中文摘要：

目的：采用微球体培养法富集原代乳腺癌干细胞,并探讨微球体形成与乳腺癌临床病理特征之间的相关性。方法：采用细胞悬浮培养法对45例乳腺癌组织来源的原代乳腺癌细胞进行微球体培养。采用FCM分析CD44＋/CD24low/-表型细胞所占百分率,采用实时荧光定量PCR（real-time fluorogenic quantitative-PCR,RFQ-PCR）检测干细胞相关基因Nanog、KLF4、OCT-4、SOX2和MDR1在微球体细胞中的表达,分析微球体形成与乳腺癌临床病理特征之间的关系。结果：细胞悬浮培养14d时,原代乳腺癌细胞微球体形成。乳腺癌组织来源的原代乳腺癌细胞微球体细胞中CD44＋/CD24low/-表型细胞所占百分率高于原代乳腺癌细胞（分别为24.71%和1.30%,P〈0.05）。原代乳腺癌细胞微球体形成与发病年龄、初潮年龄、绝经情况、组织学分级、孕激素受体（progestogen receptor,PR）状态、人表皮生长因子受体2（human epidermal growth factor receptor2,HER-2）状态和肿瘤复发明显相关（P〈0.05）。微球体形成与否与无病生存期（disease-free survival,DFS）和总生存期（overall survival,OS）无明显相关性（P〉0.05）。结论：细胞悬浮培养法可有效富集原代乳腺癌干细胞,微球体形成与部分临床病理特征存在相关性,可能是影响预后的因素之一。

英文摘要：

Objective： To explore the enrichment of breast cancer stem cells derived from primary breast cancer tissues by using a suspended culture system, and to analyze the relationship between mammosphere formaiton and the clinicopathological features of patients with breast cancer. Methods： Breast cancer cells were obtained from 45 patients with breast cancer and cultured in a suspended culture system to generate mammospheres. The percentage of breast cancer cells with a phenotype of CD44＋/CD24 low/– in mammosphere cells was assessed by flow cytometry （FCM）, and the expressions of Nanog, KLF4, OCT-4, SOX2 and MDR1 genes were detected by real-time fluorogenic quantitative-PCR （RFQ-PCR）. The relationship between mammosphere formation and the clinicopathological features of patients with breast cancer was analyzed. Results： The mammospheres were formed on the 14th day during the period of suspended culture. FCM result demonstrated that the percentage of breast cancer cells with a phenotype of CD44＋/CD24 low/– in mammosphere cells derived from primary breast cancer cells was significantly higher than that in the primary breast cancer cells （24.71% vs 1.30%,P 〈 0.05）. The mammosphere formation was positively related to the age of breast cancer onset, age at menarche, menstrual status, histological grading, progestogen receptor status, human epidermal growth factor receptor 2 （HER-2） status and cancer recurrence （P 〈 0.05）. However, the disease-free survival （DFS） and the overall survival （OS） were not associated with mammosphere formation （P 〉 0.05）. Conclusion： Primary breast cancer stem cells can be enriched by using a suspended culture system, and the mammosphere formation was positively associated with some clinicopathological characteristics, so it suggests that the mammosphere formation may become a potential predictive factor in the prognosis of breast cancer.