血管生成抑制剂YH-16抑制结肠癌肝转移的研究

Inhibitory Effect of Angiogenesis Inhibitor YH-16 on Liver Metastases from Colorectal Cancer

背景与目的:YH-16是新合成的重组人血管内皮抑制素,Ⅱ期临床试验已证实YH-16联合化疗治疗晚期非小细胞肺癌具有协同作用。本文探讨血管生成抑制剂YH-16对结肠癌肝转移的抑制作用。方法:MTT法测定血管生成抑制剂YH-16对血管内皮细胞和结肠癌CT26细胞的IC50;经小鼠脾脏下极包膜下注入CT26细胞建立结肠癌肝转移模型,60只小鼠随机分为对照组、低剂量YH-16组、中剂量YH-16组、高剂量YH-16组,YH-16剂量分别为0mg/kg、0.40mg/kg、0.75mg/kg和1.50mg/kg,术后2周观察各组小鼠肝转移情况,采用免疫组化方法检测肝转移瘤组织中血管内皮生长因子(vascularendothelialgrowthfactor,VEGF)的表达和肿瘤微血管密度(microvesseldensity,MVD)。结果:(1)YH-16对结肠癌CT26细胞和血管内皮细胞的IC50分别为(2.16±0.28)μg/ml和(0.64±0.10)μg/ml,前者是后者的3.38倍;(2)对照组,低、中、高剂量YH-16组肝转移率分别为100.0%、92.3%、80.0%和73.3%。高剂量YH-16组肝转移瘤数目明显低于对照组、低剂量YH-16组和中剂量YH-16组(P值均<0.05);(3)对照组,低、中、高剂量YH-16组脾脏肿瘤体积的中位数分别为1.180cm3、1.201cm3、0.887cm3和0.781cm3,四组比较无显著性差异(P>0.05);(4)YH-16各剂量组肝转移瘤组织中VEGF的表达较对照组无明显降低,四组肝转移瘤的MVD分别为65.00±9.58、58.15±8.81、51.60±7.10和44.53±11.47,中、高剂量YH-16组的MVD计数较对照组明显降低,高剂量YH-16组MVD计数较中剂量和低剂量YH-16组明显降低(P值均<0.05)。结论:血管生成抑制剂YH-16可以明显抑制结肠癌肝转移。

BACKGROUND ＆ OBJECTIVE, YH-16, a new recombinant angiogenesis inhibitor, has demonstrated synergetic effects with chemotherapy in non-small-cell lung cancer treatment in stage II clinical trial. This study was to investigate the effect of YH-16 on liver metastases from colon cancer. METHODS： Inhibitory concentration 50% （IC50） of YH-16 on vascular endothelial cells and colon cancer cell line CT26 was determined. by MTT assay. Furthermore, the mouse mode of colon cancer liver metastases was established by inoculating CT26 cells into the subcapsule of spleen. 60 mice were randomly divided into four groups： control group （0 mg/kg）, low-dose YH-16 group （0.40 mg/kg）, medium-dose YH-16 group （0.75 mg/kg） and high-dose YH-16 group （1.5 mg/kg）. The numbers of liver metastases were examined 2 weeks after drug injection. The expression of vascular endothelial growth factor （VEGF） was detected by immunohistochemical method in liver metastases, and tumor microvessel density （MVD） was measured by immunostaining using factor Vii monocolonal antibody. RESULTS. Proliferation of CT26 and vascular endothelial cells was inhibited by YH-16, which the IC5o was （2.16±0.28） μg/ml and （0.64±0.10） g /ml, respectively. In vivo, the liver metastasis rates in control, low-dose, medium-dose and high-dose groups were 100%, 92.3%,80% and 73.3%, respectively （P〈0.05）. However, YH-16 did not inhibit the growth of spleen tumors of which median volumes were 1.180 cm^3, 1.201 cm^3, 0.887 cm^3 and 0.781 cm^3, respectively （P〉0.05）. There was no difference of VEGF expression in liver metastases among the four groups. Moreover, MVD was 65.00±9.58, 58.15±8.81, 51.60±7.10 and 44.53±11.47 in the four groups. MVD in medium-dose and high-dose YH-16 groups was lower than that in control group and MVD was lower in high-dose group than that in medium-dose and low-dose groups （P 〈0.05）. CONCLUSION. Angiogenesis inhibitor YH-16 can inhibit liver metastases from colorectal cancer.