乳腺癌干细胞培养鉴定及分化过程中微环境的影响  

Identification and differentiation of breast cancer stem cells under tumor microenvironment

作　　者：张书卿[1] 张博[1] 洪亮[1] 周永安[1] 赵亮[1] 

机构地区：[1]湖北理工学院附属医院(湖北省黄石市中心医院)普爱院区乳腺肿瘤外科,湖北省黄石市435000

出　　处：《中国组织工程研究》2015年第14期2155-2160,共6页Chinese Journal of Tissue Engineering Research

摘　　要：背景:乳腺癌干细胞会对乳腺癌的发生、转移等产生较大的影响。通过模拟肿瘤微环境,可以更好的对乳腺癌干细胞增殖与分化情况进行分析。目的:探讨肿瘤微环境对乳腺癌干细胞分化过程的影响。方法:对乳腺癌细胞及MCF-7细胞进行成纤维细胞上清液和无血清培养液PCM-2原代培养,观察乳腺癌细胞微球体的形成情况,采用MTT比色法检测乳腺癌细胞增殖能力,利用免疫细胞化学方法和RT-PCR方法检测乳腺癌干细胞标记物和上皮间质标记物的表达情况。结果与结论:无血清培养液PCM-2培养的原代细胞微球体直径显著大于成纤维细胞上清液。成纤维细胞上清液培养速度显著快于无血清培养液PCM-2。无血清培养液PCM-2培养第3天原代细胞中ALDH1表达率显著高于成纤维细胞上清液。与无血清培养液PCM-2培养比较,成纤维细胞上清液培养原代细胞中的ALDH1表达出现下调现象,而E-cadherin、Vimentin等上皮间质标记物表达则出现上调现象。在成纤维细胞上清液培养MCF-7细胞中的E-cadherin、VimenT in表达上调,而ALDH1、Oct-4等乳腺癌干细胞标记物表达下调。结果表明肿瘤微环境会对乳腺癌干细胞生存状态和分化过程产生一定的影响,成纤维细胞上清液中分泌的一些因子可以从一定程度上对乳腺癌干细胞样微球体的增殖与分化产生促进作用。BACKGROUND：Breast cancer stem cel s have a greater impact on the occurrence and metastasis of breast cancer. Under simulated tumor microenvironment, we can better analyze the proliferation and differentiation of breast cancer stem cel s. OBJECTIVE：To explore the tumor microenvironment effect on the differentiation of breast cancer stem cel s. METHODS：Breast cancer cel s and MCF-7 cel s were primarily cultured in fibroblast supernatant and serum-free PCM-2 medium, and formation of breast cancer cel s microspheres was observed. Proliferative ability of breast cancer cel s was detected using MTT colorimetry, and the surface markers of breast cancer stem cel s and epithelial-mesenchymal transition markers were measured using immunocytochemistry and RT-PCR methods. RESULTS AND CONCLUSION：The diameter of primary cel microspheres was larger in the serum-free PCM-2 medium than in the fibroblast supernatant, but the culture speed was faster in the fibroblast supernatant than the serum-free PCM-2 medium. At 3 days of primary culture, the expression of ALDH1 in primary cel s was greatly higher in the serum-free PCM-2 medium than in the fibroblast supernatant. However, the expressions of E-cadherin and vimentin were up-regulated in the fibroblast supernatant than in the serum-free PCM-2 medium. In addition, the expressions of E-cadherin and vimentin in MCF-7 cel s cultured in the fibroblast supernatant were up-regulated, while the expressions of ALDH1 and Oct-4 were downregulated. These findings indicate that the tumor environment has some certain effects on the growth and differentiation of breast cancer stem cel s, and some cytokines secreted from fibroblast supernatant can promote the proliferation and differentiation of breast cancer stem cel microspheres to some extent.

关 键 词：干细胞 肿瘤干细胞 乳腺癌 乳腺癌干细胞 肿瘤微环境 微球体 培养 

分 类 号：R394.2[医药卫生—医学遗传学]