动物造模,循环肝癌细胞模型 z-bio 2021-05-08 1028

　　OBJECTIVE: To establish a mouse circulating tumor cell (CTC) model of hepatocarcinoma using mouse 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 a concentration of 1 x 106, 5 x 106, and 1 x 107 cells/mL. Blood sampling tests were performed on each group on 1, 5, 9, 13, 17 and 21 days after vaccination. Flow cytometry is used to record the number and percentage of CTC in 20,000 nucleated cells, its relative CTC inhibition rate and animal mortality. (2) According to body weight, 80 male C57BL/6 mice were divided into two groups: model control group and sorafenib tosylate group, each with 40 mice. Each tail vein was inoculated with 0.2 mL of Hepa1-6 single cell suspension with a concentration of 5×106 cells/mL, and 21 consecutive injections of sorafenib tosylate (50 mg/kg) were inoculated after the next day. Then, on the 3rd, 8th, 15th and 21st days of administration, blood was collected for CTC detection.

　　Results: (1) After inoculating the cell suspension at a concentration of 1×106 cells/mL, the ratio of animal CTC on the first 1, 5, 9, 13, 17 and 21 days were 25.1%, 18.1% and 8.9, respectively. Yes. 4.4%, 2.9%, 0.3%, no animal deaths; the first 1, 5, 9, 13, 17 animals after inoculation with a cell suspension at a concentration of 5 x 106 cells/mL CTC. Day 21: 40.4%, 35.4%, 15.4%, 9.0%, 6.6%, 4.1%, no animal death; CTC rate of animals after inoculation with a cell suspension at a concentration of 1 x 107 cells/mL was 39.1% and 33.5 5% of the virus was observed on the 1st and 5th days, the animals died immediately after the inoculation, and all the animals died on the 7th day after the inoculation. (2) The relative circulating tumor cell clearance rates of D3, D8, D15 and D21 after sorafenib tosylate administration were -7.5%, 4.6%, 55.3% and -94.5%, compared with the model control group. Significant differences().

　　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.