Legumain在葡萄膜黑色素瘤中的表达及其对患者预后影响的研究
摘要
[目的]葡萄膜黑色素瘤是一种成人最常见的原发性眼内恶性肿瘤,主要来源于葡萄膜黑色素细胞,因其血运丰富,主要经血行转移,而且特异性转移到肝脏,转移是主要的致死因素,尤其葡萄膜黑色素瘤的转移往往具有隐匿性,即在肿瘤早期就容易出现临床影像学无法诊断发现的微转移灶。因而目前对葡萄膜黑色素瘤研究的热点是确立转移潜能的预后标志,寻找可以早期准确客观预判葡萄膜黑色素瘤患者预后的生物标记物。legumain是一个新发现的半胱氨酸蛋白酶,其特点：在肿瘤组织中特异性表达,正常组织中无明显表达,与肿瘤免疫关系密切,而且与肿瘤生成,侵袭,转移,扩散密切相关。本研究旨在明确legumain在葡萄膜黑色素瘤中的表达,探讨legumain对葡萄膜黑色素瘤患者预后的影响,及其与影响葡萄膜黑色素瘤患者预后各因素(病理类型,肿瘤体积,转移情况,视神经受累,侵润程度)的关系。
[方法]本研究首先选取两株葡萄膜黑色素瘤细胞株Mum-2B和Mum-2C,进行细胞培养传代,提取RNA和蛋白质,进行RT-PCR和Western-blot检测,同时利用两株细胞系进行细胞免疫荧光的染色,从而在基因和蛋白水平明确legumain在葡萄膜黑色素瘤细胞系的表达。在这个实验基础上,进而收集82例葡萄膜黑色素瘤患者的临床及预后资料,同时将这些患者保存的石蜡标本制作组织切片,进行legumain免疫组织化学染色,统计legumain的表达情况,运用Kaplan-meier单因素生存分析的原理与方法进行统计分析,将legumain的表达情况作为待检验的预后因素,与患者预后结果相联系,并运用Log-rank、Brelow、Tarone-Ware三种方法进行显著性检验,明确legumain对葡萄膜黑色素瘤患者预后的影响,同时对82例葡萄膜黑色素瘤的临床预后资料进行Kaplan-meier单因素生存分析,并运用Log-rank、Brelow、Tarone-Ware三种方法进行显著性检验,筛选出与葡萄膜黑色素瘤患者预后相关的因素,然后将legumain免疫组化结果分别与葡萄膜黑色素瘤患者预后相关的各因素进行相关分析,明确legumain与影响葡萄膜黑色素瘤预后各相关因素的关系。
[结果]在细胞实验中,通过RT-PCR的检测,表明在基因水平上两株葡萄膜黑色素瘤细胞系legumain为高表达,从细胞免疫荧光也可见legumain在两株葡萄膜黑色素瘤细胞系细胞膜呈现为绿色荧光的高表达,利用western-blot技术对两株细胞系进行legumain的蛋白检测,可见legumain在两株细胞系中均为高表达,且在Mum-2B中表达较高。对82例葡萄膜黑色素瘤患者的石蜡病理切片进行免疫组化染色,结果显示在葡萄膜黑色素瘤的三种基本病理类型(上皮细胞型,梭形细胞型,混合细胞型)中legumain均有阳性(高表达)和阴性(低表达)的表达。通过Kaplan-meier单因素生存分析和Log-rank、Brelow、Tarone-Ware三种方法进行显著性检验,发现葡萄膜黑色素瘤患者性别的不同对患者预后无明显影响(P>0.05),其余因素legumain,病理类型,肿瘤体积,转移情况,视神经受累,侵润程度均对葡萄膜黑色素瘤患者的预后有显著性的影响(P<0.05)。经过双变量的相关分析,legumain与影响葡萄膜黑色素瘤患者预后的各因素(病理类型,肿瘤体积,转移情况,侵袭视神经,侵润程度)均呈正相关(P<0.01),而且与肿瘤侵润程度关系密切(相关系数大于0.5)。
[结论]本研究通过对葡萄膜黑色素瘤细胞系和石蜡包埋病理标本进行legumain表达情况的检测,发现legumain在葡萄膜黑色素瘤中有明显的高表达,然后结合患者临床及预后资料进行葡萄膜黑色素瘤的预后研究和相关分析,明确legumain与葡萄膜黑色素瘤预后及影响预后各因素的关系,发现legumain不但与葡萄膜黑色素瘤患者预后有关,而且与影响葡萄膜黑色素瘤患者预后的各因素有一定的相关性,从而说明legumain不仅是判断葡萄膜黑色素瘤患者预后的一个重要指标,而且在葡萄膜黑色素瘤的发生发展过程中可能起着重要作用,legumain也有可能成为葡萄膜黑色素瘤治疗的一个新的靶点。
[objective] In adults, uveal melanoma is the most common malignant intraocular tumor, which originated from melanocytes. Because of its rich circulation of blood, uveal melanoma can disseminate hematogenously, which preferentially metastasizes to the liver, and metastasis is the main cause of death. Especially insidious potential is the main character of UM metastasis, so at the time of diagnosis, many patients may have subclinical metastasis, which underscores the critical need to identify prognostic markers indicative of uveal melanoma invasive metastatic potential. Legumain, an novel asparaginyl endopeptidase, is expressed in some humam solid carcinoma, whereas it can not been detected in normal tissures, whose overexpression has been shown to be combined with increased migration, invasion, and metastasis in some solid carcinaoma. In our study, to make sure the expression of legumain in uveal melanoma, and to investigate its prognostic value in the patients of uveal melanoma and its relationship with other prognostic factors (cell type, tumor size, metastasis, involvement of optic nerve, and extent of invasion) in uveal melanoma.
[Methods] In our study, firstly, to detect legumain in two uveal melanoma cell lines by using RT-PCR and western-blot, after the culture of cell lines and extraction of RNA and protein from cell lines. Simultaneously, immunofluorescence staining can be used to show the expression of legumain in uveal melanoma cell lines. Secondly, Paraffin-embedded sections from 82 clinicopathologically well-characterized cases of primary uveal melanoma were immunohistochemically stained for legumain. Legumain expression was evaluated in terms of both the intensity and the extent of staining for each specimen. Thirdly, to investigate the relationship between the expression of legumain and prognostic outcome of uveal melanoma patients through Kaplan-Meier univariate analysis and Log-rank, Brelow, Tarone-Ware tests, and some other prognostic risk factors were to be determined after the prognostic risk factors wre preliminary srcreened respectively with Kaplan-Meier univariate analysis and Log-rank, Brelow, Tarone-Ware tests. Last but not least, we investigated the relationship between the expression of legumain and other prognostic risk factors through bivariate correlation analysis.
[Results] In our experiment, the expression of legumain was detected in two uveal melanoma cell lines by using RT-PCR, western-blot and immunofluorescence staining, and especially the expression of legumain was more obvious in Mum-2B cell line by western-blot. Both positive and negative expression of legumain can be shown in the three main cell types of uveal melanoma (epitheloid, spindle, mixed cell) by immunohistochemically staining for paraffin-embedded sections from 82 cases of uveal melanoma. Through Kaplan-Meier univariate analysis and Log-rank, Brelow, Tarone-Ware tests, the expression of legumain, cell type, tumor size, metastasis, involvement of optic nerve, and extent of invasion (P<0.05) except sex (P>0.05) had not only great influence on the prognosis of uveal melanoma patients, but also the expression of legumain had positive correlations with other prognostic risk factors including cell type, tumor size, metastasis, involvement of optic nerve, and extent of invasion (P<0.01), especially with the extent of invasion (correlation coefficient> 0.5), by using bivariate correlation analysis.
[Conclusion] In this study, by detecting legumain in the two cell lines of uveal melanomna and paraffin-embedded sections from 82 cases of uveal melanoma, it can be proved that legumain was expressed in uveal melanoma, and in combination with clinical data, through survival study and correlation analysis, it is to find that upregulation of legumain expression appears to be associated with poor prognosis in uveal melanoma, and also has positive correlations with other prognostic risk factors. Therefore, legumain is not only a valuable predictive marker for prognosis of uveal melanoma patients, but also it is involved in the pathgenesis and progress of uveal melanoma. In the near future, legumain may be the important target for effective treatment of uveal melanoma.
