基于基因芯片的乳腺癌干细胞miRNAs检测分析

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.

Analysis of miRNAs expression profiles of breast cancer stem cells by using microarray

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.