乳腺癌细胞对成骨细胞增殖和分化功能的抑制作用  

Inhibitory effects of breast cancer cells on proliferation and differentiation of osteoblasts

作　　者：朱国英[1] 张燕燕[1] 顾淑珠[1] 陈晓[1] 

机构地区：[1]复旦大学放射医学研究所,上海200032

出　　处：《癌症》2009年第5期449-455,共7页Chinese Journal of Cancer

基　　金：上海市科委攻关计划(编号:064119512)~~

摘　　要：背景与目的:乳腺癌细胞对成骨细胞(osteoblast,OB)正常功能的影响作用尚不明确。本研究探讨人乳腺癌细胞MCF-7或MDA-MB-231条件培养基对OB正常功能的影响。方法:源于大鼠颅盖骨的原代OB与雌激素受体阴性(ER-)或阳性(ER+)的MDA-MB-231细胞或MCF-7细胞的条件培养基共同培养,MTT法观察条件培养基对OB增殖的影响,对硝基苯磷酸盐偶氮法观察条件培养基对OB碱性磷酸酶(alkaline phosphatase,ALP)活性的影响,茜素红S进行矿化结节染色并计算面积以观察条件培养基对OB矿化能力的影响。结果:MDA-MB-231细胞和MCF-7细胞条件培养基使OB发生纤维细胞形态改变,使细胞突起和突起连接均减少,并显著抑制OB的增殖。与培养基对照组比较,加入50%来自MDA-MB-231细胞或MCF-7细胞的条件培养基后1d、3d、5d和7d,OB的增殖抑制率分别为18.1%、13.0%、19.2%、19.3%和15.8%、20.8%、33.9%、28.7%,两组之间的差异均有统计学意义(P<0.01)。条件培养基可明显下调OB的ALP活性,并呈剂量-效应关系,50%的条件培养基使ALP活性分别下降31.9%和47.5%(P<0.01)。条件培养基对OB的矿化结节形成能力也呈明显抑制作用:加入50%MDA-MB-231细胞或MCF-7细胞的条件培养基培养后,OB形成矿化结节面积分别减少89%和74%(P<0.01)。结论:乳腺癌细胞可抑制OB的增殖能力和矿化形成能力,下调其AKP活性。Background and Objective= The effect of breast cancer cells on the normal function of osteoblasts remains unclear. This study was to investigate the effect of conditioned medium from two types of human breast cancer lines on osteoblastic proliferation, differentiation and mineralization. Methods： Fetal rat carvarial cells were treated with conditioned medium from MCF-7 estrogen-receptor（ER） positive, or MDA-MB-231 ER negatice, breast cancer cells. Proliferation was measured by MTT- assay, alkaline phosphatase （ALP） activity was assessed by the p-nitrophenyl phosphate （PNPP） method, and mineralized nodules were confirmed by alizarin red S （ARS） staining and the area of bone nodules was measured to observe the mineralization capacity. Results：When osteoblasts were treated with conditioned medium from MDA-MB-231 or MCF-7, the cells became long and spindle-like, assumed a fibroblastic morphology. Conditioned medium from breast cancer cells inhibited proliferation and differentiation of osteoblasts, compared with those cultured with vehicle control medium. The proliferation inhibition rates of culture with 50% conditioned medium from MDA-MB-231 and MCF-7 cells on osteoblasts were 18.1%, 13.0%, 19.2%, 19.3% and 15.8%, 20.8%, 33.9%, 28.7% on day 1, day 3, day 5, and day 7, respectively （P〈0.01）. Moreover, incubation with conditioned medium inhibited ALP activity and bone-nodule formation of osteoblasts in a dose-dependent manner. Addition of 50% conditioned medium from MDA- MB-231 and MCF-7 cells inhibited ALP activity by 31.9% and 47.5% （P〈 0.01）, and diminished the area of bone nodules by 89% and 74%, respectively, compared with the vehicle control group. Conclusion： Breast cancer cells inhibit proliferation, decrease ALP activity and disrupt the mineralization process of osteoblasts.

关 键 词：乳腺肿瘤 人乳腺癌细胞 成骨细胞 分化 增殖 矿化结节 碱性磷酸酶活性 

分 类 号：R737.9[医药卫生—肿瘤]