[方法]本研究首先选取两株葡萄膜黑色素瘤细胞株Mum-2B和Mum-2C,进行细胞培养传代,提取RNA和蛋白质,进行RT-PCR和Western-blot检测,同时利用两株细胞系进行细胞免疫荧光的染色,从而在基因和蛋白水平明确legumain在葡萄膜黑色素瘤细胞系的表达。在这个实验基础上,进而收集82例葡萄膜黑色素瘤患者的临床及预后资料,同时将这些患者保存的石蜡标本制作组织切片,进行legumain免疫组织化学染色,统计legumain的表达情况,运用Kaplan-meier单因素生存分析的原理与方法进行统计分析,将legumain的表达情况作为待检验的预后因素,与患者预后结果相联系,并运用Log-rank、Brelow、Tarone-Ware三种方法进行显著性检验,明确legumain对葡萄膜黑色素瘤患者预后的影响,同时对82例葡萄膜黑色素瘤的临床预后资料进行Kaplan-meier单因素生存分析,并运用Log-rank、Brelow、Tarone-Ware三种方法进行显著性检验,筛选出与葡萄膜黑色素瘤患者预后相关的因素,然后将legumain免疫组化结果分别与葡萄膜黑色素瘤患者预后相关的各因素进行相关分析,明确legumain与影响葡萄膜黑色素瘤预后各相关因素的关系。
[结果]在细胞实验中,通过RT-PCR的检测,表明在基因水平上两株葡萄膜黑色素瘤细胞系legumain为高表达,从细胞免疫荧光也可见legumain在两株葡萄膜黑色素瘤细胞系细胞膜呈现为绿色荧光的高表达,利用western-blot技术对两株细胞系进行legumain的蛋白检测,可见legumain在两株细胞系中均为高表达,且在Mum-2B中表达较高。对82例葡萄膜黑色素瘤患者的石蜡病理切片进行免疫组化染色,结果显示在葡萄膜黑色素瘤的三种基本病理类型(上皮细胞型,梭形细胞型,混合细胞型)中legumain均有阳性(高表达)和阴性(低表达)的表达。通过Kaplan-meier单因素生存分析和Log-rank、Brelow、Tarone-Ware三种方法进行显著性检验,发现葡萄膜黑色素瘤患者性别的不同对患者预后无明显影响(P>0.05),其余因素legumain,病理类型,肿瘤体积,转移情况,视神经受累,侵润程度均对葡萄膜黑色素瘤患者的预后有显著性的影响(P<0.05)。经过双变量的相关分析,legumain与影响葡萄膜黑色素瘤患者预后的各因素(病理类型,肿瘤体积,转移情况,侵袭视神经,侵润程度)均呈正相关(P<0.01),而且与肿瘤侵润程度关系密切(相关系数大于0.5)。
[结论]本研究通过对葡萄膜黑色素瘤细胞系和石蜡包埋病理标本进行legumain表达情况的检测,发现legumain在葡萄膜黑色素瘤中有明显的高表达,然后结合患者临床及预后资料进行葡萄膜黑色素瘤的预后研究和相关分析,明确legumain与葡萄膜黑色素瘤预后及影响预后各因素的关系,发现legumain不但与葡萄膜黑色素瘤患者预后有关,而且与影响葡萄膜黑色素瘤患者预后的各因素有一定的相关性,从而说明legumain不仅是判断葡萄膜黑色素瘤患者预后的一个重要指标,而且在葡萄膜黑色素瘤的发生发展过程中可能起着重要作用,legumain也有可能成为葡萄膜黑色素瘤治疗的一个新的靶点。
[objective] In adults, uveal melanoma is the most common malignant intraocular tumor, which originated from melanocytes. Because of its rich circulation of blood, uveal melanoma can disseminate hematogenously, which preferentially metastasizes to the liver, and metastasis is the main cause of death. Especially insidious potential is the main character of UM metastasis, so at the time of diagnosis, many patients may have subclinical metastasis, which underscores the critical need to identify prognostic markers indicative of uveal melanoma invasive metastatic potential. Legumain, an novel asparaginyl endopeptidase, is expressed in some humam solid carcinoma, whereas it can not been detected in normal tissures, whose overexpression has been shown to be combined with increased migration, invasion, and metastasis in some solid carcinaoma. In our study, to make sure the expression of legumain in uveal melanoma, and to investigate its prognostic value in the patients of uveal melanoma and its relationship with other prognostic factors (cell type, tumor size, metastasis, involvement of optic nerve, and extent of invasion) in uveal melanoma.
[Methods] In our study, firstly, to detect legumain in two uveal melanoma cell lines by using RT-PCR and western-blot, after the culture of cell lines and extraction of RNA and protein from cell lines. Simultaneously, immunofluorescence staining can be used to show the expression of legumain in uveal melanoma cell lines. Secondly, Paraffin-embedded sections from 82 clinicopathologically well-characterized cases of primary uveal melanoma were immunohistochemically stained for legumain. Legumain expression was evaluated in terms of both the intensity and the extent of staining for each specimen. Thirdly, to investigate the relationship between the expression of legumain and prognostic outcome of uveal melanoma patients through Kaplan-Meier univariate analysis and Log-rank, Brelow, Tarone-Ware tests, and some other prognostic risk factors were to be determined after the prognostic risk factors wre preliminary srcreened respectively with Kaplan-Meier univariate analysis and Log-rank, Brelow, Tarone-Ware tests. Last but not least, we investigated the relationship between the expression of legumain and other prognostic risk factors through bivariate correlation analysis.
[Results] In our experiment, the expression of legumain was detected in two uveal melanoma cell lines by using RT-PCR, western-blot and immunofluorescence staining, and especially the expression of legumain was more obvious in Mum-2B cell line by western-blot. Both positive and negative expression of legumain can be shown in the three main cell types of uveal melanoma (epitheloid, spindle, mixed cell) by immunohistochemically staining for paraffin-embedded sections from 82 cases of uveal melanoma. Through Kaplan-Meier univariate analysis and Log-rank, Brelow, Tarone-Ware tests, the expression of legumain, cell type, tumor size, metastasis, involvement of optic nerve, and extent of invasion (P<0.05) except sex (P>0.05) had not only great influence on the prognosis of uveal melanoma patients, but also the expression of legumain had positive correlations with other prognostic risk factors including cell type, tumor size, metastasis, involvement of optic nerve, and extent of invasion (P<0.01), especially with the extent of invasion (correlation coefficient> 0.5), by using bivariate correlation analysis.
[Conclusion] In this study, by detecting legumain in the two cell lines of uveal melanomna and paraffin-embedded sections from 82 cases of uveal melanoma, it can be proved that legumain was expressed in uveal melanoma, and in combination with clinical data, through survival study and correlation analysis, it is to find that upregulation of legumain expression appears to be associated with poor prognosis in uveal melanoma, and also has positive correlations with other prognostic risk factors. Therefore, legumain is not only a valuable predictive marker for prognosis of uveal melanoma patients, but also it is involved in the pathgenesis and progress of uveal melanoma. In the near future, legumain may be the important target for effective treatment of uveal melanoma.
引文
[1]Folberg R. Tumor progression in ocular melanomas. J Invest Dermatol 1993; 100:1389-98.
[2]Zimmerman L, McLean I. Do growth and onset of symptoms of uveal melanomas indicate subclinical metastasis? Ophthalmology 1984; 92:685-91.
[3]Donoso L, Shields J, Augsburger J et al. Metastatic uveal melanoma;Diffuse hepatic metastasis in a patient with concurrent normal serum enzyme levels and liver scan (letter). Arch Ophthalmol 1985; 103:758.
[4]Egan KM, Seddon JM, Glym RJ, et al. Edidemiologic aspects of uveal melanoma. Sur Ophthalmol,1988,32:239-251
[5]Mooy C, De Jong P. Prognostic parameters in uveal melanoma:A review. Surv Ophthalmol 1996; 41:215-28.
[6]李鹏程,张虹,丁正平,等TGF β RII和Smad4蛋白在葡萄膜黑色素瘤中的表达[J].肿瘤防治研究,2005,32(1)：35-36.
[7]吴中耀,郭琳洁,郑湖玲.葡萄膜黑色素瘤转移相关基因的表达及意义[J].中国实用眼科杂志,2001,19(2)：342-345.
[8]Al-Jamal RT, Makitie T, Kivela T. Nucleolar diameter and microvascular factors as independent predictors of mortality from malignant melanoma of the choroid and ciliary body. Invest Ophthalmol Vis Sci 2003; 44:2381-2389.
[9]郭琳洁,吴中耀,郑湖玲.细胞周期相关基因在葡萄膜黑色素瘤中表达与意义[J].中华眼底病杂志,2001,17(1)：44-46.
[10]傅涛,魏文斌,王阳.血清黑色素瘤活动抑制蛋白与葡萄膜黑色素瘤[J].中华眼底病杂志,2005,21(1)：153-155.
[11]陈陆霞,张诗武,孙保存.MMP22、MMP29的表达与脉络膜黑色素瘤微血管密度及肿瘤转移的关系[J].眼科新进展,2007,8(27)：565-567
[12]Chen JM, Dando PM, Rawlings ND, et al. Cloning, isolation, and characterization of mammalian legumain, an asparaginyl endopeptidase. Biol Chem 1997;272:8090-8.
[13]Jessica G, Frank T, Hans J. Legumain expression as a prognostic factor in breast cancer patients. Breast Cancer Res Treat 2007; 102:1-6
[14]Raghavendra V M, Gunnar A, Jingfang G. Legumain Expression in Relation to Clinicopathologic and Biological Variables in Colorectal Cancer. Clinical Cancer Research 2005; 11:2293-2299.
[15]Watts C, Matthews SP, Mazzeo D, et al. Asparaginyl endopeptidase:case history of a class ⅡMHC compartment p rotease [J]. Immunol Rev,2005,207:218-228.
[16]Luo Y, Zhou H, Krueger J, et al. Targeting tumor-associated macrophages as a novel strategy against breast cancer [J]. J Clin Invest,2006,116 (8):2132-2141.
[17]孙为荣.眼科病理学[M].北京：人民卫生出版社,1997.266-280.
[18]Zimmerman L, McLean I. Do growth and onset of symptoms of uveal melanomas indicate subclinical metastasis? Ophthalmology 1984; 92:685-91.
