小剂量阿司匹林协同干扰素-α抑制肝细胞肝癌生长转移的实验研究

The role and mechanism of low-dose aspirin with IFN-α in inhibiting growth and metastasis of hepatocellular carcinoma

【摘要】目的探讨小剂量阿司匹林协同干扰素-α（IFN-α）抑制肝细胞癌（hepatocellulαr carcinoma，HCC）生长转移的作用及机制。方法培养具有肺转移潜能的人MHCC97L肝癌细胞。将MHCC97L肝癌组织块种植于BALB／c nu／nu雄性裸鼠肝脏，建立人肝癌裸鼠原位模型。以不同剂量组合的阿司匹林和IFN-α作用于荷瘤裸鼠，测量肿瘤体积，计算肺转移灶数目及肺转移率。用MTT及明胶酶谱实验检测阿司匹林对MHCC97L细胞增殖及金属蛋白酶2（matrix metalloprotein ases 2，MMP-2）活性的影响。用Western Blot及ELISA检测细胞及血清MMP-2及血管内皮生长因子（vascular endothelial growth factor，VEGF）蛋白水平。结果对照组肿瘤体积为（3．12±0．85）am。，肺转移率为66．7％。大剂量阿司匹林[45mg／（kg·d）]治疗组肿瘤体积为（1.89±0．88）cm。（P〉0．05），肺转移率为58．3％（P〉0．05）。而大剂量IFN-α[1．5×10^7／（kg·d）]治疗组、大剂量IFN-α＋大剂量阿司匹林治疗组、小剂量IFN-α[7．5×10^6／（kg·d）]＋小剂量阿司匹林[15mg／（kg·d）]治疗组肿瘤体积分别为（0．69±0．40）cm^3、（0．55±0．31）cm^3、（0．40±0．43）cm^3，均显著低于对照组（P〈0．05），肺转移率均为0（P〈0．05）。2mmol／L阿司匹林对MHCC97L细胞增殖无显著影响（P〉0．05），但可抑制其MMP-2的活性及VEGF的水平。小剂量IFN-α＋小剂量阿司匹林治疗组裸鼠血清MMP-2及VEGF显著降低（P〈0．05）。结论小剂量阿司匹林可协同IFN-α抑制HCC生长转移，抑制MMP-2和VEGF的活性和表达是其重要作用机制之一。

Objective To study the role and mechanism of low-dose aspirin with IFN-a in inhibiting growth and metastasis of hepatocellular carcinoma （HCC）. Methods MHCC97L cells were cultured and a metastatic model of human HCC was established by orthotopic implantation of histologically intact human HCC tissue into the liver of nude （nu/nu） mice. After administration of different doses of Aspirin and IFN-a for 40 days, the mice hearing xenografts in liver were killed, and the tumor volume and lung metastasis were evaluated. Cell proliferation and MMP-2 activity were measured by MTT and gelatin zymography, respectively. The expressions of VEGF and MMP-2 were measured by western blot and ELISA. Results Compared to the control group, there were no significant differences in the high-dose Aspirin [45 mg/（kg·d）] treated group regarding tumor volume [（1.89＋ 0.88） cm3 vs （3.12±0.85） cm^3 , P〉0.05] and incidence of lung metastases （58.3% vs 66.7%, P〉 0.05）, but the tumor volume and incidence of lung metastasis were significantly inhibited in the high- dose IFN-a group [1.5 ×10^7/（kg·d）], the high-dose IFN-α combined with high-dose Aspirin group, and the low-dose IFN-α [7.5× 10^6/（kg·d）] combined with low-dose Aspirin [15 mg/（kg·d] group （P〉0.05）. 2 mmol/L Aspirin did not inhibit the proliferation of MHCC97 cells （P〉0.05）, but in- hibited the activities and expressions of MMP-2 and VEGF. Low-dose IFN-a combined with low-dose Aspirin significantly decreased the expressions of MMP-2 and VEGF in nude mice （P%0.05）. Conclusion Low-dose Aspirin combined with low-dose IFN-a significantly inhibited the growth and metastasis of HCC through suppressing the expressions of MMP-2 and VEGF.