CAJ | 학술논문

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、VimenTin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。
배경：유선암간세포회대유선암적발생、전이등산생교대적영향。통과모의종류미배경，가이경호적대유선암간세포증식여분화정황진행분석。목적：탐토종류미배경대유선암간세포분화과정적영향。방법：대유선암세포급MCF-7세포진행성섬유세포상청액화무혈청배양액PCM-2원대배양，관찰유선암세포미구체적형성정황，채용 MTT 비색법검측유선암세포증식능력，이용면역세포화학방법화 RT-PCR 방법검측유선암간세포표기물화상피간질표기물적표체정황。결과여결론：무혈청배양액PCM-2배양적원대세포미구체직경현저대우성섬유세포상청액。성섬유세포상청액배양속도현저쾌우무혈청배양액PCM-2。무혈청배양액PCM-2배양제3천원대세포중ALDH1표체솔현저고우성섬유세포상청액。여무혈청배양액PCM-2배양비교，성섬유세포상청액배양원대세포중적ALDH1표체출현하조현상，이E-cadherin、Vimentin등상피간질표기물표체칙출현상조현상。재성섬유세포상청액배양MCF-7세포중적E-cadherin、VimenTin표체상조，이ALDH1、Oct-4등유선암간세포표기물표체하조。결과표명종류미배경회대유선암간세포생존상태화분화과정산생일정적영향，성섬유세포상청액중분비적일사인자가이종일정정도상대유선암간세포양미구체적증식여분화산생촉진작용。
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.