成骨细胞诱导乳腺癌侧群细胞获得骨模拟特性的研究  

Osteomimicry of side population cells induced by osteoblast in vitro

作　　者：杨晓宁[1] 周艳[1] 齐晓伟[1] 王明浩[1] 范林军[1] 姜军[1] 

机构地区：[1]第三军医大学西南医院乳腺疾病中心,重庆400038

出　　处：《第三军医大学学报》2013年第11期1102-1106,共5页Journal of Third Military Medical University

基　　金：重庆市自然科学基金(CSTC2009BB5173)~~

摘　　要：目的以乳腺癌细胞株MDA-MB231和成骨细胞株MC3T3-E1为对象,研究在共培养条件下乳腺癌侧群细胞向成骨细胞转化的能力。方法通过流式细胞技术分选出MDA-MB231乳腺癌侧群细胞,将其与成骨细胞株MC3T3-E1按照1∶1的细胞比例分别置于Transwell间接共培养体系的上、下小室中(实验组),设置相同密度的单纯乳腺癌侧群细胞为对照组,观察侧群细胞的形态变化。通过细胞免疫荧光技术及Western blot,比较2组侧群细胞中OPN、Runx2、Wnt-1表达水平的改变。结果实验组细胞分化明显,细胞免疫荧光技术检测实验组侧群细胞中OPN、Runx2、Wnt-1表达水平明显高于对照组。Western blot检测表明实验组侧群细胞中OPN(5.15±0.35)、Runx2(6.22±0.36)、Wnt-1(9.30±0.28)的蛋白表达水平均高于对照组OPN(1.00±0.15)、Runx2(1.00±0.14)、Wnt-1(1.00±0.13),差异具有统计学意义(P<0.05)。结论与成骨细胞株共培养后,乳腺癌侧群细胞获得成骨细胞相关生物学特性。Objective To investigate the differentiation of side population cells from breast cancer cell line MDA-MB231 to osteoblasts after co-cultured with osteoblastic cell line MC3T3-E1 in Transwell chamber. Methods Side population cells were obtained by flow cytometry from MDA-MB231 cells. And then the obtained side population cells and MC3T3-E1 cells were placed into the upper and lower chambers of Transwell co-culture system respectively at a ratio of 1 to 1. Side population cells at the same density placed in the both chambers served as control. The morphology and proliferation of the side population cells was observed by phase-contrast microscopy. The expression of OPN, Runx2 and Wnt-1 was detected by immunofluorescence staining and Western blotting. Results The side population cells were obviously differentiated in the co-culture system. Immunofluorescence staining indicated the expression of above 3 proteins was significantly stronger than in the control. Western blotting showed the same results （OPN： 5.15±0.35 vs 1.00±0.15, Runx2： 6.22±0.36 vs 1.00±0.14, Wnt-1： 9.30±0.28vs 1.00±0.13, P〈0.05）. Conclusion Our co-culture system induces side population cells from MDA-MB231 cells to obtain relative biological characteristics of osteoblasts.

关 键 词：乳腺癌 侧群细胞 MDA—MB231 成骨细胞MC3T3-E1 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学] R322.71[医药卫生—基础医学]