胆囊癌血管生成拟态,缺氧诱导因子1α和血管内皮生长因子表达及其与临床预后的关系

摘要/Abstract

摘要： 目的探讨人胆囊癌是否存在血管生成拟态(VM)以及VM、缺氧诱导因子-1α(HIF-1)和血管内皮生长因子(VEGF)与胆囊癌预后的关系。方法应用HE和CD31-PAS 染色观察胆囊癌VM, 应用免疫组织化学、计算机图像、接收机操作特性曲线和高低免疫组化反应截止评分定量分析胆囊癌HIF-1α 、VEGF表达;同时评估HIF-1α、VEGF表达与胆囊癌VM 和无VM组病人临床病理学特征与预后的关系。结果 21.35%的胆囊癌存在VM, VM与胆囊腺癌和肝转移相关(P<0.01)。VM阳性胆囊癌病人存在HIF-1α高表达和VEGF低表达(P<0.01)。HIF-1α 与胆囊癌VM病人的浆膜浸润、分化程度(P<0.01)和VM亚组病人的Nevin分期、阴性肝转移和浆膜浸润、阴性和阳性淋巴结转移(P<0.05)相关。VEGF则与胆囊癌VM病人的肝转移、分化程度(P<0.01)和VM亚组病人Nevin分期、阴性肝转移和中分化程度(P<0.01)相关。多因素分析证实, VM、Nevin 分期和HIF-1α 是胆囊癌病人的独立因素。结论人胆囊癌存在VM; HIF-1α 和VEGF表达与胆囊癌病人的VM和疾病进展明显相关;VM 和IF-1α是胆囊癌病人预后判断的独立因素。

关键词: 血管生成拟态, 缺氧诱导因子-1α, 血管内皮生长因子, 胆囊肿瘤, 预后

Abstract: Purpose To explore whether VM exists in human gallbladder carcinoma (GBC) and VM’ correlation with HIF-1α,VEGF and prognosis of GBC patients.Methods HE and CD31-PAS staining were used to observe VM in GBC.Immunohistochemistry (IHC),computer-assisted image and receiver operating characteristic curve were used to analyze expression of HIF-1α and VEGF in GBC,IHC quantitative analysis and selection of the cut-off score for high or low IHC reactivity.Correlation of HIF-1α and VEGF expression with clinicopathologic features and prognosis of GBC patients with VM and non-VM was evaluated simultaneously.Results VM was observed in GBC (21.35%) and associated with adenocarcinoma and liver metastasis (both P<0.01).Over-expression of HIF-1α and reduced-expression of VEGF were observed in GBC patients with VM (both P<0.01).HIF-1α was correlated with serosal invasion,degree of differentiation in VM group (all P<0.01),and VM in the same subset of clinical-pathologic parameters including Nevin stage,negative liver metastasis and serosal invasion,negative and positive lymph node metastasis (all P<0.05).VEGF was associated with liver metastasis,degree of differentiation in VM group (all P<0.01),and VM in the same subset of clinicopathologic parameters including Nevin stage,negative liver metastasis and moderate degree of differentiation (all P<0.01).Multivariate analysis confirmed that VM,Nevin stage and HIF-1α were independent factors for GBC patients.Conclusions VM exists in human GBC.HIF-1α and VEGF expression is significantly associated with VM and advanced stage in GBC patients.VM and HIF-1α is independent factors for GBC patients.

Key words: Vasculogenic mimicry, Hypoxia inducible factor-1α, Vascular endothelial growth factor, Gallbladder neoplasm, Prognosis

中图分类号:

R575.6

孙伟, 范跃祖, 张文忠, 葛春艳. 胆囊癌血管生成拟态,缺氧诱导因子1α和血管内皮生长因子表达及其与临床预后的关系[J]. 外科研究与新技术, 2012, 1(1): 27-41.

Sun Wei, Fan Yue-Zu, Zhang Wen-Zhong, Ge Chun-Yan. Vasculogenic mimicry,HIF-1α and VEGF expression in gallbladder carcinoma:association with clinical outcomes and prognostic implications[J]. 外科研究与新技术, 2012, 1(1): 27-41.

