安罗替尼联合5-氟尿嘧啶抑制结肠癌的增殖和血管生成

Anlotinib combined with 5-fluorouracil inhibits proliferation and angiogenesis of colon cancer

目的探讨安罗替尼联合5-氟尿嘧啶对结肠癌CT26细胞的增殖、侵袭、凋亡及血管生成的影响。方法在体外实验,应用MTT法、Transwell法以及流式细胞术检测安罗替尼组(4μmol/L)、5-氟尿嘧啶组(5μg/mL)以及联合组(安罗替尼4μmol/L+5-氟尿嘧啶5μg/mL)对结肠癌CT26细胞的增殖、侵袭及凋亡的影响。然后,建立结肠癌CT26细胞的皮下移植瘤模型,随机分为4组(n=12):NS组,安罗替尼组(2 mg/kg),5-氟尿嘧啶组(20 mg/kg),联合组(安罗替尼2 mg/kg+5-氟尿嘧啶20 mg/kg),探究各组对结肠癌皮下移植瘤的抗肿瘤作用,观察各组带瘤小鼠的肿瘤生长情况,采用免疫组化检测各组肿瘤组织中Ki-67的表达水平和微血管密度。结果体外细胞实验结果显示联合组对细胞增殖活性的抑制率明显高于单药安罗替尼组或5-FU组(P<0.05),且联合组对结肠癌CT26细胞侵袭能力的抑制作用最大(P<0.05);此外,流式细胞术检测发现联合组使细胞凋亡率显著增高(P<0.05)。皮下移植瘤模型实验结果显示联合组能明显抑制小鼠皮下移植瘤的生长(P<0.05)。与生理盐水组、单药安罗替尼或5-FU组相比,联合组药物作用能显著降低肿瘤组织中的Ki-67阳性表达率和微血管密度(P<0.05)。结论安罗替尼联合5-氟尿嘧啶能显著抑制结肠癌的增殖和侵袭,促进细胞的凋亡,并抑制小鼠肿瘤组织中细胞的增殖及血管的生成,具有良好的抗肿瘤作用。

Objective To investigate the effects of Anlotinib combined with 5-fluorouracil on the proliferation,invasion,apoptosis and angiogenesis of colon cancer CT26 cell line.Methods In vitro,MTT assay,Transwell assay and flow cytometry were used to detect the effects of the Anlotinib(4μmol/L),5-fluorouracil(5μg/ml)and the combined drugs(Anlotinib 4μmol/L+5-fluorouracil 5μg/ml)on the proliferation,invasion and apoptosis of CT26 cells.The subcutaneous transplanted tumor model of colon cancer was established and randomly divided into four groups(n=12):Normal saline(NS)group,Anlotinib(2 mg/kg)group,5-fluorouracil(20 mg/kg)group and the combined group(Anlotinib 2 mg/kg+5-fluorouracil 20 mg/kg).The tumor growth of tumor bearing mice in each group was observed.The expression of Ki-67 and microvessel density was detected by immunohistochemistry.Results The inhibition rate of cell activity in the combined group was significantly higher than that in Anlotinib group or the 5-FU group.The combined group had the greatest inhibition effects on the invasion ability of colon cancer CT26 cells.In addition,the apoptosis rate of the combined group was significantly higher than that in any drug alone group in vitro.The combined group significantly inhibited the growth of subcutaneous transplanted tumor in mice.Compared with the NS group,the Anlotinib or the 5-FU group,the combined group significantly reduced the positive expression rate of Ki-67 and microvessel density in tumor tissues.Conclusion Anlotinib combined with 5-fluorouracil could significantly inhibit the proliferation and invasion of colon cancer,promote the apoptosis of cells,suppress the angiogenesis in tumor tissues of mice.