血管生成抑制剂YH-16联合氟尿嘧啶抑制结直肠癌肝转移的研究

Inhibitory effect of angiogenesis inhibitor YH-16 in combination with 5-FU on liver metastasis of colorectal cancer

目的探讨血管生成抑制剂YH-16和氟尿嘧啶(5-FU)联合应用对结直肠癌肝转移的抑制作用。方法用MTT方法测定血管生成抑制剂YH-16和5-FU对血管内皮细胞和结肠癌细胞的IC50;建立小鼠肝转移模型,随机分为对照组、YH-16组(又分为低、中、高剂量3组)和5-FU组及联合治疗组(YH-16加5-FU),术后2周观察各组小鼠肝转移瘤数目、原发灶大小和毒性反应,并检测肝转移瘤血管内皮生长因子(VEGF)的表达和肿瘤微血管密度(MVD)。结果YH-16对结肠癌细胞的IC50是血管内皮细胞的3.38倍,而5-FU对两种细胞的IC50差别不大。高剂量YH-16组、5-FU组和联合治疗组肝转移瘤数目明显低于对照组,而联合治疗组又低于高剂量YH-16组和5-FU组(均P<0.05)。YH-16各剂量组的脾原发瘤体积与对照组比较均P>0.05,差异无统计学意义;而5-FU组和联合治疗组则小于对照组(均P<0.05)。YH-16的毒性明显低于5-FU(P<0.05),且两者联合使用其毒性与单用5-FU比较,差异无统计学意义(P>0.05)。5-FU组和联合治疗组肝转移瘤组织中VEGF的表达明显降低,中、高剂量YH-16组和5-FU组及联合治疗组肝转移瘤组织中的MVD计数明显降低(均P<0.05)。结论血管生成抑制剂YH-16明显抑制结直肠癌肝转移,YH-16与5-FU联合应用对结直肠癌肝转移的抑制具有协同作用。

Objective To study the effect of angiogenesis inhibitor YH-16 in combination with 5-FU on liver metastasis of colorectal cancer. Methods In vitro, the inhibitory effects of YH-16 and 5-FU on the growth of vascular endothelial cells and colorectal cancer cells were examined by MTT assay. In vivo, cohrectal cancer cells were transplanted into BALB/c mice, and the mice were divided into six groups randomly： control group, low-dose YH-16 group, middle-dose YH-16 group, high-dose YH-16 group, 5-FU group and combination group. The number of liver metastases, the size of primary tumor and the toxicity were examined after 2 weeks postoperatively. The expression of vascular endothelial growth factor （VEGF） in liver metastases was detected by immunohistochemistry, and tumor microvessel density （MVD） was measured by immunostaining with CD34 and factor Ⅷ monocolonal antibodies. Results In vitro, YH-16 inhibited the growth of colon cancer cells and vascular endothelial cells, with the IC50 at （2. 16 ±0.28） μg/ml and （0.64 ±0. 10）μg /ml respectively. In vivo high-dose YH-16 and 5-FU had a remarkable inhibitory effect on liver metastasis, and the combination group showed significant enhancement on this effect （ P 〈 0. 05）. The combination group and 5-FU group could inhibit the growth of primary tumor, but not found in YH-16 group. The toxicity of YH-16 was lower than that of 5-FU （ P 〈 0. 05）, and the difference was not found in the toxicity between combination group and 5-FU group （P 〉 0.05） . Expression of VEGF in liver metastases was clearly inhibited by YH-16 in combination with 5-FU or 5-FU alone compared to the control group, and MVD in middle-dose and high-dose YH-16 group, 5-FU group and combination group was lower than that in control group （ P 〈 0. 05）. Conclusions The angiogenesis inhibitor YH-16 can inhibit liver metastasis of colorectal cancer through inhibiting the growth of vascular endothelial cells. YH-16 in combination with 5-FU has additive effect on inhibitory activity against liver metastasis.