[19]Donoso L, Shields J, Augsburger J et al. Metastatic uveal melanoma;Diffuse hepatic metastasis in a patient with concurrent normal serum enzyme levels and liver scan (letter). Arch Ophthalmol 1985; 103:758.
[20]Chen JM, Dando PM, Rawlings ND, et al. Cloning, isolation, and characterization of mammalian legumain, an asparaginyl endopeptidase. Biol Chem 1997;272:8090-8.
[21]Watts C, Matthews SP, Mazzeo D, et al. Asparaginyl endopeptidase:case history of a class Ⅱ MHC compartment p rotease [J]. Immunol Rev,2005,207:218-228.
[22]Manoury, B., Hewitt, E. W., Morrice, N., Dando, P. M., Barrett, A. J., and Watts, C.An asparaginyl endopeptidase processes a microbial antigen for class II MHC presentation. Nature,396:695-699,1998.
[23]Choi, S. J., Kurihara, N., Oba, Y., and Roodman, G. D. Osteoclast inhibitory peptide inhibits osteoclast formation via its C-terminal fragment. J. Bone Miner. Res., 16:1804-1811,2001.
[24]Cheng L, Chengzao S, Haining H, Kim J, et al Overexpression of legumain in tumorsis significant for invasion/metastasis and a candidate enzymatic target for prodrug therapy. Cancer Res 2003; 63:2957-2964,
[25]Chen JM, Dando PM, Stevens RA. Cloning and expression of mouse legumain, a lysosmal endopeptidase. Biochem J,1988,335:111-117.
[26]Albini A, Iwamoto Y, Kleinman HK, et al. A rapid in vitro assay for quantitating the invasive potential of tumor cells. Cancer Res,1987,47(12):3239 —3245.
[27]吴文苑,余涟,刘春,罗金刚Legumain对肿瘤细胞侵润力和血管内皮细胞管腔形成的作用.热带医学杂志,2009,9(8)：864-867
[28]Susanna L, He Z, Huai-dong H, et al A Legumain-based minigene vaccine targets the tumor stromaand suppresses breast cancer growth and angiogenesis Cancer Immunol Immunother (2008) 57:507-515
[29]Luo Y, Zhou H, Krueger J, Kaplan C, Lee SH, Dolman C, Markowitz D, Wu W, Liu C, Reisfeld RA, Xiang R. Targeting tumor-associated macrophages as a novel strategy against breast cancer. J Clin Invest 2006; 116:2132
[30]Jedeszko C, Sloane BF. Cysteine cathepsins in human cancer. Biol Chem,2004, 385 (11):1017-1027.
[31]Yoshikata M, Hisazumi A, Toshiro S, et al. Legumain/asparaginyl endopeptidase controls extracellular matrix remodeling through the degradation of fibronectin in mouse renal proximal tubular cells FEBS Lett.2007,581 (7):1417-1424.
[32]Bingle L, Brown NJ, Lewis CE. The role of tumor-associated macrophages in tumor progression:imp lications for new anticancer therapies[J]. J Pathol,2002, 196 (3):254-256.
[33]Lewis CE, Pollard JW. Distinct role of macrophages in different tumor micro-environments[J]. Cancer Res,2006,66 (2):605-612.
[34]郭强,李康,徐薇,储以微,熊思东.Legumain在荷瘤与正常小鼠脾细胞中的差异性表达.中国癌症杂志,2008,18(2)：101-105
[35]Wolk K, Grutz G, Witte K, et al. The exp ression of Legumain, an asparaginyl endopep tidase that controls antigen p rocessing, is reduced in endotoxin-tolerant monocytes [J]. Genes Immun,2005,6 (5):452-456.
[36]Luo Y, Zhou H, Liu C, et al. A novel stress protein, legumain, is a target for a genetic vaccine against breast cancer [A]. AACR Meeting Abstracts [C],2005,5 (1):1216-1217.
[37]Elisabeth AS, Paul SM, Dawn AK, Jacob P, Andrew JM, Jeffrey MT, Robert F& Mary J.C. Hendrix. Molecular determinants of human uveal melanoma invasion and metastasis. Clinical & Experimental Metastasis 19:233-246,2002.
[38]颜建华,吴中耀.葡萄膜肿瘤.见：吴中耀,主编.现代眼肿瘤眼眶病学.北京：人民军医出版社,2002.222-232.
[39]王风华,李彬.脉络膜黑色素瘤的临床、组织病理学特点及研究进展.国外医学眼科学分册,2001,25：372-377.
[40]McLean IW, Foster WD, Zimmerman L E, et al. Modifications of Callender's classification of uveal melanoma at the Armed Forces Institute of Pathology. Am J Ophthalmol,1983,96:502-509.
[41]梁建宏,易长贤,Schilling H,等.脉络膜恶性黑色素瘤中Ki267的表达和肿瘤主要组织病理特性的关系.眼科学报,2001,17：114-117.
[42]林锦镛.葡萄膜黑色素瘤的临床病理学特征和鉴别诊断要点.中华眼科杂志,2003,39：121-124.
[43]罗清礼,孙宪丽.努力提高脉络膜黑色素瘤的基础和临床研究水平.中华眼科杂志,2003,39：65267.
[44]Coleman K, Baak J A, Van Diest P, Mullaney J, Farrell M & Fenton M Prognostic factors following enucleation of 111 uveal melanomas. Br J Ophthalmol 1993,77:688-692
[45]Shields J A (1992):Posterior uveal melanoma:clinical and pathological features. In:Shields J A & Shields C L (eds). Intraocular tumors. A text and atlas. WB Saunders Co, Philadelphia.
[46]Kujala E, Makitie T, Kivela T. Very long-term prognosis of patients with malignant uveal melanoma. Invest Ophthalmol Vis Sci 2003;44:4651-9.
[47]Scholes AG, Damato BE, Nunn J, Hiscott P, Grierson I & Field JK:Monosomy 3 in uveal melanoma:correlation with clinical and histologic predictors of survival. Invest Ophthalmol Vis Sci 2003,44:1008-1011.
[48]Makitie T, Summanen P, Tarkkanen A &Kivela T:Microvascular density in predicting survival of patients with choroidal and ciliary body melanoma. Invest Ophthalmol Vis Sci 1999,40:2471-2480.
[49]Seregard S, Kock E. Prognostic indicators following enucleation for posterior uveal melanoma:a multivariate analysis of long-term survival with minimized loss to follow-up. Acta Ophthalmol Scand 1995;73:340-4.
[50]郭琳洁,吴中耀,张胜,陈家祺,艾思明,郑湖玲.葡萄膜黑色素瘤病理学 特征与预后的关系.中华病理学杂志,2002,31(6)：518-522
[51]Sarah E, Ian C, Bertil D. Routes of Extraocular Extension of Uveal Melanoma Ophthalmology.2008; 115 (10):1778-1785
[52]Patel KA, Edmondson ND, Talbot F,Parsons MA, Rennie IG & Sisley K Prediction of prognosis in patients with uveal melanoma using fluorescence in situ hybridization. Br J Ophthalmol 2001,85:1440-1444.
[53]Haddab S, Hidayat A, Tabbara KF. Ciliary body melanoma with optic nerve invasion. Br J Ophthalmol.1990;74:123-124.
[54]Chess J, Albert DM, Bellows AR, Dallow R. Uveal melanoma:case report of extension through the optic nerve to the surgical margin in the orbital apex. Br J Ophthalmol.1984;68:272-275.
[55]Abdel-Rahman MH, Craig EL, Davidorf FH, Eng C. Expression of vascular endothelial growth factor in uveal melanoma is independent of 6p21-region copy number. Clin Cancer Res.2005; 11:73-78.
[56]Spencer WH. Optic nerve extension of intraocular neoplasms. Am J Ophthalmol. 1975;80:465-471.
[57]Walker TM, Van Ginkel PR, Gee RL, et al. Expression of angiogenic factors Cyr61 and tissue factor in uveal melanoma. Arch Ophthalmol.2002;120:1719-1725.
[58]Boyd SR, Tan DS, de Souza L, et al. Uveal melanomas express vascular endothelial growth factor and basic fibroblast growth factor and support endothelial cell growth. Br J Ophthalmol.2002;86:440-447.
[59]Cohn-Cedermark G, Mansson-Brahme E, Rutqvist LE, Larsson O, Johansson H, Ringborg U. Central nervous system metastases of cutaneous malignant melanoma—a population-based study. Acta Oncol.1998;37:463-470.
[60]Donoso LA, Nagy RM, Brockman RJ, et al. Metastatic uveal melanoma. Hepatic cell surface enzymes, isoenzymes, and serum sialic acid levels in early metastatic disease.Arch Ophthalmol.1983;101:791-794.
[61]Schaller UC, Bosserhoff AK, Neubauer AS, et al. Melanoma inhibitory activity: a novel serum marker for uveal melanoma. Melanoma Res.2002; 12:593-599.
[62]Maniotis AJ, Folberg R, Hess A, et al. Vascular channel formation by human melanoma cells in vivo and in vitro:vasculogenic mimicry. Am J Pathol,1999, 155:739-752
[63]Gawenda J, Traub F, Luck HJ, et al. Legumain expression as a prognostic factor in breast cancer patients. Breast Cancer Res Treat.2007,102 (1):126.
