采用PDX模型探讨索拉非尼治疗肝细胞癌早期应答基因改变的研究

Investigation of early response of HCC related gene to sorafenib in PDX

利用人源肿瘤异种移植(patient-derived xenograft, PDX)模型探讨索拉非尼灌胃后,肝细胞癌(hepatocellularcarcinoma, HCC)早期应答基因表达的动态改变。方法取临床肝癌新鲜手术切除标本,在高度免疫缺陷模型小鼠NPG(NOD-Prkdcscid I12rg-/-)皮下接种,建立PDX模型,成功后分别留取给药前、给药后24 h和48 h的组织样本,提取肝组织总RNA。应用Fluidigm Biomark高通量基因分析系统进行96个基因的表达检测,据此对发生变化的基因功能和参与的信号通路进行分析c结果根据IFold Change 1>2.0进行差异基因筛选,与未给药组织比较,给药后24 h共有17个基因表达上调,21个基因表达下调;给药后48 h共有6个基因表达上调,19个基因表达下调。两个时间点共有的差异表达基因为20个,基因本体论(gene ontology, GO)分析显示,差异基因主要调控细胞周期及细胞迁移等功能;对KEGG信号通路分析显示,差异基因主要调控的信号通路包括细胞因子-细胞因子受体相互作用、黏着斑、癌症信号通路、前列腺癌及结直肠癌等信号通路。结论PDX模型研究显示,在早期出现变化的基因功能及信号通路均为调控细胞周期,抑制细胞迁移运动相关,推断索拉非尼在体内最早可能通过调控该类信号通路发挥抑制HCC作用。

Objective To investigate the early response of HCC related-genes with patient-derived xenograft(PDX)mice after sorafenib administration. Methods Fresh surgical resection specimens of HCC were taken and subcutaneously inoculated into highly immunodeficient model NPG mice to establish HCC PDX model. After successful administration, the tissue samples were taken at pre-administration, 24 h and 48 h after administration and the total RNA was extracted from liver tissue. The Fluidigm Biomark high-throughput gene analysis system was used to detect the expression of 96 genes. The functions of genes that changed and signaling pathways involved were analyzed. Results Compared with the control group, 17 genes were up-regulated and 21 genes were down-regulated in 24 h. While in 48 h, six genes were up-regulated, 19 genes were down-regulated and 20 genes were differentially expressed at two time points(|Fold Change| >2.0). GOs analysis showed that the differentially expressed genes mainly regulated cell cycles and cell migration. The KEGG results showed that the main signaling pathways regulated by different genes included cytokine-cytokine receptor interaction, adhesion plaque,cancer pathway, prostate cancer and colorectal cancer. Conclusions The gene regulations of PDX mice show that early changes in gene function and signaling pathways are related to regulation of cell cycle, inhibiting cell migration and movement, suggesting that Sorafenib may inhibit HCC by regulating these signaling pathways in vivo as early as possible.