体外培养乳腺癌肿瘤干细胞及其耐药相关性研究

A study on the culture of breast cancer stem cells in vitro and drug resistance

肿瘤干细胞(CSCs)是肿瘤组织中存在的少数具有自我更新和多向分化潜能的细胞亚群,具有较强的放化疗抵抗能力,是导致肿瘤复发和转移的罪魁祸首.因此,建立研究肿瘤干细胞的耐药模型十分重要.建立乳腺癌肿瘤干细胞的体外培养体系,MCF-7细胞在此条件下形成细胞球.采用碱性磷酸酶染色、流式分析、实时荧光定量PCR和蛋白印迹等方法对细胞球的干细胞特性进行验证.随后,通过阿霉素(一种常用的化疗药物)毒性实验表明,肿瘤干细胞具有更低的药物摄取效率和更高的药物抵抗能力.另外,肿瘤干细胞高表达ABCG2转运蛋白,后者是细胞产生耐药性的原因.

Cancer stem cells （CSCs） stand for a small population of cancer cells that have the potential of self-renewal and differentiation and lead to strong chemotherapy resistance,tumor recurrence and metastasis.Es-tablishing a stable model for the study of CSC drug resistance is necessary. Herein a culture approach of cancer stem cells was developed by using serum-free suspension culture system, and MCF-7 breast cancer cell sphe- roids were formed by using the method.Cancer stem cells in these spheroids were identified via alkaline phospha- tase staining, flow cytometer analysis, quantitative Real-time PCR and Western Blot.Subsequently, doxorubicin, a common used chemotherapeutic drug,was applied to these CSCs,which showed lower drug uptake efficiency and high level of resistance compared to normal cultured MCF-7 cells.Further study showed that the membrane trans-porting protein ABCG2, was highly expressed in CSCs, which also related to the chemotherapeutic drug resistance.