动物造模,小鼠循环肝癌细胞模型 z-bio 2021-10-22 753

　　Objective: To establish a mouse liver cancer circulating tumor cell model using mouse liver cancer Hepa1-6 cells.

　　Method: 108 male C57BL/6 mice were used, divided into 3 groups according to body weight, 36 mice in each group. Each of the three groups was inoculated with 0.2 mL mouse hepatocarcinoma Hepa1-6 cell single cell suspension in each tail vein at the concentration of 1 x 106, 5 x 106 and 1 x 107 cells/mL. Each group was tested and blood was drawn on days 1, 5, 9, 13, 17 and 21 after vaccination. Flow cytometry is used to record the number and percentage of CTC in 20,000 nucleated cells, as well as their relative CTC inhibition rate and animal mortality. (2) Divide 80 male C57BL/6 mice into two groups according to their body weights, namely the model control group and the sorafenib tosylate group, each with 40 mice. The tail vein was inoculated with 0.2 mL of Hepa1-6 single cell suspension with a concentration of 5×106 cells/mL, and 21 times of solabettosulfonate (50 mg/kg) was continuously inoculated on the second day after inoculation. Blood was collected on the 3rd, 8th, 15th and 21st day of the drug for CTC detection.

　　Results: (1) After inoculating the cell suspension at a concentration of 1×106 cells/mL, the animal CTC ratios of the first 1, 5, 9, 13, 17 and 21 days were 25.1%, 18.1%, and 8.9%, respectively , 4.4%, 2.9%, 0.3%, no animal died; after inoculating a cell suspension with a concentration of 5 x 106 cells/mL, the first 1, 5, 9, 13% of the animal’s CTC. The 17th and 21st days are as follows: 40.4%, 35.4%, 15.4%, 9.0%, 6.6%, 4.1%, no animal death; the cell concentration is 1 x 107 cells/mL The percentage of animals on day 1 and day 5 After inoculation of the suspension, the CTC was 39.1% and 33.5%. Animal death occurred immediately after the inoculation, and all animals died 7 days after the inoculation. (2) The relative clearance rates of circulating tumor cells D3, D8, D15 and D21 after the administration of sorafenib tosylate are -7.5%, 4.6%, 55.3% and -94.5%, respectively, compared with the model control group. Significant difference (). P\u003c0.05 or P\u003c0.01).

　　Conclusion: Intravenous injection of mouse hepatocellular carcinoma Hepa1-6 suspension with a concentration of 5×106 cells/mL can establish a mouse hepatocellular carcinoma CTC model, which can be used to screen and evaluate drug cells that inhibit circulating tumors.