论文部分内容阅读
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。
OBJECTIVE: To enrich primary breast cancer stem cells by microspheres culture and to explore the correlation between microsphere formation and clinicopathological features of breast cancer. Methods: 45 primary breast cancer cells derived from breast cancer cells were cultured in microspheres by cell suspension culture. FCM was used to analyze the percentages of CD44 + / CD24low / - phenotype cells and the expression levels of Nanog, KLF4, OCT-4, SOX2 and MDR1 were detected by real-time fluorogenic quantitative-PCR The expression of microspheres and the relationship between microsphere formation and clinicopathological features of breast cancer. Results: The primary breast cancer cells formed microspheres on the 14th day after suspension culture. The percentage of CD44 + / CD24low / - phenotype cells in breast cancer tissue derived primary breast cancer cells was significantly higher than that in primary breast cancer cells (24.71% and 1.30%, P <0.05, respectively). Primary breast cancer cell microspheres formation and onset age, menarche age, menopause, histological grade, progestogen receptor (PR) status, human epidermal growth factor receptor 2 (HER- 2) There was a significant correlation between tumor status and tumor recurrence (P <0.05). There was no significant correlation between microsphere formation and disease-free survival (DFS) and overall survival (OS) (P> 0.05). CONCLUSION: Cell suspension culture method can effectively enrich primary breast cancer stem cells. The formation of microspheres is correlated with some clinicopathological features and may be one of the factors that influence prognosis.