参考文献

[1] Miller G,Jarnagin WR.Gallbladder carcinoma[J].EJSO,2008,34(3):306-312.
[2] Hsing AW,Gao YT,Devesa SS,et al.Rising incidence of biliary tract cancers in Shanghai,China[J].Int J Cancer,1998,75(3):368-370.
[3] Zhu AX,Hong TS,Hezel AF,et al.Current Management of Gallbladder Carcinoma[J].Oncologist,2010,15(2):168-181.
[4] Kuroki T,Tajima Y,Matsuo K,et al.Genetic alterations in gallbladder carcinoma[J].Surgery Today,2005,35(2):101-105.
[5] 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.
[6] Folberg R,Hendrix MJ,Maniotis AJ.Vasculogenic mimicry and tumor angiogenesis[J].Am J Pathol,2000,156(2):361-381.
[7] Frenkel S,Barzel I,Levy J,et al.Demonstrating circulation in vasculogenic mimicry patterns of uveal melanoma by confocal indocyanine green angiography[J].Eye (Lond),2008,22(7):948-952.
[8] Folberg R,Maniotis AJ.Vasculogenic mimicry[J].APMIS,2004,112(7-8):508-525.
[9] Hendrix MJ,Seftor EA,Hess AR,et al.Vasculogenic mimicry and tumour-cell plasticity:lessons from melanoma[J].Nat Rev Cancer,2003,3(6):411-421.
[10] Lissitzky JC,Parriaux D,Ristorcelli E,et al.Cyclic AMP signaling as a mediator of vasculogenic mimicry in aggressive human melanoma cells in vitro[J].Cancer Res,2009,69(3):802-809.
[11] Vartanian AA,Stepanova EV,Gutorov SL,et al.Prognostic significance of periodic acid-Schiff-positive patterns in clear cell renal cell carcinoma[J].Can J Urol,2009,16(4):4726-4732.
[12] 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.
[13] Basu GD,Liang WS,Stephan DA,et al.A novel role for cyclooxygenase-2 in regulating vascular channel formation by human breast cancer cells[J].Breast Cancer Res,2006,8(6):R69.
[14] Robertson FM,Simeone AM,Lucci A,et al.Differential regulation of the aggressive phenotype of inflammatory breast cancer cells by prostanoid receptors EP3 and EP4[J].Cancer,2010,116(Suppl 11):2806-2814.
[15] Sood AK,Fletcher MS,Zahn CM,et al.The clinical significance of tumor cell-lined vasculature in ovarian carcinoma:implications for anti-vasculogenic therapy[J].Cancer Biol Ther,2002,1(6):661-664.
[16] 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.
[17] Sun B,Zhang S,Zhang D,et al.Vasculogenic mimicry is associated with high tumor grade,invasion and metastasis,and short survival in patients with hepatocellular carcinoma[J].Oncol Rep,2006,16(4):693-698.
[18] Sun BC,Zhang SW,Zhao XL,et l.Vasculogenic mimicry is associated with shorter survival in hepatocellular carcinomas[J].Laboratory Investigation,2006,86:1302.
[19] Liu WB,Xu GL,Jia WD,et al.Prognostic significance and mechanisms of patterned matrix vasculogenic mimicry in hepatocellular carcinoma[J].Med Oncol,2011,28(Suppl 1):S228-238.
[20] Wang W,Lin P,Han C,et al.Vasculogenic mimicry contributes to lymph node metastasis of laryngeal squamous cell carcinoma[J].J Exp Clin Cancer Res,2010,29:60.
[21] El Hallani S,Boisselier B,Peglion F,et al.A new alternative mechanism in glioblastoma vascularization:tubular vasculogenic mimicry.Brain,2010,133(Pt 4):973-982.
[22] Li M,Gu Y,Zhang Z,et al.Vasculogenic mimicry:a new prognostic sign of gastric adenocarcinoma[J].Pathol Oncol Res,2010,16(2):259-266.
[23] Baeten CI,Hillen F,Pauwels P,et al.Prognostic role of vasculogenic mimicry in colorectal cancer[J].Dis Colon Rectum 2009,52(12):2028-2035.
[24] Zhao H,Gu XM.Study on vasculogenic mimicry in malignant esophageal stromal tumors[J].World J Gastroenterol,2008,14(15):2430-2433.
[25] Sun B,Qie S,Zhang S,et al.Role and mechanism of vasculogenic mimicry in gastrointestinal stromal tumors[J].Hum Pathol,2008,39(3):444-451.