[64]Murthy RV, Arbman G, Gao J, Roodman GD, Sun XF. Legumain expression in relation to clinicopathologic and biological variables in colorectal cancer. Clin Cancer Res 2005; 11:2293
[65]Beck, H., Schwarz, G., Schroter, C. J., Deeg, M., Baier, D., Stevanovic, S., Weber,E., Driessen, C., and Kalbacher, H. Cathepsin S and an asparagine-specific endoprotease dominate the proteolytic processing of human myelin basic protein in vitro. Eur. J. Immunol.,2001.31:3726-3736,
[66]Wu W, Luo Y, Sun C, et al. Targeting cell-impermeable prodrug activation to tumor microenvironment eradicates multiple drug-resistant neoplasms [J]. Cancer Res,2006,66 (2):970-980.
[1]Sobin LH, Wittekind Ch. TNM Classification of Malignant Tumours. New York: Wiley-Liss; 2002.
[2]Lorigan JG, Wallace S, Mavligit GM. The prevalence and location of metastases from ocular melanoma:imaging study in 110 patients. AJR Am J Roentgenol. 1991;157:1279-
[3]Aoyama T, Mastrangelo MJ, Berd D, et al. Protracted survival after resection of metastatic uveal melanoma. Cancer.2000;89:1561-1568.
[4]Eskelin S, Pyrhonen S, Summanen P, et al. Screening for metastatic malignant melanoma of the uvea revisited. Cancer.1999;85:1151-1159.
[5]Bergman L, Seregard S, Nilsson B, et al. Uveal melanoma survival in Sweden from 1960 to 1998. Invest Ophthalmol Vis Sci.2003;44:3282-3287.
[6]Kujala E, Ma T, Kivela T. Very long-term prognosis of patients with malignant uveal melanoma. Invest Ophthalmol Vis Sci.2003;44:4651-4659.1281.
[7]Collins VP, Loeffler RK, Tivey H. Observations on growth rates of human tumors. AJR Am J Roentgenol.1956;76:988-1000.
[8]Eskelin S, Pyrho S, Summanen P, et al. Tumor doubling times in metastatic malignant melanoma of the uvea:tumor progression before and after treatment. Ophthalmology.2000; 107:1443-1449.
[9]Singh AD. Uveal melanoma:implications of tumor doubling time [Letter]. Ophthalmology.2001;108:829-831.
[10]Manschot WA, van Peperzeel HA. Choroidal melanoma. Enucleation or observation? A new approach. Arch Ophthalmol.1980;98:71-77.
[11]Patel JK, Didolkar MS, Pickren JW, et al. Metastatic pattern of malignant melanoma. A study of 216 autopsy cases. Am J Surg.1978;135:807-810.
[12]Gombos DS, Van Quill KR, Uusitalo M, et al. Geographic disparities in diagnostic screening for metastatic uveal melanoma. Ophthalmology.2004; 111:2254-2258.
[13]Diener-West M, Reynolds SM, Agugliaro DJ, et al. Screening for metastasis from choroidal melanoma:the Collaborative Ocular Melanoma Study Group Report 23. J Clin Oncol.2004;22:2438-2444.
[14]Eskelin S, Pyrho nen S, Hahka-Kemppinen M, et al. A prognostic model and staging for metastatic uveal melanoma. Cancer.2003;97:465-475.
[15]Kaiserman I, Amer R. Pe'er J. Liver function tests in metastatic uveal melanoma. Am J Ophthalmol.2004;137:236-243.
[16]Eskelin S, Pyrhonen S, Summanen P, et al. Screening for metastatic malignant melanoma of the uvea revisited. Cancer.1999;85:1151-1159.
[17]The Collaborative Ocular Melanoma Study Group. Assessment of metastatic diseasestatus at death in 435 patients with large choroidal melanoma in the Collaborative Ocular Melanoma Study (COMS):COMS report no.15. Arch Ophthalmol.2001;119:670-676.
[18]The Collaborative Ocular Melanoma Study Group. The Collaborative Ocular Melanoma Study (COMS) randomized trial of pre-enucleation radiation of large choroidal melanoma Ⅰ:characteristics of patients enrolled and not enrolled. COMS report no.9. Am J Ophthalmol.1998;125:767-778.
[19]Missotten GS, Tang NE, Korse CM, et al. Prognostic value of S-100-beta serum concentration in patients with uveal melanoma. Arch Ophthalmol.2003; 121:1117 1119.
[20]Schaller UC, Bosserhoff AK, Neubauer AS, et al. Melanoma inhibitory activity: a novel serum marker for uveal melanoma. Melanoma Res.2002;12:593-599.
[21]Donoso LA, Berd D, Augsburger JJ, et al. Metastatic uveal melanoma. Pretherapy serum liver enzyme and liver scan abnormalities. Arch Ophthalmol. 1985;103:796-798.
[22]Donoso LA, Nagy RM, Brockman RJ, et al. Metastatic uveal melanoma. Hepatic cellsurface enzymes, isoenzymes, and serum sialic acid levels in early metastatic disease. Arch Ophthalmol.1983;101:791-794.
[23]Diener-West M, Earle JD, Fine SL, et al. The COMS randomized trial of iodine
125 brachytherapy for choroidal melanoma, Ⅱ:characteristics of patients enrolled and not enrolled. COMS report no.17. Arch Ophthalmol.2001; 119:951-965.
[24]Myatt N, Aristodemou P, Neale MH, et al. Abnormalities of the transforming growth factor-beta pathway in ocular melanoma [J]. J Pathol,2000,192:511-518
[25]Sun L. Tumor-supp ressive and p romoting function of transforming growth factor beta [J]. Front Biosci,2004,9:1925-1935
[26]Ramont L, Pasco S, Hornebeck W, et al. Transforming growth factor-betal inhibits tumor growth in a mouse melanoma model by down-regulating the plasminogen activation system[J]. Exp Cell Res,2003,291: 1-10
[27]Park SS,Li L, Korn TS, et al. Effect of transforming growth factor2beta on plasminogen activator p roduction of cultured human uvealmelanoma cells [J]. Curr Eye Res,1996,15:755-763
[28]Ijland SA, Jager MJ, Heijdra BM, et al. Exp ression of angiogenic and immunosuppressive factors by uveal melanoma cell lines [J]. Melanoma Res,1999, 9:445-450
[29]EI-Shabrawi Y, Ardjomand N, Radner H, et al MMP-9 is predominantly expressed in epithelioid and not spindle cell uveal melanoma. J Pathol 2001; 194 (3): 201-206
[30]Foda H, Zucker S. Matrix metallop roteinases in cancer invasion metastasis and angiogenesis. Drug Discov Today 2001; 6:478-482
[31]Garbell EA, ReedMW, Brown NJ, et a.l Proleolysis in human breast and colorectal cancer. Br J Cancer 1999; 81 (2):287-293
[32]Gilles C, PoletteM, Coraux C. Contribution ofMT12MMP and of human laminin25 gamma22 chain degradation to mammary epithelial cell migration. J Cell Scil 2001; 11 (4):2967-2976
[33]Hotary K, Allen E, Puntrieri A. Regulation of cell invasion and morphogenesis in a three-dimensional type I collagen matrix by membrane type metallop roteinase 1,2 and 3. J Cell Biol 2000; 149 (6):1309-1323
[34]Baker JK, Elshaw SR, Mathewman GE, et a.l Expression of integrins, degradeative enzymes and their inhibitors in uveal melanoma:differences between invitro and invivo expression. Melan Res 2001; 11(3):265-273
[35]Broooks PC, Silllettl S, Von Schalscha TL. Disruption of angiogenesis by PEX, a noncatalytic metalloproteinase fragment thintegring binding cativity. Cell 1998; 92 (3):391-400
[36]Fang J, Shing Y, Wiederschaim D, et a.l Matrix Matallop roteinase-2 is required for the switch to the angiogenic phenol-type in a tumor model. ProcNatlA cad Sci USA 2000; 1997 (8):3884-3889
[37]Vannen A, Kallioinen M, von Dickh off K, et a.l. Matrix metallop roteinase-2 (MMP-2) immunoreactive protein a new prognostic marker in uveal melanoma. J Pathol 1999; 188 (1):56-62
[38]陈陆霞,张诗武,孙保存,等.MMP22、MMP29的表达与脉络膜黑色素瘤微血管密度及肿瘤转移的关系.眼科新进展2007；27(8)：565-567
[39]栗映梅,申家泉.基质金属蛋白酶及其组织抑制因子与眼病的相关性研究进展.国际眼科杂志2007；7(4)：1135-1138
[40]陈翔,田立红,谢红付,等.CD147对成纤维细胞MMP29的产生和恶性黑色素瘤细胞浸润的作用.中南大学学报(医学版)2005；30(3)：249-252
[41]Maniotis AJ, Folberg R, Hess A, et al. Vascular channel formation by human melanoma cells in vivo and in vit ro:vasculogenic mimicry. Am J Pathol,1999, 155:739-752
[42]Folberg R, Pe'er J, Gruman LM, et al. The morphologic characteristics of tumor blood vessels as a marker of tumor progression in primary human uveal melanoma:a matched case2control study. Hum Pathol,1992,23:1298-1305
[43]Hendrix MJ, Seftor EA, Meltzer PS, et al. Expression and functional significance of VE-cadherin in aggressive human melanoma cells:role in vasculogenic mimicry. Proc Natl Acad Sci USA,2001,98:8018-8023
[44]Bittner M, Meltzer P, Chen Y, et al. Molecular classification of cutaneous malignant melanoma by gene expression profiling. Nature,2000,406:536-540
[45]Hendrix MJ, Seftor EA, Seftor RE, et al Regulation of uveal melanoma intercoverted phenotype by hepatocyte growth factor(HGF). Am J Pathol 1998; 152: 855-863
[46]Hendrix MJ, Seftor EA, Seftor RE, et al Biologic determinants of uveal melanoma metastatic phenotype role of intermediate filaments as predictive markers. Lab Invest 1998; 78:153-163
[47]Malikarjuna K, Pushpara V, Biswas J, et al Expression of epidermal growth factor receptor, ezin, hepatocyte growth factor and c-Met in uveal melanoma an immunohistochemical study. Curr Eye Res 2007; 32:281-90
[48]Cesare S, Marshall JC, Logan P et al Expression and migratory analysis of 5 human uveal melanoma cell lines for CXCL12, CXCL8, CXCL1, and HGF. J Carcinog 2007,6:34-42
[49]Woodward JK, Elshaw SR, Murray AK, et al Stimulation and inhibition of uveal melanoma invasion by HGF, GRO,IL-1, and TGF Invest Ophthalmol Vis Sci 2002,43: 3144-52
[50]Cools-Lartigue J, Cesare S, Marshall JC, et al Secretion of hepatocyte growth factor and vascular endothelial growth factor during uveal melanoma-monocyte in vitro interactions. Melanoma Res 2005; 15:141-145
[51]Peruzzi B, Bottaro DP, Targeting the c-Met signaling pathway in cancer. Clin Cancer Res 2006; 12:3657-3660
[52]All-Ericsson C, Girnita L, Seregard S, Bartolazzi A, Jager MJ, Larsson O, Insulin-like growth factor-1 receptor in uveal melanoma a predictor for metastatic diseases and a potential therapeutic target. Invest Ophthalmol Vis Sci 2002; 43:1-8
[53]Riedermann J, Macaulay VM, Insulin signaling and its inhibition. Endocr Relat Cancer 2006,13 suppl 1 s33-43
[54]Girnita L, All-Ericsson C, Economou MA, et al The insulin-like growth factor-1 receptor inhibitor picropodophyllin causes tumor regression and attenuates mechanism involved in invasion of uveal melanoma cells. Clin Cancer Res 2006,12: 1383-91
[55]Bedikian AY, Legha SS, Mavligit G, et al. Treatment of uveal melanoma metastatic tothe liver:a review of the MD Anderson Cancer Center experience and prognostic factors. Cancer.1995;76:1665-1670.
