目的 探讨基质金属蛋白酶-1(MMP-1)和膜型1基质金属蛋白酶(MT1-MMP)在胆囊癌血管生成拟态中的表达及临床意义。方法 收集手术病理确诊的原发性胆囊癌74例和胆囊腺瘤、慢性胆囊炎各10例的石蜡标本及相关临床资料,应用免疫组化测定上述病变组织的MMP-1、MT1-MMP表达。应用Kaplan-Meier生存比较、Cox风险模型分析与胆囊癌血管生成拟态(VM)的相关性和临床意义。结果 ① 胆囊癌MMP-1、MT1-MMP表达明显高于胆囊腺瘤、胆囊炎(P<0.000 1);VM(+)胆囊癌MT1-MMP表达明显高于VM(-)胆囊癌(P=0.003 9),而MMP-1则无差别。② MMP-1表达与VM(-)胆囊癌的Nevin 分期(P=0.003 6)、分化程度(P=0.010)、肝转移(P=0.003)和淋巴结转移(P=0.002)正相关;MT1-MMP表达与VM(-)或VM(+)胆囊癌的Nevin 分期(P=0.013或P=0.033)、浸润深度(P=0.045或P=0.035)、淋巴结转移(P=0.046或P=0.025)和肝转移(P=0.030或P=0.027)正相关;MMP-1、MT1-MMP表达与VM(-)胆囊癌正相关。③ VM(+)胆囊癌MT1-MMP表达在Nevin S3~S5期(P=0.000 1)、侵犯浆膜(P=0.001)、淋巴结转移(P=0.000 2)和肝转移(P=0.004)同样分组条件下分别明显高于VM(-)胆囊癌;MMP-1仅在肝转移组,VM(+)胆囊癌表达显著高于VM(-)(P=0.038)。④ 无论在MMP-1、MT1-MMP单独表达阳性或二者均阳性,VM(+)胆囊癌患者术后5年生存期都明显短于VM(-)组(P=0.025 1或P=0.022 5或P=0.025 0)。Cox多因素分析表明,VM、浸润深度、淋巴结转移、肝转移、手术方式是影响胆囊癌患者预后的独立因素。结论 胆囊癌MMP-1、MT1-MMP高表达,MT1-MMP表达与胆囊癌VM相关,MMP-1则与胆囊癌VM无关。MMP-1、MT1-MMP可作为评判胆囊癌Nevin 分期、浸润深度、淋巴结或肝转移及判断预后的重要指标;而MT1-MMP还可作为判断胆囊癌否存在VM和VM胆囊癌预后的重要指标。
Objective To evaluate the expression of matrix metalloproteinase-1(MMP-1) and membrane type-1 matrix metalloproteinase (MT1-MMP) in human primary gallbladder carcinomas with vasculogenic mimicry (VM) and its clinical significance.Methods Seventy-four carcinomas,10 adenomas and 10 chronic inflammatory lesions of the gallbladder underwent operation and confirmed histopathologically and their clinical-pathlogical data were studied and followedup.Immunohistochemistry (SABC) was used to determine the expression of MMP-1,MT1-MMP proteins in each paraffin section of every patient in vivo.Kaplan-Meier survival curves and Cox multiple factor were used to analyse the relation beteewn VM of gallbladder carcinomas and expression of MMP-1,MT1-MMP.Results ① Expression of MMP-1 and MT1-MMP was increased significantly in carcinomas with or without VM than adenomas and inflammatory lesions of the gallbladder (P<0.000 1) in.Eexpression of MT1-MMP in the gallbladder carcinomas with VM was increased significantly than that without VM (P=0.003 9),without difference on expression of MMP-1 between VM (+) gallbladder carcinomas and VM (-) gallbladder carcinomas.② There is positive correlation between expression of MMP-1 and VM (-) gallbladder carcinomas in the cases of Nevin stage (P=0.003 6),differentiation degree (P=0.010),liver metastatis (P=0.003) and lymph node metastases (P=0.002),the same correlation was observed between expression of MT1-MMP and VM (+)/(-) gallbladder carcinomas in the cases of Nevin stage (P=0.013 or P=0.003 3),invasion depth (P=0.045 or P=0.035),lymph node metastases (P=0.046 or P=0.025) and liver metastatis (P=0.030 or P=0.027); MMP-1 expression was positively correlated with MT1-MMP expression in VM (-) gallbladder carcinomas.③ Expression of MT1-MMP in the same conditions of Nevin stage (S3~S5,P=0.000 1),serosal invasion (P=0.001),lymph node metastases (P=0.000 2),liver metastatis (P=0.004) was higher significantly in VM (+) gallbladder carcinomas than VM (-) gallbladder carcinomas; Furthermore,expression of MMP-1 in the cases of liver metastatis (P=0.0038) was higher significantly in VM (+) gallbladder carcinomas than VM (-) gallbladder carcinomas.④ VM (+) gallbladder carcinoma group underwent operation with positive expression of MMP-1 and MT1-MMP had shorter 5-year survival than the respectively VM (-) group (P=0.025 1; P=0.022 5),the same correlation was observed in positive coexpression of MMP-1 and MT1-MMP (P=0.025 0).Cox analysis showed that VM,invasion depth,lymph node metastasis,hepatic metastasis,and operation method were independent risk factors of the prognosis of patients with primary gallbladder carcinoma.Conclusions Higher expression of MMP-1,MT1-MMP in gallbladder carcinomas,a positive correlation between expression of MT1-MMP and VM of gallbladder carcinomas and no correlation between expression of MMP-1 and VM of gallbladder carcinomas were observed.MMP-1 and MT1-MMP are important clinical markers for staging of Nevin stage,judgement of invasion depth,lymph node or liver metastatis,and evaluation of prognosis in patients with gallbladder carcinoma.MT1-MMP may be especially of important for evaluating if there is VM in human gallbladder carcinomas and prognosis of patients with VM (+) gallbladder carcinoma.
