Whole genome expression microarray of cancer stem cells isolated from HT29 and HCT116 Human colorectal cancer cell lines

In the present study we have isolated and characterized cancer stem cells and non-cancer stem cells (bulk tumor cells) from high grade human colorectal cancer cell line HCT116 and low grade human colorectal cancer cell line HT29. For this study, cancer stem cells and non-cancer stem cells (bulk tumor cells) were isolated from HCT116 and HT29 human colorectal cancer cell line. For isolating cancer stem cells by FACS, CD44 and CD166 tagged with V450 and PE respectively were used. CD44+CD166+ was the cancer stem cell population and CD44-CD166- was designated as the non-cancer stem cell (bulk tumor cells) population for this study. RNA was isolated from the isolated cell populations and microarray was done to study the whole genome transcriptomic changes. 4 x 44k Whole genome expression arrays of FACS sorted HT29 cancer stem cells (n=2) and HT29 non-cancer stem cells (n=2) using total RNA obtained from FACS sorted cells. 4 x 44k Whole genome expression arrays of FACS sorted HCT116 cancer stem cells (n=2) and HCT116 non-cancer stem cells (n=2) using total RNA obtained from FACS sorted cells.

本研究从高分级人结直肠癌细胞系HCT116与低分级人结直肠癌细胞系HT29中分离并鉴定了癌症干细胞（cancer stem cells）与非癌症干细胞（bulk tumor cells，实体瘤细胞）。本研究中，我们从上述两种细胞系中分离得到目标细胞群体。为通过荧光激活细胞分选术（fluorescence-activated cell sorting, FACS）分离癌症干细胞，本研究分别使用标记V450的CD44与标记PE的CD166作为分选标志物；其中CD44⁺CD166⁺细胞群被定义为癌症干细胞群，CD44⁻CD166⁻细胞群则被定义为非癌症干细胞（bulk tumor cells）群。我们从分选得到的细胞群体中提取总RNA，并通过全基因组表达芯片分析全基因组转录组变化。本研究采用4×44k全基因组表达芯片，对经荧光激活细胞分选得到的HT29癌症干细胞（n=2）与HT29非癌症干细胞（n=2）的总RNA进行全基因组表达谱检测；同时采用相同芯片平台，对经荧光激活细胞分选得到的HCT116癌症干细胞（n=2）与HCT116非癌症干细胞（n=2）的总RNA进行全基因组表达谱检测。