Transcription factor TWIST-1 overexpression in acute myeloid leukemia cell line U937

Alterations of TWIST-1 expression are often seen in solid tumors and contribute to tumorigenesis and cancer progression. However, studies concerning its pathogenic role in leukemia are scarce. Here we show that TWIST-1 is a new candidate gene contributing to leukemogenesis of myeloid leukemia. We used array as one tool to determine gene expression profiles of control and TWIST-1-overexpressing U937 cells. U937 cell line was transduced with TWIST-1 or empty vector. GFP+ cells were sorted. Microarray was performed at GMINIX in Shanghai. Total RNA was isolated with TRIzol reagent (Invitrogen, Canada) and purified using RNeasy Mini Kit (Qiagen, German), including a DNase digestion treatment. RNA concentrations were determined by NanoDrop 2000 (Thermo, America). RNA was labeled using the GeneChip® WT Terminal Labeling and Controls Kit combined with the Ambion® WT Expression Kit. Labeled cRNA was hybridized to GeneChip® Human Transcriptome Array 2.0 arrays (Affymetrix, America).

TWIST-1 表达异常常见于实体瘤，并可促进肿瘤发生与癌症进展。然而，有关其在白血病中的致病作用的研究却较为匮乏。本研究证实，TWIST-1是一个参与髓系白血病发生的新型候选基因。本研究以基因芯片技术为手段，检测对照组与过表达TWIST-1的U937细胞的基因表达谱：将TWIST-1过表达载体或空载体转导至U937细胞系中，随后分选出GFP阳性细胞。基因芯片检测在上海GMINIX完成。总RNA采用TRIzol试剂（Invitrogen，加拿大）提取，并使用RNeasy迷你试剂盒（Qiagen，德国）进行纯化，同时辅以DNase消化处理。RNA浓度通过NanoDrop 2000分光光度计（Thermo，美国）测定。采用GeneChip® WT末端标记与对照试剂盒结合Ambion® WT表达试剂盒对RNA进行标记。将标记后的cRNA与GeneChip®人类转录组芯片2.0（Affymetrix，美国）进行杂交。