摘要
目的检测乳腺癌细胞株McF-7与MCF-7／ADR的Notchl表达、成球能力、干细胞池比例及干细胞内Notchl的表达，探讨Notchl在逆转乳腺癌化疗耐药中的意义。方法实时荧光定量逆转录一聚合酶链反应（RT—qPCa）及Westernblot法检测乳腺癌细胞株MCF-7、MCF-7／ADR及表型为ALDH1＋／CD44＋／CD24-乳腺癌干细胞中Notchl的表达；无血清培养检测MCF-7及MCF-7／ADR的成球能力；流式细胞仪检测MCF-7与MCF-7／ADR的干细胞比例并分选干细胞。结果MCF-7／ADR的Notch1的表达明显高于MCF-7（0．0748±0．0043比0．0461±0．0022，P〈0．05），微球体形成能力也明显高于MCF-7（P〈0．05）；流式细胞仪检测干细胞比例，MCF-7／ADR高于MCF-7（11．40％比1．89％）；分选出的干细胞Notchl的表达明显高于非干细胞（0．3096±0．0324比0．0428±0．0061，P〈0．05）。结论Notchl在乳腺癌及乳腺癌干细胞中高表达，干预Notchl可能可以逆转乳腺癌治疗的耐药。
Objective To investigate the role of Notchl in the chemotherapy process of breast cancer by studying the the efficiency of mammosphere formation, the ratio of stem cells in MCF-7 and MCF-7/ADR, and the expression status of Notchl in MCF-7, MCF-7/ADR and stem cells respectively. Methods Real time fluorescent quantitative reverse transcriptase polymerase chain reaction （RT-qPCR） and Western blotting were employed to detect the expression status of Notchl in breast cancer cell lines MCF-7, MCF-7/ADR and ALDH1 ＋/CD44 ＋/CD24- breast cancer stem cells （BCSCs）. Serum-free me- dium was available to assess the efficiency of mammosphere formation. Flow cytometry was used to define the ratio of BCSCs and select them in MCF-7 and MCF-7/ADR. Results The expression status of Notchl [ （0. 0748±0. 0043 ） vs （0. 0461±0. 0022 ）, P 〈 0.05 ] and the ability of mammosphere formation were significantly higher in MCF-7/ADR than in MCF-7 （P 〈 0. 05）. There was significant difference in the ra- tio of BCSCs between MCF-7/ADR and MCF-7 （ 11.40% vs 1.89% ）. Furthermore the expression score of Notchl in BCSCs was significantly higher than in non-BCSCs [ （0. 3096±0. 0324 ） vs （0. 0428 ±0. 0061 ）, P 〈 0.05 ]. Conclusion The high level expression of Notchl in breast cancer and BCSCs suggests regulating Notchl may play an important role in reversing chemotherapy resistance of breast cancer.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2012年第5期818-820,共3页
Chinese Journal of Experimental Surgery
基金
国家自然科学基金资助项目（30972935）
