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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 To investigate the inhibitory effect of bone morphogenetic proteins 9 (BMP9) on osteolytic bone metastasis of human breast cancer MDA-MB-231 cells and its possible mechanism. Methods Human breast cancer MDA-MB-231 cells were divided into experimental group (infected with BMP9 adenovirus), blank control group (untreated) and GFP control group (infected with GFP adenovirus). RT- The transcriptional level of osteoprotegerin (OPG) mRNA was detected by Western blot. The expression of BMP9 and OPG in each group were detected. The BALB / c nude mice were divided into blank control group (blank control cells injected), GFP control group (injected GFP control cells) and experimental group (injected test group cells), each group was injected with tibia, 1 × 106 / (0.3μl · only), X-ray imaging analysis of nude mice in each group changes in osteolytic area, immunohistochemistry method to detect the expression of OPG protein in the tumor tissue of each group. Results The mRNA and protein expression levels of OPG in MDA-MB-231 cells were significantly higher than those in blank control group and GFP control group (P <0.05). Only the expression of BMP9 was detected in MDA-MB-231 cells. The diameter and osteolytic region of tibia in nude mice in experimental group were significantly lower than those in GFP control group and blank control group (P <0.05). The expression of OPG in experimental group was significantly higher than that in GFP control group (P <0.05) ). Conclusion BMP9 can inhibit osteolytic bone metastasis of MDA-MB-231 cells, which may play an important role by up-regulating the expression of OPG in OPG / RANKL / RANK system. This experiment laid the foundation for exploring the mechanism of BMP9 in tumor bone metastasis microenvironment.