CAJ | 학술논문

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(hepatocellular 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 metalloproteinases 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)cm3,肺转移率为66.7％.大剂量阿司匹林[45 mg/(kg· d)]治疗组肿瘤体积为(1.89±0.88)cm3 (P＞0.05),肺转移率为58.3％ (P＞0.05).而大剂量IFN-α[1.5×107/(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 [7.5×106/(kg·d)]+小剂量阿司匹林15 mg/(kg· d)]治疗组肿瘤体积分别为(0.69±0.40)cm3、(0.55±0.31)cm3、(0.40±0.43)cm3,均显著低于对照组(P＜0.05),肺转移率均为0(P＜0.05).2 mmol/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.
목적 탐토소제량아사필림협동간우소-α(IFN-α)억제간세포암(hepatocellular carcinoma,HCC)생장전이적작용급궤제.방법 배양구유폐전이잠능적인MHCC97L간암세포.장MHCC97L간암조직괴충식우BALB/c nu/nu웅성라서간장,건립인간암라서원위모형.이불동제량조합적아사필림화IFN-α작용우하류라서,측량종류체적,계산폐전이조수목급폐전이솔.용MTT급명효매보실험검측아사필림대MHCC97L세포증식급금속단백매2(matrix metalloproteinases 2,MMP-2)활성적영향.용Western Blot급ELISA검측세포급혈청MMP-2급혈관내피생장인자(vascular endothelial growth factor,VEGF)단백수평.결과 대조조종류체적위(3.12±0.85)cm3,폐전이솔위66.7％.대제량아사필림[45 mg/(kg· d)]치료조종류체적위(1.89±0.88)cm3 (P＞0.05),폐전이솔위58.3％ (P＞0.05).이대제량IFN-α[1.5×107/(kg·d)]치료조、대제량IFN-α+대제량아사필림치료조、소제량IFN-α [7.5×106/(kg·d)]+소제량아사필림15 mg/(kg· d)]치료조종류체적분별위(0.69±0.40)cm3、(0.55±0.31)cm3、(0.40±0.43)cm3,균현저저우대조조(P＜0.05),폐전이솔균위0(P＜0.05).2 mmol/L아사필림대MHCC97L세포증식무현저영향(P＞0.05),단가억제기MMP-2적활성급VEGF적수평.소제량IFN-α+소제량아사필림치료조라서혈청MMP-2급VEGF현저강저(P＜0.05).결론 소제량아사필림가협동IFN-α억제HCC생장전이,억제MMP-2화VEGF적활성화표체시기중요작용궤제지일.
Objective To study the role and mechanism of low-dose aspirin with IFN-α 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-α for 40 days,the mice bearing 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) cm3,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 highdose IFN-α group [1.5 × 107/(kg · d)],the high-dose IFN-α combined with high-dose Aspirin group,and the low-dose IFN-α [7.5 × 106 / (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 inhibited the activities and expressions of MMP-2 and VEGF.Low-dose IFN-α 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-α significantly inhibited the growth and metastasis of HCC through suppressing the expressions of MMP-2 and VEGF.