Establishment of a tumor-bear mouse model carrying human HepG2 cells and exploration of immune tolerance

doi:10.16098/j.issn.0529-1356.2015.04.011

Abstract

Abstract:

Objective To establish anin vivo human hepatoma model carried by mice with normal immune background and to investigate the effect of immune tolerance. Methods We firstly generated green fluorescent protein (GFP) expressed HepG2 cells and intrauterine injected these cells into the peritoneal cavities of E16.5 day mouse fetus. Then we observed the change in newborn mice with bearing cancer and detect the human alpha fetoprotein (AFP) expression in mouse serum by elisa analysis. We subcutaneously injected the HepG2 cells into the adult mice and detected the lymphocyte subgroup ratio of spleen cells by flow cytometry. Results GFP-HepG2 cells were grown in fetus liver and secreted human AFP. After injection of GFP-HepG2 cells in fetus liver, some adult mice developed subcutaneous tumors. Flow cytometry analysis indicated that immune system of tumor-bear mice remained weak attack to human live cancer cells. Conclusion The tumor-bear model carrying human HepG2 cells may be established by injecting the GFP-HepG2 cells into the peritoneal cavities of mouse fetus, parts of which acquire the ability of immune tolerance.

Key words: Liver cancer, Bearing cancer, Immune tolerance, Flow cytometry, Mouse

LIU Chun-jia ZHANG Xing-yu SHAN Zhi-yan WU Yan-shuang LEI Lei*. Establishment of a tumor-bear mouse model carrying human HepG2 cells and exploration of immune tolerance[J]. AAS, 2015, 46(4): 509-513.

References

［1］Kluger MD, Salceda JA, Laurent A, et al. Liver resection for hepatocellular carcinoma in 313 Western Patients: tumor biology and underlying liver rather than tumor size drive prognosis［J］. J Hepatol, 2015, 62(5): 1131-1140.
［2］Okamura Y, Ashida R, Ito T, et al. The tumor marker score is an independent predictor of survival in patients with recurrent hepatocellular carcinoma［J］. Surg Today, 2014, 20.
［3］Ray S, Murmu N, Adhikari J, et al. Inhibition of HepG2 hepatic cancer cell growth and CCl4 induced liver cytotoxicity in Swiss albino mice by Mahua extract［J］. J Environ Pathol Toxicol Oncol, 2014, 33(4):295-314.
［4］Uehara T, Pogribny IP, Rusyn I. The DEN and CCl4-Induced Mouse Model of Fibrosis and Inflammation-Associated Hepatocellular Carcinoma［J］. Curr Protoc Pharmacol, 2014, 66(2):14. 30. 1-14. 30. 10. 
［5］Scaiewicz V, Nahmias A, Chung RT, et al. CCAAT/enhancer-binding protein homologous (CHOP) protein promotes carcinogenesis in the DEN-induced hepatocellular carcinoma model［J］. PLoS One, 2013, 8(12):e81065. 
［6］Troiani T, Schettino C, Martinelli E, et al. The use of xenograft models for the selection of cancer treatments with the EGFR as an example［J］. Crit Rev Oncol Hematol, 2008, 65(3):200-211.
［7］Chen Y, Ramjiawan RR, Reiberger T, et al. CXCR4 inhibition in tumor microenvironment facilitates anti-PD-1 immunotherapy in sorafenib-treated HCC in mice［J］. Hepatology, 2014, 20. 
［8］Xu YY, Chen L. Establishment of mouse liver cancer model［J］. Chinese Journal of Clinical and Experimental Pathology, 2011, 27 (4 ): 405-408. (in Chinese)
徐玉音,陈莉. 肝癌动物模型建立的方法［J］.临床与实验病理学杂志， 2 0 1 1， 2 7 (4 )： 405-408.
［9］Hanahan D, Weinberg RA. Hall marks of cancer: the next generation［J］. Cell, 2011, 144 (6 ): 646-674.
［10］Yi HF, Tang JM. Recent advances of dendritic tumour-cell fusion in antitumor immunity study［J］. Acta Anatomica Sinica, 2003, 34(6):665-666. (in Chinese)
伊焕发, 唐军民.树突状细胞-肿瘤细胞之融合细胞在抗肿瘤免疫中的研究进展［J］.解剖学报，2003，34(6):665-666. 
［11］Zhu G, Hong L, Wang S, et al. Comparison of two kinds of orthotopic xenograft models for human ovarian cancer［J］. Eur J Gynaecol Oncol, 2014, 35(6):724-727.
［12］Heijstek MW, Kranenburg O, Borel Rinkes IH. Mouse models of colorectal cancer and liver metastases［J］. Dig Surg, 2005, 22: 16-25.
［13］Li X, Shan ZY, Wu YSh, et al. Preliminary study of generation of human induced pluripotent stem cells from humun foreskin fibroblasts［J］. Acta Anatomica Sinica, 2012, 43(05):601-607. (in Chinese)
李雪，单智焱，吴嫣爽，等.人包皮成纤维细胞来源的多能性干细胞的建立［J］. 解剖学报，2012，43（05）：601-607.
［14］Duyang EC, Wu CH, Walton C, et al. Transplantion of human hepatocytes into tolerized genetically immunocompetewt rats［J］. World J Gastoenterol, 2001, 7(3):324-330.

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