摘要
目的利用流式细胞仪(FACS)细胞分选技术从乳腺癌细胞株MCF-7中分选乳腺癌干细胞,并利用基因芯片进行其miRNAs表达谱分析。方法利用已知的乳腺癌干细胞的表面分子标志(CD44+ESA+CD24-/low),以相应的荧光标记抗体对细胞加以标记,FACS细胞分选获得乳腺癌干细胞,NOD-SCID移植瘤实验进行干细胞功能鉴定;分别提取细胞总RNA以及小分子RNA,经荧光标记后与miRNAs基因芯片杂交,获得乳腺癌干细胞miRNAs表达谱。结果从MCF-7中分选到约1%的CD44+ESA+CD24-/low细胞,NOD-SCID移植瘤实验表明其具有干细胞特性,基因芯片杂交获得20个乳腺癌干细胞相关miRNAs,其中4个为低表达,16个为高表达。结论乳腺癌干细胞具有自身特征性miRNAs,为进一步从干细胞层面研究乳腺癌发病机制及靶向治疗打下基础。
Objectives To isolate breast cancer stem cells from MCF-7, an established breast cancer cell line, by FACS. And to analyse miRNAs expression profiles of the cancer stem cells by using microarray. Methods To isolate breast cancer stem cells of MCF-7, an established breast cancer cell line, on the basis of cell- surface markers using fluorescence-activated cell sorting （FACS） and to identify a subpopulation （ ESA＋ CD44 ＋ CD24 -/low）. The tumor-initiating capability of the ESA＋ CD44 ＋ CD24 -/low population was tested by transplantation into NOD/SCID mice. Next, using a microchip containing oligonucleotides corresponding to 517 miRNAs from human, mouse and rat genomes,miRNA gene expression profiles of both breast cancer stem cells and MCF- 7 were oblained respectively. Results CD44＋ESA＋CD24-/low cells, accounting for 1% of MCF-7 cells,were successfully isolated. The transplantation experiment demonstrated the properties that define the isolated subpopulation as stem cells. The microarray experiment showed 20 breast cancer stem cell-correlative miRNAs, 4 high and 16 low in breast cancer stem cell. Conclusion Breast cancer stem cell has a set of specific miRNAs, providing experimental basis for further research on mechanism of breast cancer through the roles of stem cell.
出处
《重庆医学》
CAS
CSCD
2007年第13期1280-1282,共3页
Chongqing medicine
基金
军队"十一五"计划科研基金资助攻关项目(06G069)
中国博士后科学基金资助项目(2005038478)
