动物造模,循环肝癌细胞模型 z-bio 2021-08-20 722

　　Objective: To establish a mouse liver cancer circulating tumor cell model using mouse liver cancer Hepa 1-6 cells.

　　Method: 108 male C57BL/6 mice were used, and they were divided into 3 groups according to their body weight, with 36 mice in each group. Each of the three groups was inoculated with 0.2 mL of single cell suspension of mouse liver cancer Hepa 1-6 cells at a concentration of 1×106, 5×106, and 1×107 cells/mL in each tail vein. Each group was tested for blood sampling on the 1, 5, 9, 13, 17, and 21 days after vaccination. Flow cytometry was used to calculate the number and proportion of CTCs in 20,000 nucleated cells, and the relative CTCs inhibition rate, and record animal deaths Rate. ②80 male C57BL/6 mice were used and divided into 2 groups according to their body weight, namely the model control group and the sorafenib tosylate group, each with 40 mice. Each tail vein was inoculated with 0.2 mL of Hepa 1-6 cell single cell suspension at a concentration of 5×106 cells/mL, and sorafenib tosylate (50 mg/kg) was administered intragastrically on the second day after inoculation. For 21 consecutive days, blood was collected on the 3rd, 8th, 15th, and 21st day of administration for CTC detection.

　　Results: ① After inoculating the cell suspension with a concentration of 1×106 cells/mL, the proportion of animal CTCs on the first 1, 5, 9, 13, 17, and 21 days were: 25.1%, 18.1%, 8.9%, 4.4%, 2.9 %, 0.3%, no animal death occurred; after inoculation with a cell suspension at a concentration of 5×106 cells/mL, the proportion of animal CTCs on days 1, 5, 9, 13, 17, and 21 were: 40.4%, 35.4%, 15.4%, 9.0%, 6.6%, 4.1%, no animal death occurred; after inoculation with a cell suspension with a concentration of 1×107 cells/mL, the proportion of animal CTCs on the first and fifth days were: 39.1%, 33.5%, Animal death occurred immediately after the inoculation, and all animals died on the 7th day after the inoculation. ②D3, D8, D15, and D21 relative circulating tumor cell clearance rates during the administration of sorafenib tosylate were: -7.5%, 4.6%, 55.3%, -94.5%, respectively, which was significantly different from the model control group (P<0.05 or P<0.01).

　　Conclusion: Tail vein injection of Hepa 1-6 suspension of mouse liver cancer cells at a concentration of 5×106 cells/mL can establish a mouse liver cancer CTC model, which can be used for the screening and evaluation of drugs that inhibit circulating tumor cells.