[1] Maniotis AJ,Folberg R,Hess A,et al.Vascular channel formation by human melanoma cells in vivo and in vitro:vasculogenic mimicry[J].Am J Pathol,1999,155(3):739-752.
[2] Shirakawa K,Wakasugi H,Heike Y,et al.Vasculogenic mimicry and pseudo-comedo formation in breast cancer[J].Int J Cancer,2002,99(6):821-828.
[3] Cai Xuansong,Jia Yongwei,Mei Jiong,et al.Tumor blood vessels formation in osteosarcoma:vasculogenesis mimicry[J].Chin J Med,2004,117(1):94-98.
[4] 赵秀兰,杜静,张诗武,等.肝细胞癌中血管生成拟态的研究[J].中华肝脏病杂志,2006,14(1):41-44.
[5] 范跃祖,孙伟,张文忠,等.原发性胆囊癌患者肿瘤血管生成拟态及其临床意义[J].中华医学杂志,2007,87(3):145-149.
[6] Herbst RS,Yano S,Kuniyasu H,et al .Differential expression of E-cadherin and type IV collagenase genes predicts outcome in patients with stage I non-small cell lung carcinoma[J].Clin Cancer Res,2000,6(3):790-797.
[7] Hess AR,Postovit LM,Margaryan NV,et al.Focal adhesion kinase promotes the aggressive melanoma phenotype[J].Cancer Res,2005,65(21):9851-9860.
[8] Hendrix MJ,Seftor EA,Meltzer PS,et al.Expression and functional significance of VE-cadherin in aggressive human melanoma cells:role in vasculogenic mimicry[J].Proc Natl Acad Sci,2001,98(14):8018-8023.
[9] Hess AR,Seftor EA,Gardner LM,et al.Molecular regulation of tumor cell vasculogenic mimicry by tyrosine phosphorylation:role of epithelial cell kinase (Eck/EphA2)[J].Cancer Res,2001,61(8):3250-3255.
[10] Hess AR,Seftor EA,Seftor RE,et al.Phosphoinositide 3-kinase regulates membrane Type 1-matrix metalloproteinase (MMP) and MMP-2 activity during melanoma cell vasculogenic mimicry[J].Cancer Res,2003,63(16):4757-4762.
[11] Sood AK,Fletcher MS,Coffin JE,et al.Functional role of matrix metalloproteinases in ovarian tumor cell plasticity[J].Am J Obstet Gynecol,2004,190(4):899-909.
[12] Seftor RE,Seftor EA,Kirschmann DA,et al.Targeting the tumor microenvironment with chemically modified tetracyclines:inhibition of laminin 5 gamma 2 chain promigratory fragments and vasculogenic mimicry[J].Mol Cancer Ther,2002,1(13):1173-1179.
[13] Seftor RE,Seftor EA,Koshikawa N,et al.Cooperative interactions of laminin 5 gamma 2 chain,matrix metalloproteinase-2,and membrane type-1-matrix/metalloproteinase are required for mimicry of embryonic vasculogenesis by aggressive melanoma[J].Cancer Res,2001,61(17):6322-6327.
[14] Ruf W,Seftor EA,Petrovan RJ,et al.Differential role of tissue factor pathway inhibitors 1 and 2 in melanoma vasculogenic mimicry[J].Cancer Res,2003,63(17):5381-5389.