[26] Talks KL,Turley H,Gatter KC,et al.The expression and distribution of the hypoxia-inducible factors HIF-1 alpha and HIF-2 alpha in normal human tissues,cancers,and tumor-associated macrophages[J].Am J Pathol,2000,157(2):411-421.
[27] Minet E,Michel G,Remacle J,et al.Role of HIF-1 as a transcription factor involved in embryonic development,cancer progression and apoptosis[J].Int J Mol Med,2000,5(3):253-259.
[28] Seo Y,Baba H,Fukuda T,et al.High expression of vascular endothelial growth factor is associated with liver metastasis and a poor prognosis for patients with ductal pancreatic adenocarcinoma[J].Cancer,2000,88(10):2239-2245.
[29] Huang LE,Arany Z,Livingston DM,et al.Activation of hypoxia-inducible transcription factor depends primarily upon redox-sensitive stabilization of its alpha subunit[J].J Biol Chem,1996,271(50):32253-32259.
[30] Sutter CH,Laughner E,Semenza GL.Hypoxia-inducible factor 1 alpha protein expression is controlled by oxygen-regulated ubiquitination that is disrupted by deletions and missense mutations[J].Proceedings Nat Acad Sci USA,2000,97(9):4748-4753.
[31] Bissell MJ.Tumor plasticity allows vasculogenic mimicry,a novel form of angiogenic switch.A rose by any other name?[J] Am J Pathol,1999,155(3):675-679.
[32] Seftor EA,Meltzer PS,Kirschmann DA,et al.Molecular determinants of human uveal melanoma invasion and metastasis[J].Clin Exp Metastasis,2002,19(3):233-246.
[33] van der Schaft DW,Hillen F,Pauwels P,et al.Tumor cell plasticity in Ewing sarcoma,an alternative circulatory system stimulated by hypoxia[J].Cancer Res,2005,65(24):11520- 11528.
[34] Sun B,Zhang D,Zhang S,et al.Hypoxia influences vasculogenic mimicry channel formation and tumor invasion-related protein expression in melanoma[J].Cancer Lett,2007,249(2):188-197.
[35] Vartanian AA,Burova OS,Stepanova EV,et al.Melanoma vasculogenic mimicry is strongly related to reactive oxygen species level[J].Melanoma Res,2007,17(6):370-379.
[36] Su M,Feng YJ,Yao LQ,et al.Plasticity of ovarian cancer cell SKOV3ip and vasculogenic mimicry in vivo[J].Int J Gynecol Cancer,2008,18(3):476-486.
[37] Zhang S,Li M,Gu Y,et al.Thalidomide influences growth and vasculogenic mimicry channel formation in melanoma[J].J Exp Clin Cancer Res,2008,27:60.
[38] Fan YZ,Sun W,Zhang WZ,et al.Vasculogenic mimicry in human primary gallbladder carcinoma and clinical significance thereof[J].Zhonghua Yi Xue Za Zhi,2007,87(3):145- 149.
[39] Sun W,Fan YZ,Zhang WZ,et al.A pilot histomorphology and hemodynamic of vasculogenic mimicry in gallbladder carcinomas in vivo and in vitro[J].J Exp Clin Cancer Res,2011,30:46.
[40] Batmunkh E,Shimada M,Morine Y,et al.Expression of hypoxia-inducible factor-1 alpha (HIF-1 alpha) in patients with the gallbladder carcinoma[J].Int J Clin Oncol,2010,15(1):59-64.
[41] Wang CJ,Zhou ZG,Holmqvist A,et al.Survivin expression quantified by image pro-plus compared with visual assessment[J].Applied Immunohistochem Mol Morphol,2009,17(6):530-535.
[42] Xavier LL,Viola GG,Ferraz AC,et al.A simple and fast densitometric method for the analysis of tyrosine hydroxylase immunoreactivity in the substantia nigra pars compacta and in the ventral tegmental area[J].Brain Res Protocols,2005,16(1-3):58-64.
[43] Situ DR,Hu Y,Zhu ZH,et al.Prognostic relevance of beta-catenin expression in T2-3N0M0 esophageal squamous cell carcinoma[J].World J Gastroenterol,2010,16(41):5195-5202.
