摘要
目的探讨骨形态发生蛋白9(bone morphogenetic proteins 9,BMP9)对人乳腺癌MDA-MB-231细胞溶骨性骨转移的抑制作用及其可能的机制。方法将人乳腺癌MDA-MB-231细胞分为试验组(感染BMP9腺病毒)、空白对照组(未感染)及GFP对照组(感染GFP腺病毒),RT-PCR法检测各组细胞中骨保护素(osteoprotegerin,OPG)基因mRNA的转录水平,Western blot法分别检测各组细胞中BMP9及OPG蛋白的表达水平。将BALB/c裸鼠分为空白对照组(注射空白对照细胞)、GFP对照组(注射GFP对照组细胞)及试验组(注射试验组细胞),每组5只,均经胫骨贴骨注射,1×106个/(0.3μl·只),X片成像技术分析各组裸鼠溶骨区域变化,免疫组化法检测各组裸鼠瘤体组织中OPG蛋白的表达。结果试验组MDA-MB-231细胞中OPG基因mRNA的转录水平及蛋白的表达水平均明显高于空白对照组和GFP对照组(P<0.05),仅试验组细胞中有BMP9的表达。试验组裸鼠胫骨瘤体直径及溶骨区域均明显小于GFP对照组和空白对照组(P<0.05),试验组裸鼠瘤体组织中OPG的表达量明显高于GFP对照组(P<0.05)。结论BMP9可抑制MDA-MB-231细胞溶骨性骨转移,可能是通过上调OPG/RANKL/RANK系统中OPG的表达而发挥作用。本实验为探讨BMP9在肿瘤骨转移微环境中的作用机制奠定了基础。
Objective o investigate the inhibitory effect of bone morphogenetic protein 9 （BMP9） on osteolytic bone metasta- sis of human breast cancer MDA-MB-231 cells as well as the possible mechanism. Methods MDA-MB-231 cells were divided into three groups. The cells in test group were infected with adenovirus containing BMP9, while those in GFP group with adenovirus containing GFP, and those in blank control group were uninfected, then determined for transcrip- tion level of OPG mRNA by RT-PCR, and for expressions of BMP9 and OPG by Western blot. Nude BALB / c mice were divided into blank control, GFP control and test groups, and injected with MDA-MB-231 cell suspension, in the cor- responding groups into shin bone, 0. 3 μl （1 ×106 cells） for each. The change in osteolytic regions of nude mice in vari- ous groups were observed by X ray imaging, while the expression of OPG by immunohistoehemical assay. Results Both the expression levels of OPG mRNA and protein in MDA-MB-231 cells in test group were significantly higher than those in blank control and GFP control groups （P 〈 0. 05）. BMP9 was only expressed in MDA-MB-231 cells in test group. Both the sizes of tumor and osteolytic region of mice in test group were significantly smaller than those in GFP control and blank control groups （P 〈 0. 05）. However, the OPG expression level in test group was significantly higher than that in GFP control group （P 〈 0. 05）. Conclusion BMP9 inhibited the osteolytic bone metastasis of MDA-MB-231 cells by a possible mechanism of upregulating the expression of OPG in OPG/RANKL/RANK system,which laid a foundation of investiga- tion on the action mechanism of BMP9 in microenvironment of bone metastasis of tumors.
出处
《中国生物制品学杂志》
CAS
CSCD
2014年第1期62-66,共5页
Chinese Journal of Biologicals
基金
国家自然科学基金资助项目(81172017)
重庆医科大学附属永川医院院内课题(YJQN20120023)
