模拟乳腺癌骨转移微环境中CGRP对成骨细胞OPG和RANKL表达影响  

Effects of calcitonin gene-related peptide on osteoprotegerin and RANKL expressions in osteoblast cells in bone metastasis microenvironment of breast cancer in vitro

作　　者：杨晨[1] 赵晖[1] 姚阳[1] 王智煜[1] 陈平[1] 

机构地区：[1]上海交通大学附属第六人民医院肿瘤内科,上海200233

出　　处：《中国肿瘤生物治疗杂志》2009年第3期243-247,共5页Chinese Journal of Cancer Biotherapy

基　　金：上海市市级医院慢性病综合防治项目(No.SHDC12007304)~~

摘　　要：目的:以乳腺癌细胞与成骨细胞共培养模拟乳腺癌骨转移微环境,观察在此微环境中降钙素基因相关肽(calci-tonin gene-related peptide,CGRP)对成骨细胞护骨素(osteoprotegerin,OPG)及细胞核因子κB受体活化因子配体(receptor acti-vator of nuclear factor-kappa B ligand,RANKL;又称破骨细胞分化因子)表达的影响。方法:将转移性乳腺癌细胞MDA-MB-231或MDA-MB-435与成骨细胞MG63共培养,建立模拟乳腺癌骨转移微环境。行CGRP(1×108mol/L)干预,应用RT-PCR和Western Blotting技术检测干预后OPG和RANKL在mRNA和蛋白水平表达的变化。结果:MG63与MDA-MB-231或MDA-MB-435共培养环境中,RANKL mRNA及蛋白水平升高,而OPG mRNA和蛋白水平表达下降;CGRP处理后,共培养环境中RANKL mRNA及蛋白水平降低,OPG mRNA和蛋白水平升高(均P<0.05)。结论:乳腺癌细胞能调节成骨细胞OPG/RANKL轴的表达,进而可能促进破骨细胞的活性,造成溶骨性破坏;CGRP干预可逆转此调节作用,在乳腺癌骨转移的治疗中有潜在应用价值。Objective ： To observe the effect of caleitonin gene-related peptide （CGRP） on the expression of osteoprotegerin （OPG） and receptor activator of nuclear factor-kappaB ligand （RANKL） in osteoblast cells through an in vitro breast cancer cell and osteoblast cell co-culture system. Methods： The metastatic breast cancer MDA-MB-231 or MDA- MB-435 cells were co-cultured with osteoblast MG63 cells to establish an in vitro mieroenvironment of bone metastasis of breast cancer. After treated with CGRP（1 × 10^8 mol/L） , OPG and RANKL mRNA and protein expressions in osteoblast MG63 cells were examined by RT-PCR and Western blotting. Results： Expression of RANKL in osteoblast MG63 cells was up-regulated at both mRNA and protein levels when osteoblast MG63 cells were co-cultured with breast cancer MDA- MB-231 or MDA-MB-435 cells, while those of OPG in osteoblast MG63 cells were both down-regulated （P 〈 0.05）. After treatment with CGRP, expressions of RANKL in osteoblast MG63 cells were down-regulated at both mRNA and protein levels, and the expressions of OPG mRNA and protein were both up-regulated （ P 〈 0.05 ）. Conclusion： Breast cancer MDA-MB-231 and MDA-MB-435 cells can promote osteolysis of osteoclast cells via regulating the expression of OPG/ RANKL axis in osteoblast cells. CGRP can reverse the osteolysis of osteoblast cells induced by breast cancer cells and may serve as a potential therapeutic agent for treatment of bone metastasis of breast cancer.

关 键 词：降钙素基因相关肽 乳腺癌细胞 成骨细胞 护骨素 破骨细胞分化因子 

分 类 号：R737.9[医药卫生—肿瘤] R730.2[医药卫生—临床医学]