BMP9对人乳腺癌细胞MDA-MB-231骨转移的影响及可能机制

中国生物工程杂志

2012, Vol. 32

Issue (03): 7-13

研究报告

BMP9对人乳腺癌细胞MDA-MB-231骨转移的影响及可能机制

孙笑笑¹, 王科², 冯红蕾¹, 刘月红¹, 万绍恒¹, 罗进勇¹, 张彦¹

1. 重庆医科大学临床检验诊断学教育部重点实验室重庆 400016;
2. 重庆医科大学附属永川医院重庆 402160

Effects and Possible Mechanism of BMP9 on the Bone Metastasis of Human Breast Cancer Cells MDA-MB-231

SUN Xiao-xiao¹, WANG Ke², FENG Hong-lei¹, LIU Yue-hong¹, WAN Shao-heng¹, LUO Jin-yong¹, ZHANG Yan¹

1. The Key Laboratory of Laboratory Medical Diagnostics in the Ministry of Education, Chongqing Medical University, Chongqing 400016, China;
2. The Laboratory Department, Affiliated Yongchuan Hospital, Chongqing Medical University, Chongqing 402160, China

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摘要：

探讨BMP9对人乳腺癌MDA-MB-231细胞骨转移能力的影响及其可能的机制。扩增高滴度的BMP9表达腺病毒,感染MDA-MB-231细胞,制备表达BMP9的重组MDA-MB-231/ BMP9细胞,以此作为实验组;同时以含GFP的空载腺病毒感染该细胞为MDA-MB-231/GFP,联合MDA-MB-231共同作为对照组;RT-PCR及Western blot检测重组MDA-MB-231/ BMP9细胞中BMP9以及磷酸化Smad1(PSmad1)的表达;定量PCR及Western blot检测三组细胞中CTGFmRNA和蛋白水平的表达情况,最后结合X片运用免疫组织化学染色的方法检测三组标本中CTGF的表达。结果发现重组MDA-MB-231/ BMP9细胞中存在BMP9的表达;与对照组细胞相比,MDA-MB-231/ BMP9细胞中存在PSmad1的活化增强及CTGF的表达下调;X片发现实验组裸鼠胫骨溶骨性缺损减少;瘤体组织免疫组化发现实验组CTGF表达下调。所以BMP9可以在体内抑制乳腺癌MDA-MB-231细胞的骨转移并且这种抑制作用有可能是通过下调CTGF来实现的。

关键词： BMP9; 骨转移; CTGF; 乳腺癌

Abstract:

The purpose is to investigate the effects and possible mechanisms on BMP9 inhibiting the bone metastasis of human breast cancer MDA-MB-231 cells. High titer adenovirus vector expressing BMP9 was used to infect MDA-MB-231 cells. Experimental group is MDA-MB-231/BMP9 cells,control groups are both MDA-MB-231/GFP cells and MDA-MB-231 cells. RT-PCR and Western blot were used to detect the expression of BMP9 and PSmad1 in recombinant MDA-MB-231/BMP9 cells; Q-PCR and Western blot were used to detect the expression of CTGF in three groups cells; At last, X Light and Immunohistochemistry were used separately to detect the osteolytic lesions and the expression of CTGF in three groups cells. BMP9 was expressed in recombinant MDA-MB-231/BMP9 cells; the recombinant MDA-MB-231/BMP9 cells exhibited higher level of phosphorylated Smad1 and lower level of CTGF than the MDA-MB-231/GFP cells and MDA-MB-231 cells, but total Smad1 protein was similar in the three groups. In comparison with the control groups, the osteolytic lesions observed were significantly reduced in MDA-MB-231/BMP9 group; The removed tumor samples were detected by immunohistochemistry, MDA-MB-231/BMP9 samples analysed showed less CTGF staining than the control samples. Taken together, BMP9 can inhibit the bone metastasis of breast cancer cells MDA-MB-231 in vivo and the downregulation of CTGF is the possible mechanism.

Key words: BMP9 Bone metastasis CTGF Breast cancer

收稿日期: 2011-12-12 出版日期: 2012-03-25

ZTFLH:

R737.9

基金资助:

国家自然科学基金资助项目(81172017,30800658)

通讯作者: 张彦,zy2753@hotmail.com E-mail: zy2753@hotmail.com

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引用本文:

孙笑笑, 王科, 冯红蕾, 刘月红, 万绍恒, 罗进勇, 张彦. BMP9对人乳腺癌细胞MDA-MB-231骨转移的影响及可能机制[J]. 中国生物工程杂志, 2012, 32(03): 7-13.

SUN Xiao-xiao, WANG Ke, FENG Hong-lei, LIU Yue-hong, WAN Shao-heng, LUO Jin-yong, ZHANG Yan. Effects and Possible Mechanism of BMP9 on the Bone Metastasis of Human Breast Cancer Cells MDA-MB-231. China Biotechnology, 2012, 32(03): 7-13.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/ 或 https://manu60.magtech.com.cn/biotech/CN/Y2012/V32/I03/7

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