CIP-13F抗肿瘤转移作用及机制的探讨  

Inhibition effect of CIP-13F on metastasis and its mecgabusn

作　　者：张厚莉[1,2] 裴可灵[1,2] 崔淑香[1,2] 

机构地区：[1]山东省医学科学院药物研究所 [2]济南大学.山东省医学科学院医学与生命科学学院山东省罕少见病重点实验室,山东济南250062

出　　处：《中华肿瘤防治杂志》2013年第6期405-410,共6页Chinese Journal of Cancer Prevention and Treatment

基　　金：国家自然科学基金(81072665)

摘　　要：目的:研究APN抑制剂CIP-13F抗肿瘤侵袭转移及血管生成的作用机制,为药物开发和恶性肿瘤临床治疗奠定实验基础。方法:体外培养人卵巢透明细胞癌ES-2和人纤维肉瘤细胞HT-1080,应用L-亮氨酰对硝基苯胺底物法评价CIP-13F对APN酶活性的抑制作用,通过MTT比色评价CIP-13F对肿瘤细胞的生长抑制作用。以Transwell cham-ber法观察CIP-13F对ES-2细胞游走能力的抑制作用。体内建立Lewis肺癌自发性肺转移模型,分别给药后取肿瘤组织和肺组织,应用免疫组织化学法检测Lewis肺癌组织中APN的表达水平。以免疫组织化学法检测CD34表达水平评价肿瘤微血管密度(MVD)。结果:采用4、20、100和500μmol/L CIP-13F作用于ES-2和HT-1080细胞发现,对ES-2细胞,APN的表达和生长抑制作用呈现剂量-效应相关性,对HT-1080细胞中APN和生长抑制作用较弱。另外,不同浓度的CIP-13F与ES-2细胞穿过人工基底膜的能力呈剂量依赖性关系。100和500μmol/L的CIP-13F对肿瘤细胞穿过Matrigel的游走能力抑制率分别为42.0%和72.1%,P值分别为0.003和0.001。体内试验表明,Bestatin 100mg/kg组、CIP-13F50和100mg/kg剂量的肿瘤生长抑制率分别为33.2%、29.4%和45.8%(P值分别为0.013、0.020和0.005),同时,与阴性对照组相比,50和100mg/kg CIP-13F对Lewis肺癌肺转移抑制率分别为52.2%和81.3%(P值分别为0.001和0.000)。另外,50和100mg/kg CIP-13F作用下,Lewis肺癌组织的MVD值分别为18.6和14.9,与阴性对照组比较,MVD值均明显降低,P值分别为0.000和0.001。结论:环酰亚胺类肽化合物CIP-13F通过抑制APN活性,抑制肿瘤细胞的增殖和转移,进而抑制移植瘤鼠的腋部原发肿瘤灶的生长,降低肺继发转移灶的数目。CIP-13F用于治疗APN表达阳性的肿瘤有良好的应用前景。OBJECTIVE： To evaluated the efficacy of CIP-13F as a novel APN inhibitor for treatment of cancers with positive APN expression. METHODS： Ovarian clear cells carcinoma ES-2 and fibrosareoma cells HT-1080 in vitro,the as- say of quantitating the enzymatic cleavage of the substrate L-Leucine p-nitroanilide was employed to evaluate APN activi- ty. MTT assay was employed to evaluate the efficacy of CIP-13F in vitro. The transwell chamber assay was used to meas- ure the inhibitory effect of CIP-13F on the invasion and migration of cancer cells penetrating the Matrigel. The in vivo ef- ficacy of CIP-13F was assessed in an Lewis lung carcinoma （LLC） implantation mouse model,and the assays of immuno- histochemical staining and western blotting were performed to estimate the expression of APN in LLC cells. Using immu- nohistochemical staining with CD34,the antiangiogenesis of CIPq3F was evaluated in LLC tissue sections. RESULTS： In vitro showed that CIP-13F inhibited ES-2 growth and migration via suppression of APN, whereas, CIP-13F had weak in- hibition on the growth and the expression of APN of HT-1080. The ability through artificial basement membrane of dif- ferent concentrations of CIP-13F exhibited a dose-dependent manner,the inhibition rates of 100 and 500 μmol/L CIP-13Fwere 42.0 % and 72.1% （P= 0. 003, P= 0. 001）. Further more to evaluated the inhibitory effect of CIP-13F in mice bear ing Lewis lung carcinoma implantation was evaluated too. Bestatin 100 mg/kg,CIP-13F 50 mg/kg and 100 mg/kg treat- ment resulted in a significant delay of LLC growth in anterior flank and the tumor inhibition rates were 33.2 %, 29.4 and 45.8% （P values were 0. 013,0. 020 and 0. 005）. Compared with the control,the inhition rates of Lewis lung metas- tasis of CIP-13F 50 mg/kg and 100 mg/kg treatment were 52.2% and 81.3% （P values were 0. 001 and 0. 000）. Further more the inhibition of angiogenesis in LLC tissues was determined, after treatment of 50 mg/kg and 100 mg/kg CIP-13F, the MVD values of Lewis lung tissue we

关 键 词：APN CD13 CIP-13F 肿瘤侵润 肿瘤转移 血管生成 

分 类 号：R96[医药卫生—药理学]