Department of Laboratory Animal Science of Hebei Medical University, Heibei Key Laboratory of Laboratory Animals, Shijiazhuang, 050017, Hebei, China.
Institute of Traditional Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin State Key Laboratory of Modern Chinese Medicine, Tianjin, 300193, China.
Sci Rep. 2020 May 20;10(1):8329. doi: 10.1038/s41598-020-65338-w.
The gloomy outcome of liver cancer is mainly due to the high rates of metastasis and recurrence, even after curative resection for early stage liver cancer. Our study was conducted to find the animal model suitable for the study of liver cancer metastasis. In our study, two liver cancer cells were obtained from N-nitrosodiethylamine (DEN) and N-nitrosomorpholine (NMOR) induced rats, and they were cultivated, screened and cloning cultivated. Bionomics of cells was analyzed. The results show that 2 cells had different metastatic potentiality. They were named Wrh-f2 and Wrh-s2, and they have the characteristics of Hepatocellular carcinoma cells. The bionomics of 2 cells showed: (1) The chromosome karyotype analysis showed that the mode of Wrh-f2 was 80-83 and Wrh-s2 was 55-57; (2) AFP positive cytoplasmic staining was observed in Wrh-f2 and Wrh-s2. Cytokeratin (CK) 7 and CK8 positive staining was present in Wrh-f2. CK8 positive staining was present in Wrh-s2; (3) The numbers of Wrh-f2 and Wrh-s2 that passed through the Transwells were 98 ± 12 and 55 ± 15;(4) Wrh-f2 had the significant higher colony formation (78%) than Wrh-s2(8%) (P < 0.01). (5) The animal models generated solid tumours when 2 cells were inoculated to nude mouse and rat. And Wrh-f2 developed stable pulmonary metastasis. The established cell lines with different metastatic potential showed obvious advantages over liver cancer in mimicking the biological properties of malignant liver cancer tumors. It provided a suitable model for the mechanism of liver cancer metastasis in vivo and in vitro.
肝癌预后不佳主要是由于即使对早期肝癌进行根治性切除后，仍存在高转移和复发率。本研究旨在寻找适合研究肝癌转移的动物模型。本研究从 N-亚硝基二乙胺（DEN）和 N-亚硝基吗啉（NMOR）诱导的大鼠中获得了两种肝癌细胞，并对其进行了培养、筛选和克隆培养。分析了细胞的生物学特性。结果表明，这两种细胞具有不同的转移潜能，分别命名为 Wrh-f2 和 Wrh-s2，具有肝癌细胞的特征。两种细胞的生物学特性如下：（1）染色体核型分析显示，Wrh-f2 的模式为 80-83，Wrh-s2 的模式为 55-57；（2）Wrh-f2 和 Wrh-s2 的细胞质中均有 AFP 阳性染色，Wrh-f2 中 CK7 和 CK8 阳性染色，Wrh-s2 中 CK8 阳性染色；（3）穿过 Transwell 的 Wrh-f2 和 Wrh-s2 的数量分别为 98±12 和 55±15；（4）Wrh-f2 的集落形成率（78%）明显高于 Wrh-s2（8%）（P<0.01）。（5）当将两种细胞接种到裸鼠和大鼠体内时，均可形成实体瘤，且 Wrh-f2 可形成稳定的肺转移。建立的具有不同转移潜能的细胞系在模拟恶性肝癌肿瘤的生物学特性方面明显优于肝癌，为体内和体外肝癌转移机制提供了合适的模型。
