Establishment and characterization of six canine hepatocellular carcinoma cell lines (Transcriptional changes induced by drug treatment with toceranib or sorafenib)

Hepatocellular carcinoma (HCC) is the most common malignant liver tumor in dogs. Although surgical resection is a major treatment option for canine HCC, there are no distinct strategies for unresectable tumor subtypes or adjuvant chemotherapy for tumors with positive margins. We aimed to establish and characterize novel HCC cell lines from canine patients. The cellular morphology, general growth features and tumorigenicity of the established cell lines were evaluated. We also examined the sensitivity of the cell lines to multi-target tyrosine kinase inhibitors (TKIs). Toceranib, a veterinary TKI that targets vascular endothelial growth factor (VEGFR)/platelet-derived growth factor receptor (PDGFR)/c-kit, effectively inhibited the mitogen-activated protein kinase pathway and induced apoptosis. The established canine HCC cell lines showed greater sensitivity to toceranib than to sorafenib, a first-line treatment for human HCC targeting RAF/VEGFR/PDGFR. Sorafenib showed improved anti-tumor effects when co-treated with SCH772984, an extracellular signal-regulated kinase inhibitor. Our study suggests new therapeutic strategies for canine HCC, and these cell lines are valuable research materials for understanding HCC tumor biology in both humans and dogs. We established canine HCC cell lines from hepatic tumors of six canine patients.To investigate the transcriptional changes induced by drug treatment in the cell lines, the cells were treated with toceranib (15μM) or sorefenib (30μM) for 24hrs. The cells treated with vehicle (sorafenib-DMSO, toceranib-DPBS) were used as control. Gene Ontology Biological Processes (GO-BP) enrichment analysis was performed using data obtained from RNA-seq of 4 different cell lines.

肝细胞癌（Hepatocellular carcinoma，HCC）是犬类最常见的肝脏恶性肿瘤。尽管手术切除是犬类HCC的主要治疗手段，但针对不可切除的肿瘤亚型或切缘阳性肿瘤的辅助化疗尚无明确方案。
本研究旨在从犬类患者体内建立并鉴定新型HCC细胞系，并对所建立细胞系的细胞形态、基本生长特性及致瘤性进行评估。
本研究同时检测了这些细胞系对多靶点酪氨酸激酶抑制剂（multi-target tyrosine kinase inhibitors，TKIs）的敏感性。
托塞拉尼布（Toceranib）是一款靶向血管内皮生长因子受体（Vascular Endothelial Growth Factor Receptor，VEGFR）、血小板衍生生长因子受体（Platelet-derived Growth Factor Receptor，PDGFR）及c-kit的兽用TKIs，可有效抑制丝裂原活化蛋白激酶通路并诱导细胞凋亡。
所建立的犬类HCC细胞系对托塞拉尼布的敏感性高于索拉非尼（sorafenib）；索拉非尼是人类HCC的一线治疗药物，靶向RAF、VEGFR及PDGFR。
当与细胞外信号调节激酶抑制剂SCH772984联合给药时，索拉非尼的抗肿瘤效果得到增强。
本研究为犬类HCC提供了全新的治疗思路，且这些细胞系可为研究人类与犬类的HCC肿瘤生物学特性提供极具价值的实验材料。
本研究从6例犬类患者的肝脏肿瘤中成功建立了犬类HCC细胞系。
为探究药物处理后细胞系的转录组变化，本研究将细胞分别用15μM托塞拉尼布或30μM索拉非尼处理24小时。
以经溶媒处理的细胞（分别为索拉非尼所用溶媒二甲基亚砜DMSO、托塞拉尼布所用溶媒DPBS）作为对照组。
本研究基于4株不同细胞系的RNA测序（RNA-seq）数据，开展了基因本体论生物学过程（Gene Ontology Biological Processes，GO-BP）富集分析。