影响。方法采用荧光实时定量 PCR 检测miR-335 在乳腺癌细胞系BT474、T47D、 MCF-7、SK-BR-3、 MDA-MB-231和正常乳腺细胞系MCF-10A中的表达，以及在原发性乳腺癌和淋巴结转移性乳腺癌中的表达。体外培养MDA-MB-231细胞并将其分3组，分别为miR-335组、阴性对照组和空白对照组。miR-335组转染miR-335模拟物 (has-miR-335 mimics)，阴性对照组转染阴性对照序列(negative control，NC)，空白对照组细胞不行任何转染。通过 MTT实验检测过表达miR-335对细胞增殖能力的影响；流式细胞仪检测过表达miR-335对细胞凋亡的影响；Transwell实验检测对细胞迁移能力的影响。Western blot 检测MMP-2和MMP-9蛋白的表达。结果各乳腺癌细胞系中miR-335表达水平均低于乳腺正常上皮细胞系MCF-10A， MDA-MB-231细胞中最低（P0.01）。在随机配对的10例原发性乳腺癌和10例淋巴结转移性乳腺癌标本中，有7例淋巴结转移性乳腺癌中MiR-335的表达低于对应的原发性乳腺癌（P0.05）。过表达miR-335对乳腺癌细胞 MDA-MB-231 增殖、凋亡无明显影响 (P0.05)，但能显著抑制细胞的迁移能力 (P0.05)。Western blot 结果显示：miR-335组MMP-2和MMP-9蛋白的表达量均低于阴性对照组和空白对照组。结论 miR-335 在高转移性乳腺癌细胞系和淋巴结转移性乳腺癌组织中明显低表达，过表达miR-335对MDA-MB-231细胞的增殖、凋亡无明显影响，但抑制MDA-MB-231细胞的迁移能力。miR-335 可能通过下调MMP2和MMP9的表达从而抑制 MDA-MB-231 细胞迁移。

Abstract

Objective To determine the expression of miR-335 in breast cancer cells (BT474

T47D

MCF-7

SK-BR-3

and MDA-MB-231) and normal breast cells (MCF-10A) and in primary breast cancer and lymph node metastatic breast cancer

and investigate the effect of miR-335 on the proliferation

apoptosis

and migration of MDA-MB-231 cells. Methods MDA-MB-231 cells were divided into 3 groups: a miR-335 group (transfected with miR-335 mimics)

a control group (transfected with negative control) and a blank group (non-transfected). The expression of miR-335 in normal breast cells and breast cancer cells was detected by real-time PCR. MTT assay was performed to observe the proliferation ability. The effect of over-expression of miR-335 on cell apoptosis was determined by flow cytometry. Transwell chamber assay was carried out to investigate the effect of miR-335 on migration of MDA-MB-231 cells. The expression of MMP-2 and MMP-9 was detected by Western blotting. Results The level of miR-335 in the breast cancer cells

especially

MDA-MB-231 cells

was lower than that in the normal breast cells (P0.01). In the tissues of randomly paired primary breast cancer (10 cases) and lymph node metastatic breast cancer (10 cases)

the expression of miR-335 in 7 cases of lymph node metastatic breast cancer was lower than that in the corresponding primary breast cancer (P0.05). Over-expression of miR-335 had no significant effect on the proliferation and apoptosis of MDA-MB-231 cells (P0.05)

but could significantly inhibit the cell migration (P0.05). Western blot results showed that the protein levels of MMP-2 and MMP-9 in the miR-335 group were lower than those in the control group and the blank group. Conclusion The expression of miR-335 is particularly reduced in the MDA-MB-231 cells and in the lymph node metastatic breast cancer tissues. The over-expression of miR-335 markedly inhibits the migration of MDA-MB-231 cells

but has no effect on the cell proliferation and apoptosis. MiR-335 inhibits the migration of MDA-MB-231 cells through down-regulating the levels of MMP-2 and MMP-9.

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