Effect of siRNA mediated down-regulation of AnnexinⅡ on human breast cancer cell line MDA-MB-435S

GUO Bian-qin; CHEN Ting-mei; WEN Yang-an

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Effect of siRNA mediated down-regulation of AnnexinⅡ on human breast cancer cell line MDA-MB-435S

更新时间：2025-01-15

- Effect of siRNA mediated down-regulation of AnnexinⅡ on human breast cancer cell line MDA-MB-435S
- Journal of Army Medical University Vol. 31, Issue 3, Pages: 223-226(2009)
- 作者机构：
  
  重庆医科大学：医学检验系临床血液教研室，临床检验诊断学省部共建教育部重点实验室，重庆市重点实验室，附属第一医院检验科
- 作者简介：
- 基金信息：
- DOI：
  CLC： R329.28;R394.3;R737.9
- Published：2009，
  
  Published Online：15 February 2009，
- 稿件说明：
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GUO BIAN-QIN, CHEN TING-MEI, WEN YANG-AN. Effect of siRNA mediated down-regulation of AnnexinⅡ on human breast cancer cell line MDA-MB-435S. [J]. Journal of army medical university, 2009, 31(3): 223-226.
DOI：

GUO BIAN-QIN, CHEN TING-MEI, WEN YANG-AN. Effect of siRNA mediated down-regulation of AnnexinⅡ on human breast cancer cell line MDA-MB-435S. [J]. Journal of army medical university, 2009, 31(3): 223-226. DOI：

摘要

目的探讨RNA干扰（RNA interference

RNAi）技术沉默AnnexinⅡ基因表达对人乳腺癌细胞MDA-MB-435S生物学特性的影响。方法体外化学合成AnnexinⅡ序列特异性双链小干扰RNA（siRNA），脂质体介导转染人乳腺癌细胞株MDA-MB-435S，采用荧光显微镜观察转染效率。收集siRNA干扰的细胞，RT-PCR、蛋白印迹和免疫细胞化学法分别检测AnnexinⅡ mRNA和蛋白的抑制水平，并采用MTT法、流式细胞术测定细胞增殖能力和细胞周期的变化，Millicell小室测定细胞侵袭、迁移能力的变化。结果 AnnexinⅡ-siRNA能有效抑制AnnexinⅡ mRNA和蛋白的表达(

0.05)；经siRNA干扰后的细胞，细胞增殖能力显著减低 (

0.05)；G

期细胞明显增加，S期细胞明显减少(

0.05)；细胞侵袭、迁移能力明显降低(

0.05)。结论采用siRNA干扰AnnexinⅡ mRNA和蛋白表达后，能有效逆转乳腺癌细胞的某些恶性生物学特点，表明AnnexinⅡ与乳腺癌的发生、发展及转移有一定的关系。

Abstract

Objective To study the effect of siRNA silencing of AnnexinⅡ on human breast cancer cell line MDA-MB-435S. Methods The chemically-synthetic siRNA duplexes targeting AnnexinⅡ was transiently transfected into MDA-MB-435S cells by lipofectamine 2000

and the transfection efficiency was observed by fluorescence microscopy. After siRNA interfering cells were harvested

the expression of Annexin Ⅱ was investigated by of RT-PCR

Western blot and immuocytoc

hemical method. Furthermore

the activity of cell proliferation and cell cycle were detected by MTT and flow cytometry respectively

and the abilities of cell invasion and metastasis was evaluated using millicell chambers. Results Annexin Ⅱ-siRNA effectively inhibited the expression of Annexin Ⅱ mRNA and protein (

0.05); After siRNA interfering

the cell proliferation was suppressed (

0.05); And cell cycle was arrested at G

phase with a significant decrease of cells in S phase (

0.05); The ability of cell invasive and metastasis was depressed (

0.05). Conclusion siRNA interfering to Annexin Ⅱ effectively reverses some of the malignant biological characters of human breast cancer cell line MDA-MB-435S

implying that Annexin Ⅱ might be involved in the progression and metastasis of breast cancer cells.

关键词

RNA干扰乳腺癌AnnexinⅡ基因细胞增殖细胞周期侵袭转移

Keywords

RNA interferencebreast cancerAnnexinⅡ genecell proliferationcell cycleinvasion and metastasis

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