[56]Bedikian AY, Kantarjian H, Young SE, et al. Prognosis in metastatic choroidal melanoma. South Med J.1981;74:574-577.
[57]Cantore M, Fiorentini G, Aitini E, et al. Intra-arterial hepatic carboplatin-based chemotherapy for ocular melanoma metastatic to the liver. Report of a phase Ⅱ study. Tumori.1994;80:37-39.
[58]Hicks C, Foss AJ, Hungerford JL. Predictive power of screening tests for metastasis in uveal melanoma. Eye.1998;12:945-948.
[59]Hahka-Kemppinen M, Muhonen T, Kangas L, et al. Chemosensitivity of human melanoma metastases in mouse subrenal capsule assay—can it predict tumour response to combined cytostatic plus interferon therapy in metastatic melanoma? Melanoma Res.1996;6:215-221.
[60]Pyrhnen S, Hahka-Kemppinen M, Muhonen T, et al. Chemoimmunotherapy with bleomycin, vincristine, lomustine, dacarbazine (BOLD), and human leukocyte interferon for metastatic uveal melanoma. Cancer.2002;95:2366-2372.
[61]Nathan FE, Berd D, Sato T, et al. BOLD+ interferon in the treatment of metastatic uveal melanoma:first report of active systemic therapy. J Exp Clin Cancer Res.1997; 16:201-208.
[62]Kivela T, Suciu S, Hansson J, et al. Bleomycin, vincristine, lomustine and dacarbazine(BOLD) in combination with recombinant interferon alpha-2b for metastatic uveal melanoma. Eur J Cancer.2003;39:1115-1120.
[63]Leyvraz S, Spataro V, Bauer J, et al. Treatment of ocular melanoma metastatic to theliver by hepatic arterial chemotherapy. J Clin Oncol.1997;15:2589-2595.
[64]Neale MH, Myatt N, Cree IA, et al. Combination chemotherapy for choroidal melanoma:ex vivo sensitivity to treosulfan with gemcitabine or cytosine arabinoside. BrJ Cancer.1999;79:1487-1493.
[65]Neale MH, Myatt NE, Khoury GG, et al. Comparison of the ex vivo chemosensitivity of uveal and cutaneous melanoma. Melanoma Res.2001;11:601-609.
[66]Terheyden P, Bro cker EB, Becker JC. Clinical evaluation of in vitro chemosensitivity testing:the example of uveal melanoma. J Cancer Res Clin Oncol. 2004;130:395-399.
[67]Pfohler C, Cree IA, Ugurel S, et al. Treosulfan and gemcitabine in metastatic uveal melanoma patients:results of a multicenter feasibility study. Anticancer Drugs. 2003;14:337-340.
[68]Keilholz U, Schuster R, Schmittel A, et al. A clinical phase I trial of gemcitabine and treosulfan in uveal melanoma and other solid tumours. Eur J Cancer.2004; 40: 2047-2052.
[69]Salmon RJ, Levy C, Plancher C, et al. Treatment of liver metastases from uveal melanoma by combined surgery-chemotherapy. Eur J Surg Oncol.1998;24:127-130.
[70]Singh AD, Borden EC. Metastatic uveal melanoma. Ophthalmol Clin North Am. 2005;18:143-150.
[71]Egerer G, Lehnert T, Max R, et al. Pilot study of hepatic intraarterial fotemustine chemotherapy for liver metastases from uveal melanoma:a single-center experience with seven patients. Int J Clin Oncol.2001;6:25-28.
[72]Becker JC, Terheyden P, Ka mpgen E, et al. Treatment of disseminated ocular melanoma with sequential fotemustine, interferon alpha, and interleukin 2. Br J Cancer.2002;87:840-845.
[73]Tojo D, Wenig BL, Resnick KI. Incidence of cervical metastasis from uveal melanoma:implications for treatment. Head Neck.1995;17:137-139.
[74]Elias D, Cavalcanti de Albuquerque A, Eggenspieler P, et al. Resection of liver metastases from a noncolorectal primary:indications and results based on 147 monocentric patients. J Am Coll Surg.1998;187:487-493.
[75]Spitler LE, Wong P, Sagebiel R. Combined immunotherapy of malignant melanoma. Unusual survival following cerebral metastasis. Arch Dermatol. 1978;114:1501-1504.
[76]Zimmerman L, McLean I. Do growth and onset of symptoms of uveal melanomas indicate subclinical metastasis? Ophthalmology 1984; 92:685-91.
[77]Donoso L, Shields J, Augsburger J et al. Metastatic uveal melanoma;Diffuse hepatic metastasis in a patient with concurrent normal serum enzyme levels and liver scan (letter). Arch Ophthalmol 1985; 103:758.
[78]Gragoudas ES, Egan KM, Seddon JM, et al. Survival of patients with metastases from uveal melanoma. Ophthalmology.1991;98:383-389.
[79]Toivonen P, Ma " kitie T, Kujala E, et al. Microcirculation and tumor-infiltrating macrophages in choroidal and ciliary body melanoma and corresponding metastases. Invest Ophthalmol Vis Sci.2004;45:1-6.
[80]Woll E, Bedikian A, Legha SS. Uveal melanoma:natural history and treatment options for metastatic disease. Melanoma Res.1999;9:575-581.
[2]Zimmerman L, McLean I. Do growth and onset of symptoms of uveal melanomas indicate subclinical metastasis? Ophthalmology 1984; 92:685-91.
[3]Donoso L, Shields J, Augsburger J et al. Metastatic uveal melanoma;Diffuse hepatic metastasis in a patient with concurrent normal serum enzyme levels and liver scan (letter). Arch Ophthalmol 1985; 103:758.
[4]Egan KM, Seddon JM, Glym RJ, et al. Edidemiologic aspects of uveal melanoma. Sur Ophthalmol,1988,32:239-251
[5]Mooy C, De Jong P. Prognostic parameters in uveal melanoma:A review. Surv Ophthalmol 1996; 41:215-28.
[6]李鹏程,张虹,丁正平,等TGF β RII和Smad4蛋白在葡萄膜黑色素瘤中的表达[J].肿瘤防治研究,2005,32(1)：35-36.
[7]吴中耀,郭琳洁,郑湖玲.葡萄膜黑色素瘤转移相关基因的表达及意义[J].中国实用眼科杂志,2001,19(2)：342-345.
[8]Al-Jamal RT, Makitie T, Kivela T. Nucleolar diameter and microvascular factors as independent predictors of mortality from malignant melanoma of the choroid and ciliary body. Invest Ophthalmol Vis Sci 2003; 44:2381-2389.
[9]郭琳洁,吴中耀,郑湖玲.细胞周期相关基因在葡萄膜黑色素瘤中表达与意义[J].中华眼底病杂志,2001,17(1)：44-46.
