Transcriptomic analysis of mouse hepatic progenitors expressing different level of the HRasG12V oncogene (in vivo and ex vivo).

Analysis was done in parrallel on tissue culture and on orthotopic tumor xenografts from primary (liver) and metastatic (peritoneum) site after separation of tumoral and stromal cells components. Overall design: mRNA profiles of BMEL cells transduces with an empty vector or expressing eitheir low (RasLOW) or high (RasHIGH) levels of the HRasG12V oncogene were generated by deep sequencing, in triplicate, using Illumina HiSeq2500. mRNA profiles of tumor and stromal cells after dissociation and isolation from liver xenograft or peritoneal metastasis from 4 independant mice were generated using Illumina HiSeq2500

本研究在分离肿瘤细胞与基质细胞组分后，针对组织培养样本，以及源自原发性（肝脏）与转移性（腹膜）位点的原位肿瘤异种移植模型开展平行分析。整体实验设计：通过空载体转导，或分别表达低水平（RasLOW）、高水平（RasHIGH）HRasG12V癌基因的BMEL细胞的mRNA表达谱，采用Illumina HiSeq2500平台进行三次生物学重复的深度测序获得。此外，从4只独立小鼠的肝脏异种移植瘤或腹膜转移瘤中解离并分离得到的肿瘤细胞与基质细胞的mRNA表达谱，同样通过Illumina HiSeq2500平台测序生成。