成骨前体细胞对乳腺癌细胞表达骨拟态特性及增殖的影响  

Effects of osteomimicry and proliferation of osteolytic breast cancer induced by osteoblastic progenitor cells in vitro

作　　者：马敏[1] 胡旭[1] 陈武桂 邱浩[1] 臧乐源[1] 胡建新[1] 周跃[1] 初同伟[1] 

机构地区：[1]第三军医大学新桥医院骨科,重庆400037

出　　处：《第三军医大学学报》2016年第9期954-958,共5页Journal of Third Military Medical University

基　　金：国家自然科学基金面上项目(81271979)~~

摘　　要：目的通过共培养体系探讨成骨前体细胞MC3T3-E1对MDA-MB-231乳腺癌细胞表达骨拟态特性及增殖的影响。方法实验组将MC3T3-E1及MDA-MB-231细胞采用transwell小室共培养,上室接种MC3T3-E1成骨细胞,下室接种相同数量的MDA-MB-231细胞。对照组上、下室均采用相同数量及密度的MDA-MB-231细胞。实时荧光定量PCR、Western blot法检测实验组及对照组骨钙蛋白(osteocalcin,OCN)、骨桥蛋白(osteopontin,OPN)、骨保护素(osteoprotegerin,OPG)的基因及蛋白的表达差异,采用CCK-8法检测各组增殖的差异,结晶紫染色法检测细胞集落形成差异。结果与对照组相比,实验组OCN、OPN、OPG的基因及蛋白表达均有明显的上调(P<0.05),MDA-MB-231细胞的增殖能力明显增高(P<0.01);实验组MDA-MB-231细胞集落形成数量及大小均优于对照组。结论短期共培养后MC3T3-E1成骨细胞可促进乳腺癌MDA-MB-231细胞表达骨拟态特性,并可促进其增殖及细胞集落形成。Objective To determine the effect of co-culturing with osteoblast cell line MC3T3-E1 on the osteomimicry and proliferation in the osteolytic breast cancer cell line MDA-MB-231. Methods MDAMB-231 cells were co-cultured with osteoblastic progenitor cells MC3T3-E1 in Transwell chamber in which MC3T3-E1 osteoblasts were inoculated in the upper chamber,while the same number of MDA-MB-231 cells in the lower chamber. In the control group,the same number and density of MDA-MB-231 cells were cultured in the upper and lower chambers,respectively. Real-time quantitative PCR and Western blotting were used to observe the expression of osteocalcin（ OCN）,osteopontin（ OPN）,and osteoprotegerin（ OPG） at mRNA and protein levels. Cell Counting Kit-8（ CCK-8） was employed to detect the proliferation in the 2 groups. Crystal violet staining was adopted to observe the difference of cell colony between the 2 groups. Results Compared with the control group,the expression levels of OPG,OPN and OCN were all increased at mRNA and protein levels in the experimental group（ P〈0. 05）. The proliferation of MDA-MB-231 was better in experimental group than the control group（ P〈0. 01）. Cell colony of MDA-MB-231 cells was better in the experimental group than the control group. Conclusion Short-term co-culturing with MC3T3-E1 cells promotes breast cancer MDA-MB-231 cells to form osteomimicry,and enhances the cell proliferation and colony formation at the same time.

关 键 词：成骨前体细胞 乳腺癌 骨拟态 MDA-MB-231 

分 类 号：R322.71[医药卫生—人体解剖和组织胚胎学] R730.23[医药卫生—基础医学]