[10]傅涛,魏文斌,王阳.血清黑色素瘤活动抑制蛋白与葡萄膜黑色素瘤[J].中华眼底病杂志,2005,21(1)：153-155.
[11]陈陆霞,张诗武,孙保存.MMP22、MMP29的表达与脉络膜黑色素瘤微血管密度及肿瘤转移的关系[J].眼科新进展,2007,8(27)：565-567
[12]Chen JM, Dando PM, Rawlings ND, et al. Cloning, isolation, and characterization of mammalian legumain, an asparaginyl endopeptidase. Biol Chem 1997;272:8090-8.
[13]Jessica G, Frank T, Hans J. Legumain expression as a prognostic factor in breast cancer patients. Breast Cancer Res Treat 2007; 102:1-6
[14]Raghavendra V M, Gunnar A, Jingfang G. Legumain Expression in Relation to Clinicopathologic and Biological Variables in Colorectal Cancer. Clinical Cancer Research 2005; 11:2293-2299.
[15]Watts C, Matthews SP, Mazzeo D, et al. Asparaginyl endopeptidase:case history of a class ⅡMHC compartment p rotease [J]. Immunol Rev,2005,207:218-228.
[16]Luo Y, Zhou H, Krueger J, et al. Targeting tumor-associated macrophages as a novel strategy against breast cancer [J]. J Clin Invest,2006,116 (8):2132-2141.
[17]孙为荣.眼科病理学[M].北京：人民卫生出版社,1997.266-280.
[18]Zimmerman L, McLean I. Do growth and onset of symptoms of uveal melanomas indicate subclinical metastasis? Ophthalmology 1984; 92:685-91.
[19]Donoso L, Shields J, Augsburger J et al. Metastatic uveal melanoma;Diffuse hepatic metastasis in a patient with concurrent normal serum enzyme levels and liver scan (letter). Arch Ophthalmol 1985; 103:758.
[20]Chen JM, Dando PM, Rawlings ND, et al. Cloning, isolation, and characterization of mammalian legumain, an asparaginyl endopeptidase. Biol Chem 1997;272:8090-8.
[21]Watts C, Matthews SP, Mazzeo D, et al. Asparaginyl endopeptidase:case history of a class Ⅱ MHC compartment p rotease [J]. Immunol Rev,2005,207:218-228.
[22]Manoury, B., Hewitt, E. W., Morrice, N., Dando, P. M., Barrett, A. J., and Watts, C.An asparaginyl endopeptidase processes a microbial antigen for class II MHC presentation. Nature,396:695-699,1998.
[23]Choi, S. J., Kurihara, N., Oba, Y., and Roodman, G. D. Osteoclast inhibitory peptide inhibits osteoclast formation via its C-terminal fragment. J. Bone Miner. Res., 16:1804-1811,2001.
[24]Cheng L, Chengzao S, Haining H, Kim J, et al Overexpression of legumain in tumorsis significant for invasion/metastasis and a candidate enzymatic target for prodrug therapy. Cancer Res 2003; 63:2957-2964,
[25]Chen JM, Dando PM, Stevens RA. Cloning and expression of mouse legumain, a lysosmal endopeptidase. Biochem J,1988,335:111-117.
[26]Albini A, Iwamoto Y, Kleinman HK, et al. A rapid in vitro assay for quantitating the invasive potential of tumor cells. Cancer Res,1987,47(12):3239 —3245.
[27]吴文苑,余涟,刘春,罗金刚Legumain对肿瘤细胞侵润力和血管内皮细胞管腔形成的作用.热带医学杂志,2009,9(8)：864-867
[28]Susanna L, He Z, Huai-dong H, et al A Legumain-based minigene vaccine targets the tumor stromaand suppresses breast cancer growth and angiogenesis Cancer Immunol Immunother (2008) 57:507-515
[29]Luo Y, Zhou H, Krueger J, Kaplan C, Lee SH, Dolman C, Markowitz D, Wu W, Liu C, Reisfeld RA, Xiang R. Targeting tumor-associated macrophages as a novel strategy against breast cancer. J Clin Invest 2006; 116:2132
[30]Jedeszko C, Sloane BF. Cysteine cathepsins in human cancer. Biol Chem,2004, 385 (11):1017-1027.
[31]Yoshikata M, Hisazumi A, Toshiro S, et al. Legumain/asparaginyl endopeptidase controls extracellular matrix remodeling through the degradation of fibronectin in mouse renal proximal tubular cells FEBS Lett.2007,581 (7):1417-1424.
[32]Bingle L, Brown NJ, Lewis CE. The role of tumor-associated macrophages in tumor progression:imp lications for new anticancer therapies[J]. J Pathol,2002, 196 (3):254-256.
[33]Lewis CE, Pollard JW. Distinct role of macrophages in different tumor micro-environments[J]. Cancer Res,2006,66 (2):605-612.
[34]郭强,李康,徐薇,储以微,熊思东.Legumain在荷瘤与正常小鼠脾细胞中的差异性表达.中国癌症杂志,2008,18(2)：101-105
[35]Wolk K, Grutz G, Witte K, et al. The exp ression of Legumain, an asparaginyl endopep tidase that controls antigen p rocessing, is reduced in endotoxin-tolerant monocytes [J]. Genes Immun,2005,6 (5):452-456.
[36]Luo Y, Zhou H, Liu C, et al. A novel stress protein, legumain, is a target for a genetic vaccine against breast cancer [A]. AACR Meeting Abstracts [C],2005,5 (1):1216-1217.
[37]Elisabeth AS, Paul SM, Dawn AK, Jacob P, Andrew JM, Jeffrey MT, Robert F& Mary J.C. Hendrix. Molecular determinants of human uveal melanoma invasion and metastasis. Clinical & Experimental Metastasis 19:233-246,2002.
[38]颜建华,吴中耀.葡萄膜肿瘤.见：吴中耀,主编.现代眼肿瘤眼眶病学.北京：人民军医出版社,2002.222-232.
[39]王风华,李彬.脉络膜黑色素瘤的临床、组织病理学特点及研究进展.国外医学眼科学分册,2001,25：372-377.
[40]McLean IW, Foster WD, Zimmerman L E, et al. Modifications of Callender's classification of uveal melanoma at the Armed Forces Institute of Pathology. Am J Ophthalmol,1983,96:502-509.
[41]梁建宏,易长贤,Schilling H,等.脉络膜恶性黑色素瘤中Ki267的表达和肿瘤主要组织病理特性的关系.眼科学报,2001,17：114-117.
[42]林锦镛.葡萄膜黑色素瘤的临床病理学特征和鉴别诊断要点.中华眼科杂志,2003,39：121-124.
[43]罗清礼,孙宪丽.努力提高脉络膜黑色素瘤的基础和临床研究水平.中华眼科杂志,2003,39：65267.
[44]Coleman K, Baak J A, Van Diest P, Mullaney J, Farrell M & Fenton M Prognostic factors following enucleation of 111 uveal melanomas. Br J Ophthalmol 1993,77:688-692
[45]Shields J A (1992):Posterior uveal melanoma:clinical and pathological features. In:Shields J A & Shields C L (eds). Intraocular tumors. A text and atlas. WB Saunders Co, Philadelphia.
[46]Kujala E, Makitie T, Kivela T. Very long-term prognosis of patients with malignant uveal melanoma. Invest Ophthalmol Vis Sci 2003;44:4651-9.
[47]Scholes AG, Damato BE, Nunn J, Hiscott P, Grierson I & Field JK:Monosomy 3 in uveal melanoma:correlation with clinical and histologic predictors of survival. Invest Ophthalmol Vis Sci 2003,44:1008-1011.
[48]Makitie T, Summanen P, Tarkkanen A &Kivela T:Microvascular density in predicting survival of patients with choroidal and ciliary body melanoma. Invest Ophthalmol Vis Sci 1999,40:2471-2480.
[49]Seregard S, Kock E. Prognostic indicators following enucleation for posterior uveal melanoma:a multivariate analysis of long-term survival with minimized loss to follow-up. Acta Ophthalmol Scand 1995;73:340-4.
[50]郭琳洁,吴中耀,张胜,陈家祺,艾思明,郑湖玲.葡萄膜黑色素瘤病理学 特征与预后的关系.中华病理学杂志,2002,31(6)：518-522
[51]Sarah E, Ian C, Bertil D. Routes of Extraocular Extension of Uveal Melanoma Ophthalmology.2008; 115 (10):1778-1785
[52]Patel KA, Edmondson ND, Talbot F,Parsons MA, Rennie IG & Sisley K Prediction of prognosis in patients with uveal melanoma using fluorescence in situ hybridization. Br J Ophthalmol 2001,85:1440-1444.
[53]Haddab S, Hidayat A, Tabbara KF. Ciliary body melanoma with optic nerve invasion. Br J Ophthalmol.1990;74:123-124.
[54]Chess J, Albert DM, Bellows AR, Dallow R. Uveal melanoma:case report of extension through the optic nerve to the surgical margin in the orbital apex. Br J Ophthalmol.1984;68:272-275.
[55]Abdel-Rahman MH, Craig EL, Davidorf FH, Eng C. Expression of vascular endothelial growth factor in uveal melanoma is independent of 6p21-region copy number. Clin Cancer Res.2005; 11:73-78.
[56]Spencer WH. Optic nerve extension of intraocular neoplasms. Am J Ophthalmol. 1975;80:465-471.