[44] Zlobec I,Steele R,Terracciano L,et al.Selecting immunohistochemical cut-off scores for novel biomarkers of progression and survival in colorectal cancer[J].J Clin Pathol,2007,60(10):1112-1116.
[45] Zhang S,Zhang D,Sun B.Vasculogenic mimicry:current status and future prospects[J].Cancer Lett,2007,254(2):157-164.
[46] Hendrix MJ,Seftor EA,Hess AR,et al.Molecular plasticity of human melanoma cells[J].Oncogene,2003,22(20):3070-3075.
[47] Folberg R,Arbieva Z,Moses J,et al.Tumor cell plasticity in uveal melanoma:microenvironment directed dampening of the invasive and metastatic genotype and phenotype accompanies the generation of vasculogenic mimicry patterns[J].Am J Pathol,2006,169(4):1376-1389.
[48] van der Schaft DW,Seftor RE,Seftor EA,et al.Effects of angiogenesis inhibitors on vascular network formation by human endothelial and melanoma cells[J].J Natl Cancer Inst,2004,96(19):1473-1477.
[49] Clarijs R,Otte-Holler I,Ruiter DJ,et al.Presence of a fluid-conducting meshwork in xenografted cutaneous and primary human uveal melanoma[J].Invest Ophthalmol Vis Sci,2002,43(4):912-918.
[50] Kobayashi H,Shirakawa K,Kawamoto S,et al.Rapid accumulation and internalization of radiolabeled herceptin in an inflammatory breast cancer xenograft with vasculogenic mimicry predicted by the contrast-enhanced dynamic MRI with the macromolecular contrast agent G6-(1B4M-Gd)(256)[J].Cancer Res,2002,62(3):860-866.
[51] Shirakawa K,Kobayashi H,Heike Y,et al.Hemodynamics in Vasculogenic mimicry and angiogenesis of inflammatory breast cancer xenograft[J].Cancer Res,2002,62(2):560-566.
[52] Shirakawa K,Kobayashi H,Sobajima J,et al.Inflammatory breast cancer:vasculogenic mimicry and its hemodynamics of an inflammatory breast cancer xenograft model[J].Breast Cancer Res,2003,5(3):136-139.
[53] Xu X,Jia R,Zhou Y,et al.Investigation of vasculogenic mimicry in sebaceous carcinoma of the eyelid[J].Acta Ophthalmol,2010,88(5):e160-164.
[54] Ahmadi SA,Moinfar M,Gohari Moghaddam K,et al.Practical application of angiogenesis and vasculogenic mimicry in prostatic adenocarcinoma[J].Arch Iran Med,2010,13(6):498-503.
[55] Postovit LM,Margaryan NV,Seftor EA,et al.Role of nodal signaling and the microenvironment underlying melanoma plasticity[J].Pigment Cell Melanoma Res,2008,21(3):348-357.
[56] Nadaoka J,Horikawa Y,Saito M,et al.Prognostic significance of HIF-1 alpha polymorphisms in transitional cell carcinoma of the bladder[J].Int J Cancer,2008,122(6):1297-1302.
[57] Guzman G,Cotler SJ,Lin AY,et al.A pilot study of vasculogenic mimicry immunohistochemical expression in hepatocellular carcinoma[J].Arch Pathol Lab Med,2007,131(12):1776-1781.
[58] Sun B,Zhang S,Zhao X,et al.Pilot study of molecular mechanism on vasculogenic mimicry in bi-directional differentiated malignant tumors[J].Chinese-German J Clin Oncol,2005,4(1):50-52.
[59] Hillen F,Griffioen AW.Tumour vascularization:sprouting angiogenesis and beyond[J].Cancer Metastasis Rev,2007,26(3-4):489-502.
[60] Mukhopadhyay D,Datta K.Multiple regulatory pathways of vascular permeability factor/vascular endothelial growth factor (VPF/VEGF) expression in tumors[J].Seminars in Cancer Biol,2004,14(2):123-130.
[61] 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.
[62] Seftor RE,Seftor EA,Kirschmann DA,et al.Targeting the tumor microenvironment with chemically modified tetracyclines:inhibition of laminin 5 gamma2 chain promigratory fragments and vasculogenic mimicry[J].Mol Cancer Ther,2002,1(13):1173-1179.
[63] Seftor RE,Seftor EA,Koshikawa N,et al.Cooperative interactions of laminin 5 gamma2 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.