Initial Investigation Of The Establishment Of Subcutaneous Transplantation Models Of Human Hepatocellular Carcinoma In SCID Mice And NOD-SCID Mice

Objective:1.To establish a subcutaneous transplantation model of human hepatocellular carcinoma in immune defect mice.2.To investigate the immune micro-environment in primary hepatocellular carcinoma and transplantation tumor in immune defect mice.Methods: Fresh tissues of human hepatocellular carcinoma were transplanted subcutaneously into SCID mice and NOD-SCID mice.The occurrence rate, volume of tumor and metastasis were observed.The HE stained transplantation tumor tissues was observed. The infiltration of CD3+,CD4+,CD8+ T-lymphocytes number of the transplantation tumor tissues in NOD-SCID mice and the expression of CD3,CD4,CD8,FoxP3,CD20,CD56,CD68 in 30 cases of tumor tissues,peri-tumor tissues and non- tumor tissues of human hepatocellular carcinoma were counted and compared by immunohistochemistry.Result:Tissues of human hepatocellular carcinoma were both transplanted subcutaneously into SCID mice and NOD-SCID mice successfully. The occurrence rate was 13.33%, and average volume of tumor was 0.73±0.17cm3 in SCID mice. The occurrence rate was 30.00%, and average volume of tumor was 1.09±0.75cm3 in NOD-SCID mice. The occurrence rate in NOD-SCID mice was higher than that in SCID mice (P<0.05)and there were no difference with average volume of tumor between them(P>0.05). No metastasis were found in other organs in two kinds of models. The HE staining analysis comfirmed the transplantation tumor were similar with human hepatocellular carcinoma. The number of CD3 +,CD4 +,CD8 + T-lymphocytes of transplantation tumor tissues in NOD-SCID mice were 10.82±4.58/Hp,5.76±2.42/Hp,3.72±2.20/Hp。The number of CD3 +,CD4 +,FoxP3+ cells were the most in the peri-tumor tissue,the secondary in the tumor tissues,and the least in the corresponding non- tumor tissues (P <0.05);the number of CD8 +, CD56+,CD68+ cells were the most in the peri-tumor tissues,the secondary in the corresponding non- tumor tissues,and the least in in the tumor tissues (P <0.05). The number of CD20 + were no difference in the tumor tissues,peri-tumor tissues and the corresponding non- tumor tissues(P>0.05).The CD3+,CD4+,CD8+,CD56+,CD68+ cells in tumor tissues mainly spread on the area of mesenchyma,and the FoxP3+ cells spread among the whole tumor tissue.The number of CD3 +,CD4 +,CD8 + T-lymphocytes of transplantation tumor tissues in NOD-SCID mice were less than them of human hepatocellular carcinoma tissues,but the rates of CD4+/CD3+ and CD8+/CD3+ were no difference between them.Conclusions: The study successfully established the subcutaneous transplantation models of human hepatocellular carcinoma in SCID mice and NOD-SCID mice. The two models remained the pathomorphism feature of human hepatocellular carcinoma.The NOD-SCID mice model was superior in occurrence rate but costly with SCID mice model.and imitated the immunomicroenvironment of human hepatocellular carcinoma.It has great value for the future study of immunotherapy of human hepatocellular carcinoma.In immunomicroenvironment of human hepatocellular carcinoma, immunocytes with killer fuction decreased,and immunocytes with inhibitive fuction increased. The NOD-SCID mice model imitated the immunomicroenvironment of human hepatocellular carcinoma.