[57]Walker TM, Van Ginkel PR, Gee RL, et al. Expression of angiogenic factors Cyr61 and tissue factor in uveal melanoma. Arch Ophthalmol.2002;120:1719-1725.
[58]Boyd SR, Tan DS, de Souza L, et al. Uveal melanomas express vascular endothelial growth factor and basic fibroblast growth factor and support endothelial cell growth. Br J Ophthalmol.2002;86:440-447.
[59]Cohn-Cedermark G, Mansson-Brahme E, Rutqvist LE, Larsson O, Johansson H, Ringborg U. Central nervous system metastases of cutaneous malignant melanoma—a population-based study. Acta Oncol.1998;37:463-470.
[60]Donoso LA, Nagy RM, Brockman RJ, et al. Metastatic uveal melanoma. Hepatic cell surface enzymes, isoenzymes, and serum sialic acid levels in early metastatic disease.Arch Ophthalmol.1983;101:791-794.
[61]Schaller UC, Bosserhoff AK, Neubauer AS, et al. Melanoma inhibitory activity: a novel serum marker for uveal melanoma. Melanoma Res.2002; 12:593-599.
[62]Maniotis AJ, Folberg R, Hess A, et al. Vascular channel formation by human melanoma cells in vivo and in vitro:vasculogenic mimicry. Am J Pathol,1999, 155:739-752
[63]Gawenda J, Traub F, Luck HJ, et al. Legumain expression as a prognostic factor in breast cancer patients. Breast Cancer Res Treat.2007,102 (1):126.
[64]Murthy RV, Arbman G, Gao J, Roodman GD, Sun XF. Legumain expression in relation to clinicopathologic and biological variables in colorectal cancer. Clin Cancer Res 2005; 11:2293
[65]Beck, H., Schwarz, G., Schroter, C. J., Deeg, M., Baier, D., Stevanovic, S., Weber,E., Driessen, C., and Kalbacher, H. Cathepsin S and an asparagine-specific endoprotease dominate the proteolytic processing of human myelin basic protein in vitro. Eur. J. Immunol.,2001.31:3726-3736,
[66]Wu W, Luo Y, Sun C, et al. Targeting cell-impermeable prodrug activation to tumor microenvironment eradicates multiple drug-resistant neoplasms [J]. Cancer Res,2006,66 (2):970-980.
[1]Sobin LH, Wittekind Ch. TNM Classification of Malignant Tumours. New York: Wiley-Liss; 2002.
[2]Lorigan JG, Wallace S, Mavligit GM. The prevalence and location of metastases from ocular melanoma:imaging study in 110 patients. AJR Am J Roentgenol. 1991;157:1279-
[3]Aoyama T, Mastrangelo MJ, Berd D, et al. Protracted survival after resection of metastatic uveal melanoma. Cancer.2000;89:1561-1568.
[4]Eskelin S, Pyrhonen S, Summanen P, et al. Screening for metastatic malignant melanoma of the uvea revisited. Cancer.1999;85:1151-1159.
[5]Bergman L, Seregard S, Nilsson B, et al. Uveal melanoma survival in Sweden from 1960 to 1998. Invest Ophthalmol Vis Sci.2003;44:3282-3287.
[6]Kujala E, Ma T, Kivela T. Very long-term prognosis of patients with malignant uveal melanoma. Invest Ophthalmol Vis Sci.2003;44:4651-4659.1281.
[7]Collins VP, Loeffler RK, Tivey H. Observations on growth rates of human tumors. AJR Am J Roentgenol.1956;76:988-1000.
[8]Eskelin S, Pyrho S, Summanen P, et al. Tumor doubling times in metastatic malignant melanoma of the uvea:tumor progression before and after treatment. Ophthalmology.2000; 107:1443-1449.
[9]Singh AD. Uveal melanoma:implications of tumor doubling time [Letter]. Ophthalmology.2001;108:829-831.
[10]Manschot WA, van Peperzeel HA. Choroidal melanoma. Enucleation or observation? A new approach. Arch Ophthalmol.1980;98:71-77.
[11]Patel JK, Didolkar MS, Pickren JW, et al. Metastatic pattern of malignant melanoma. A study of 216 autopsy cases. Am J Surg.1978;135:807-810.
[12]Gombos DS, Van Quill KR, Uusitalo M, et al. Geographic disparities in diagnostic screening for metastatic uveal melanoma. Ophthalmology.2004; 111:2254-2258.
[13]Diener-West M, Reynolds SM, Agugliaro DJ, et al. Screening for metastasis from choroidal melanoma:the Collaborative Ocular Melanoma Study Group Report 23. J Clin Oncol.2004;22:2438-2444.
[14]Eskelin S, Pyrho nen S, Hahka-Kemppinen M, et al. A prognostic model and staging for metastatic uveal melanoma. Cancer.2003;97:465-475.
[15]Kaiserman I, Amer R. Pe'er J. Liver function tests in metastatic uveal melanoma. Am J Ophthalmol.2004;137:236-243.
[16]Eskelin S, Pyrhonen S, Summanen P, et al. Screening for metastatic malignant melanoma of the uvea revisited. Cancer.1999;85:1151-1159.
[17]The Collaborative Ocular Melanoma Study Group. Assessment of metastatic diseasestatus at death in 435 patients with large choroidal melanoma in the Collaborative Ocular Melanoma Study (COMS):COMS report no.15. Arch Ophthalmol.2001;119:670-676.
[18]The Collaborative Ocular Melanoma Study Group. The Collaborative Ocular Melanoma Study (COMS) randomized trial of pre-enucleation radiation of large choroidal melanoma Ⅰ:characteristics of patients enrolled and not enrolled. COMS report no.9. Am J Ophthalmol.1998;125:767-778.
[19]Missotten GS, Tang NE, Korse CM, et al. Prognostic value of S-100-beta serum concentration in patients with uveal melanoma. Arch Ophthalmol.2003; 121:1117 1119.
[20]Schaller UC, Bosserhoff AK, Neubauer AS, et al. Melanoma inhibitory activity: a novel serum marker for uveal melanoma. Melanoma Res.2002;12:593-599.
[21]Donoso LA, Berd D, Augsburger JJ, et al. Metastatic uveal melanoma. Pretherapy serum liver enzyme and liver scan abnormalities. Arch Ophthalmol. 1985;103:796-798.
[22]Donoso LA, Nagy RM, Brockman RJ, et al. Metastatic uveal melanoma. Hepatic cellsurface enzymes, isoenzymes, and serum sialic acid levels in early metastatic disease. Arch Ophthalmol.1983;101:791-794.
[23]Diener-West M, Earle JD, Fine SL, et al. The COMS randomized trial of iodine
125 brachytherapy for choroidal melanoma, Ⅱ:characteristics of patients enrolled and not enrolled. COMS report no.17. Arch Ophthalmol.2001; 119:951-965.
