【摘要】：目的采用小鼠肝癌Hepa 1-6細胞建立小鼠肝癌循環(huán)腫瘤細胞(CTCs)模型。方法采用雄性C57BL/6小鼠108只,按照體重分為3組,每組36只。3個組分別每只尾靜脈接種0.2 m L的濃度為1×106、5×106和1×107個/m L的小鼠肝癌Hepa 1-6細胞單細胞懸液。各組分別于接種后第1、5、9、13、17、21天檢測采血,采用流式細胞術(shù)檢測,計算2萬個有核細胞中CTCs數(shù)目及比例、相對CTCs抑制率,記錄動物死亡率。(2)采用雄性C57BL/6小鼠80只,按體重分為2組,分別為模型對照組、甲苯磺酸索拉菲尼組,每組40只。每只尾靜脈接種0.2m L的濃度為5×106個/m L的Hepa 1-6細胞單細胞懸液,接種后第2天開始灌胃給予甲苯磺酸索拉菲尼(50 mg/kg),連續(xù)21 d,并于給藥第3、8、15、21天采血進行CTC檢測。結(jié)果 (1)接種濃度為1×106個/m L的細胞懸液后,第1、5、9、13、17、21天的動物CTCs比例為:25.1%、18.1%、8.9%、4.4%、2.9%、0.3%,未出現(xiàn)動物死亡;接種濃度為5×106個/m L的細胞懸液后,第1、5、9、13、17、21天的動物CTCs比例為:40.4%、35.4%、15.4%、9.0%、6.6%、4.1%,未出現(xiàn)動物死亡;接種濃度為1×107個/m L的細胞懸液后,第1、5天的動物CTCs比例為:39.1%、33.5%,在接種完畢之后立即出現(xiàn)動物死亡,且所有動物在接種后第7天全部死亡。(2)甲苯磺酸索拉菲尼給藥期間D3、D8、D15、D21相對循環(huán)腫瘤細胞清除率分別為:-7.5%、4.6%、55.3%、-94.5%,與模型對照組比較差異有顯著性(P0.05或P0.01)。結(jié)論采用濃度為5×106個/m L小鼠肝癌細胞Hepa 1-6懸液尾靜脈注射可建立小鼠肝癌CTC模型,可用于抑制循環(huán)腫瘤細胞藥物的篩選與評價。
[Abstract]:Objective to establish a mouse model of circulating tumor cells (CTCs) using mouse hepatoma Hepa 1 / 6 cells. Methods one hundred and eight male C57BL/6 mice were divided into three groups according to body weight, 36 mice in each group. 3 groups were inoculated with 1 脳 10 6, 5 脳 10 6 and 1 脳 10 7 cells / mL of single cell suspension of hepatoma Hepa 1 / mL in each of the three groups by intravenous inoculation of 0.2 mL, 5 脳 10 6 and 1 脳 10 7 cells / mL, respectively. Blood samples were collected 1,5,9,13,17,21 days after inoculation in each group. Flow cytometry was used to calculate the number and proportion of CTCs in 20 000 nucleated cells, the relative CTCs inhibition rate and animal mortality rate were recorded. (2) 80 male C57BL/6 mice were used. According to body weight, they were divided into two groups: model control group and sulafinib group with 40 rats in each group. The single cell suspension of Hepa 1 / 6 cell was injected into each tail vein at a concentration of 5 脳 10 6 cells / mL. On the second day after inoculation, the rats were intragastrically administrated with 50 mg/kg toluene sulfonate for 21 days, and then given 3,8, 8, 8, 10 6 cells per tail vein for 21 consecutive days, with a concentration of 5 脳 10 6 cells / mL single cell suspension. Blood samples were collected for CTC test on day 15 and 21. Results (1) at 1,5,9,13,17,21 days after inoculation with 1 脳 10 6 cells / mL of cell suspension, the percentage of CTCs in animals was 25.1%, 18.1%, 8.9%, 4.4%, 2.9%, 0.3%, and no animal death was observed. At 1,5,9,13,17,21 days after inoculation with 5 脳 10 ~ 6 cells / mL of cell suspension, the percentage of CTCs in animals was 40.4%, 35.4%, 15.4%, 9.0%, 6.6%, 4.1%, and no animal death was observed. After inoculation with 1 脳 10 ~ 7 cells / mL cell suspension, the CTCs ratio of animals on day 1 and 5 was 39.1%, 33.5%, and immediately after inoculation, animal death occurred. All animals died on the 7th day after inoculation. (2) the clearance rates of D3, D8, D15 and D21 were-7.5%, 4.6%, 55.3% and 94.5%, respectively, during the administration of sulafinib toluene sulfonate, and the relative circulating tumor cell clearance rates were-7.5%, 4.6%, 55.3%, 94.5%, respectively. Compared with the model control group, there was significant difference (P0.05 or P0.01). Conclusion the mouse hepatoma CTC model can be established by intravenous injection of 5 脳 10 6 / mL of mouse hepatoma cell line Hepa 1 / mL, which can be used to screen and evaluate the drugs for inhibiting circulating tumor cells.
【作者單位】： 湖南省藥物安全評價研究中心新藥藥效與安全性評價湖南省重點實驗室;中南大學湘雅醫(yī)院血液科;
【基金】：湖湘青年創(chuàng)新創(chuàng)業(yè)平臺專項(2014) 湖南省科技計劃重點項目(2012TT1002)
【分類號】：R-332;R735.7
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本文編號：2448253