三维培养下脂肪细胞对乳腺癌MCF-7细胞迁移影响及其机制的初步研究

Adipocytes enhance the migration capacity of MCF-7 and their mechanisms under the 3D culture system

目的自体脂肪移植在乳腺癌患者乳房重建中的应用日益广泛,但移植的脂肪细胞与乳腺癌细胞发生哪些相互作用目前研究较少。本研究通过构建三维培养的乳腺癌细胞与脂肪细胞共培养模型,观察脂肪细胞对乳腺癌细胞迁移的影响及潜在分子机制。方法采用Matrigel胶对乳腺癌MCF-7细胞进行三维培养,观察细胞生长形态及方式;取成年女性腹部脂肪组织进行接种培养,通过Transwell小室建立乳腺癌MCF-7细胞与脂肪细胞共培养模型及MCF-7单独培养模型,比较MCF-7细胞迁移水平的变化;采用Affymetrix表达谱芯片分别构建共培养后及单独培养的MCF-7细胞表达谱,寻找差异表达基因。结果乳腺癌MCF-7细胞在Matrigel胶中生长呈现多细胞球形体;共培养12h后,共培养组MCF-7细胞每高倍镜视野迁移数为(267±48)个,单独培养组为(125±21)个,共培养后的MCF-7细胞迁移能力显著增强,t=5.421,P=0.005 9。表达谱芯片共筛出差异表达基因18个,其中表达上调基因15个,表达下调基因3个。结论脂肪细胞能够显著增加乳腺癌MCF-7细胞的迁移能力,推测可能与BCAS1、S100P、S100A10、S100A11及ANXA2基因有关。

OBJECTIVE Growing evidence has shown the efficacy of autologous fat graft in women with breast-con- serving surgery, however, the interactions between adipocytes and breast cancer cells are still limited. The aims of this study were to establish the co-culture system with human breast cancer cell line MCF-7 and adipocytes, and to investigate whether adipocytes affected the migration capacity of breast cancer cells. METHODS Firstly, to observe the growth of MCF-7 under the three-dimension model, MCF-7 was cultured using the Matrigel. Secondly, using Transwell chamber, MCF-7 ceils under the three-dimension model were seeded in the upper chamber and co-cultured with or without adipo- cytes in the lower chamber. Then, the migration capacity of MCF-7 was compared between the above two groups. Final- ly, microarray-based mRNA profiling was usecL to identify differentially expressed genes in these two groups. RESULTS Under the three-dimension culture system, MCF-7 presented as multicellular spheroids. Twelve hours after co-culturing with adipocytes, the counts of MCF-7 were 267＋48 under higher power lens, compared to 125±21 in the control group （t= 5. 421, P= 0. 005 9）. Eighteen genes were differentially expressed, of which 15 were up-regulated and 3 were down- regulated. CONCLUSION Adipocytes can promote the migration of MCF-7, and the effects are possibly associated with the genes of BCAS1, S100P, S100A10, S100A11 and ANXA2.