[24]Myatt N, Aristodemou P, Neale MH, et al. Abnormalities of the transforming growth factor-beta pathway in ocular melanoma [J]. J Pathol,2000,192:511-518
[25]Sun L. Tumor-supp ressive and p romoting function of transforming growth factor beta [J]. Front Biosci,2004,9:1925-1935
[26]Ramont L, Pasco S, Hornebeck W, et al. Transforming growth factor-betal inhibits tumor growth in a mouse melanoma model by down-regulating the plasminogen activation system[J]. Exp Cell Res,2003,291: 1-10
[27]Park SS,Li L, Korn TS, et al. Effect of transforming growth factor2beta on plasminogen activator p roduction of cultured human uvealmelanoma cells [J]. Curr Eye Res,1996,15:755-763
[28]Ijland SA, Jager MJ, Heijdra BM, et al. Exp ression of angiogenic and immunosuppressive factors by uveal melanoma cell lines [J]. Melanoma Res,1999, 9:445-450
[29]EI-Shabrawi Y, Ardjomand N, Radner H, et al MMP-9 is predominantly expressed in epithelioid and not spindle cell uveal melanoma. J Pathol 2001; 194 (3): 201-206
[30]Foda H, Zucker S. Matrix metallop roteinases in cancer invasion metastasis and angiogenesis. Drug Discov Today 2001; 6:478-482
[31]Garbell EA, ReedMW, Brown NJ, et a.l Proleolysis in human breast and colorectal cancer. Br J Cancer 1999; 81 (2):287-293
[32]Gilles C, PoletteM, Coraux C. Contribution ofMT12MMP and of human laminin25 gamma22 chain degradation to mammary epithelial cell migration. J Cell Scil 2001; 11 (4):2967-2976
[33]Hotary K, Allen E, Puntrieri A. Regulation of cell invasion and morphogenesis in a three-dimensional type I collagen matrix by membrane type metallop roteinase 1,2 and 3. J Cell Biol 2000; 149 (6):1309-1323
[34]Baker JK, Elshaw SR, Mathewman GE, et a.l Expression of integrins, degradeative enzymes and their inhibitors in uveal melanoma:differences between invitro and invivo expression. Melan Res 2001; 11(3):265-273
[35]Broooks PC, Silllettl S, Von Schalscha TL. Disruption of angiogenesis by PEX, a noncatalytic metalloproteinase fragment thintegring binding cativity. Cell 1998; 92 (3):391-400
[36]Fang J, Shing Y, Wiederschaim D, et a.l Matrix Matallop roteinase-2 is required for the switch to the angiogenic phenol-type in a tumor model. ProcNatlA cad Sci USA 2000; 1997 (8):3884-3889
[37]Vannen A, Kallioinen M, von Dickh off K, et a.l. Matrix metallop roteinase-2 (MMP-2) immunoreactive protein a new prognostic marker in uveal melanoma. J Pathol 1999; 188 (1):56-62
[38]陈陆霞,张诗武,孙保存,等.MMP22、MMP29的表达与脉络膜黑色素瘤微血管密度及肿瘤转移的关系.眼科新进展2007；27(8)：565-567
[39]栗映梅,申家泉.基质金属蛋白酶及其组织抑制因子与眼病的相关性研究进展.国际眼科杂志2007；7(4)：1135-1138
[40]陈翔,田立红,谢红付,等.CD147对成纤维细胞MMP29的产生和恶性黑色素瘤细胞浸润的作用.中南大学学报(医学版)2005；30(3)：249-252
[41]Maniotis AJ, Folberg R, Hess A, et al. Vascular channel formation by human melanoma cells in vivo and in vit ro:vasculogenic mimicry. Am J Pathol,1999, 155:739-752
[42]Folberg R, Pe'er J, Gruman LM, et al. The morphologic characteristics of tumor blood vessels as a marker of tumor progression in primary human uveal melanoma:a matched case2control study. Hum Pathol,1992,23:1298-1305
[43]Hendrix MJ, Seftor EA, Meltzer PS, et al. Expression and functional significance of VE-cadherin in aggressive human melanoma cells:role in vasculogenic mimicry. Proc Natl Acad Sci USA,2001,98:8018-8023
[44]Bittner M, Meltzer P, Chen Y, et al. Molecular classification of cutaneous malignant melanoma by gene expression profiling. Nature,2000,406:536-540
[45]Hendrix MJ, Seftor EA, Seftor RE, et al Regulation of uveal melanoma intercoverted phenotype by hepatocyte growth factor(HGF). Am J Pathol 1998; 152: 855-863
[46]Hendrix MJ, Seftor EA, Seftor RE, et al Biologic determinants of uveal melanoma metastatic phenotype role of intermediate filaments as predictive markers. Lab Invest 1998; 78:153-163
[47]Malikarjuna K, Pushpara V, Biswas J, et al Expression of epidermal growth factor receptor, ezin, hepatocyte growth factor and c-Met in uveal melanoma an immunohistochemical study. Curr Eye Res 2007; 32:281-90
[48]Cesare S, Marshall JC, Logan P et al Expression and migratory analysis of 5 human uveal melanoma cell lines for CXCL12, CXCL8, CXCL1, and HGF. J Carcinog 2007,6:34-42
[49]Woodward JK, Elshaw SR, Murray AK, et al Stimulation and inhibition of uveal melanoma invasion by HGF, GRO,IL-1, and TGF Invest Ophthalmol Vis Sci 2002,43: 3144-52
[50]Cools-Lartigue J, Cesare S, Marshall JC, et al Secretion of hepatocyte growth factor and vascular endothelial growth factor during uveal melanoma-monocyte in vitro interactions. Melanoma Res 2005; 15:141-145
[51]Peruzzi B, Bottaro DP, Targeting the c-Met signaling pathway in cancer. Clin Cancer Res 2006; 12:3657-3660
[52]All-Ericsson C, Girnita L, Seregard S, Bartolazzi A, Jager MJ, Larsson O, Insulin-like growth factor-1 receptor in uveal melanoma a predictor for metastatic diseases and a potential therapeutic target. Invest Ophthalmol Vis Sci 2002; 43:1-8
[53]Riedermann J, Macaulay VM, Insulin signaling and its inhibition. Endocr Relat Cancer 2006,13 suppl 1 s33-43
[54]Girnita L, All-Ericsson C, Economou MA, et al The insulin-like growth factor-1 receptor inhibitor picropodophyllin causes tumor regression and attenuates mechanism involved in invasion of uveal melanoma cells. Clin Cancer Res 2006,12: 1383-91
[55]Bedikian AY, Legha SS, Mavligit G, et al. Treatment of uveal melanoma metastatic tothe liver:a review of the MD Anderson Cancer Center experience and prognostic factors. Cancer.1995;76:1665-1670.
[56]Bedikian AY, Kantarjian H, Young SE, et al. Prognosis in metastatic choroidal melanoma. South Med J.1981;74:574-577.
[57]Cantore M, Fiorentini G, Aitini E, et al. Intra-arterial hepatic carboplatin-based chemotherapy for ocular melanoma metastatic to the liver. Report of a phase Ⅱ study. Tumori.1994;80:37-39.
[58]Hicks C, Foss AJ, Hungerford JL. Predictive power of screening tests for metastasis in uveal melanoma. Eye.1998;12:945-948.
[59]Hahka-Kemppinen M, Muhonen T, Kangas L, et al. Chemosensitivity of human melanoma metastases in mouse subrenal capsule assay—can it predict tumour response to combined cytostatic plus interferon therapy in metastatic melanoma? Melanoma Res.1996;6:215-221.
[60]Pyrhnen S, Hahka-Kemppinen M, Muhonen T, et al. Chemoimmunotherapy with bleomycin, vincristine, lomustine, dacarbazine (BOLD), and human leukocyte interferon for metastatic uveal melanoma. Cancer.2002;95:2366-2372.
[61]Nathan FE, Berd D, Sato T, et al. BOLD+ interferon in the treatment of metastatic uveal melanoma:first report of active systemic therapy. J Exp Clin Cancer Res.1997; 16:201-208.
[62]Kivela T, Suciu S, Hansson J, et al. Bleomycin, vincristine, lomustine and dacarbazine(BOLD) in combination with recombinant interferon alpha-2b for metastatic uveal melanoma. Eur J Cancer.2003;39:1115-1120.
[63]Leyvraz S, Spataro V, Bauer J, et al. Treatment of ocular melanoma metastatic to theliver by hepatic arterial chemotherapy. J Clin Oncol.1997;15:2589-2595.
[64]Neale MH, Myatt N, Cree IA, et al. Combination chemotherapy for choroidal melanoma:ex vivo sensitivity to treosulfan with gemcitabine or cytosine arabinoside. BrJ Cancer.1999;79:1487-1493.
[65]Neale MH, Myatt NE, Khoury GG, et al. Comparison of the ex vivo chemosensitivity of uveal and cutaneous melanoma. Melanoma Res.2001;11:601-609.
[66]Terheyden P, Bro cker EB, Becker JC. Clinical evaluation of in vitro chemosensitivity testing:the example of uveal melanoma. J Cancer Res Clin Oncol. 2004;130:395-399.
[67]Pfohler C, Cree IA, Ugurel S, et al. Treosulfan and gemcitabine in metastatic uveal melanoma patients:results of a multicenter feasibility study. Anticancer Drugs. 2003;14:337-340.
[68]Keilholz U, Schuster R, Schmittel A, et al. A clinical phase I trial of gemcitabine and treosulfan in uveal melanoma and other solid tumours. Eur J Cancer.2004; 40: 2047-2052.
[69]Salmon RJ, Levy C, Plancher C, et al. Treatment of liver metastases from uveal melanoma by combined surgery-chemotherapy. Eur J Surg Oncol.1998;24:127-130.
[70]Singh AD, Borden EC. Metastatic uveal melanoma. Ophthalmol Clin North Am. 2005;18:143-150.
[71]Egerer G, Lehnert T, Max R, et al. Pilot study of hepatic intraarterial fotemustine chemotherapy for liver metastases from uveal melanoma:a single-center experience with seven patients. Int J Clin Oncol.2001;6:25-28.
[72]Becker JC, Terheyden P, Ka mpgen E, et al. Treatment of disseminated ocular melanoma with sequential fotemustine, interferon alpha, and interleukin 2. Br J Cancer.2002;87:840-845.
[73]Tojo D, Wenig BL, Resnick KI. Incidence of cervical metastasis from uveal melanoma:implications for treatment. Head Neck.1995;17:137-139.
[74]Elias D, Cavalcanti de Albuquerque A, Eggenspieler P, et al. Resection of liver metastases from a noncolorectal primary:indications and results based on 147 monocentric patients. J Am Coll Surg.1998;187:487-493.
[75]Spitler LE, Wong P, Sagebiel R. Combined immunotherapy of malignant melanoma. Unusual survival following cerebral metastasis. Arch Dermatol. 1978;114:1501-1504.
[76]Zimmerman L, McLean I. Do growth and onset of symptoms of uveal melanomas indicate subclinical metastasis? Ophthalmology 1984; 92:685-91.
[77]Donoso L, Shields J, Augsburger J et al. Metastatic uveal melanoma;Diffuse hepatic metastasis in a patient with concurrent normal serum enzyme levels and liver scan (letter). Arch Ophthalmol 1985; 103:758.
[78]Gragoudas ES, Egan KM, Seddon JM, et al. Survival of patients with metastases from uveal melanoma. Ophthalmology.1991;98:383-389.
[79]Toivonen P, Ma " kitie T, Kujala E, et al. Microcirculation and tumor-infiltrating macrophages in choroidal and ciliary body melanoma and corresponding metastases. Invest Ophthalmol Vis Sci.2004;45:1-6.
[80]Woll E, Bedikian A, Legha SS. Uveal melanoma:natural history and treatment options for metastatic disease. Melanoma Res.1999;9:575-581.