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      Annexin A2及CD105在胰腺導(dǎo)管腺癌中的表達(dá)及臨床意義

      2014-04-17 20:39:13黃亞楷劉宏劉金彪王新征
      中國實(shí)用醫(yī)藥 2014年6期

      黃亞楷 劉宏 劉金彪 王新征

      【摘要】 目的 探討Annexin A2 (ANXA2, 膜聯(lián)蛋白A2)及CD105在胰腺導(dǎo)管腺癌中表達(dá)及兩者的相關(guān)性。方法 采用免疫組織化學(xué)染色檢測胰腺導(dǎo)管腺癌及癌旁組織中Annexin A2的表達(dá)情況及MVD, 并對結(jié)果進(jìn)行相關(guān)性分析。結(jié)果 Annexin A2在腫瘤組織中表達(dá)的陽性率顯著高于癌旁組織(P<0.05), 且癌組織中Annexin A2的表達(dá)強(qiáng)度高于癌旁組織(P<0.05)。低分化癌組織Annexin A2表達(dá)的陽性率高于高分化和中分化癌組織的陽性率。Annexin A2的表達(dá)與癌組織的淋巴轉(zhuǎn)移相關(guān)。胰腺導(dǎo)管腺癌組織中的MVD明顯高于癌旁組織(P<0.05)。同時, MVD隨癌組織分化程度的增高而降低。TNM分期的Ⅲ~Ⅳ期明顯高于Ⅰ~Ⅱ期(P<0.05)。CD105標(biāo)記的MVD與癌組織的淋巴轉(zhuǎn)移相關(guān)。Annexin A2染色3+的癌組織MVD明顯低于Annexin A2染色0、1+、2+(P<0.05), 但Annexin A2染色0、1+、2+的癌組織的MVD表達(dá)差異無統(tǒng)計學(xué)意義(P=0.615、0.243、0.459)。結(jié)論 Annexin A2與CD105在胰腺導(dǎo)管腺癌組織中表達(dá)升高, 且都與癌組織病理分級及淋巴轉(zhuǎn)移相關(guān)。兩者具有一定相關(guān)性。

      【關(guān)鍵詞】 胰腺導(dǎo)管腺癌;Annexin A2;CD105

      Expression of Annexin A2 and CD105 in pancreatic ductal adenocarcinoma and their clinical significance HUANG Ya-kai, LIU Hong, LIIU Jin-biao, et al. Depatrment of Surgery, First Hospital Affiliated to Henan University of Science and Technology, Luoyang 471000, China

      【Abstract】 Objective To explore the expression of Annexin A2 and CD105 in pancreatic ductal adenocarcinoma(PDAC) tissues and peficancerous tissues and their clinical significance. Methords Using immunohistochemical staining to detect the expression of Annexin A2 and CD105 in pancreatic cancerous tissues and peficancerous tissues, and conduct correlation analysis of results. Results The positive expression rate of Annexin A2 and the MVD in pancreatic ductal adenocarcinoma tissues was higher than that it in peficancerous tissues(P<0.05), the express degree of Annexin A2 in cancerous tissues was higher than it in peficancerous tissues(P<0.05). The expression of Annexin A2 and the MVD were correlated with histological grade and lymphatic metastasis(P<0.05). The MVD of cancerous tissues in clinical stage Ⅲ~Ⅳ was significantly higher than that in clinical stageⅠ~Ⅱ(P<0.05). The MVD of Annexin A2 dyeing 3 + cancerous tissue was significantly lower than those in other Annexin A2 dyeing grade(P<0.05). Conclusion The expression of Annexin A2 and CD105 in PDAC is higher than it in peficancerous tissues, and two of both are correlated with hitological grade and lymphatic metastasis. There is correlation in both.

      【Key words】 Pancreatic ductal adenocarcinoma; Annexin A2; CD105

      Annexin A2 ( ANXA2) 又名膜聯(lián)蛋白Ⅱ, 屬于膜聯(lián)蛋白家族的成員, 與多種腫瘤的發(fā)生發(fā)展密切相關(guān), Annexin A2的表達(dá)失調(diào)與腫瘤的侵襲、轉(zhuǎn)移密切相關(guān)[1-3]。CD105作為一種細(xì)胞膜表面的糖蛋白[4], 是血管內(nèi)皮細(xì)胞增殖的標(biāo)志之一[5], CD105所標(biāo)記的MVD可更好的反映癌組織中的新生血管數(shù)量。有關(guān)聯(lián)合檢測Annexin A2、CD105蛋白在胰腺導(dǎo)管腺癌的表達(dá)及與胰腺導(dǎo)管腺癌臨床及病理學(xué)參數(shù)關(guān)系之間的研究, 國內(nèi)尚未見文獻(xiàn)報道, 為進(jìn)一步探討兩者與胰腺導(dǎo)管腺癌臨床及病理學(xué)參數(shù)及兩者之間的相互聯(lián)系, 本實(shí)驗(yàn)采用免疫組織化學(xué)Power Vision 法檢測Annexin A2、CD105蛋白在胰腺導(dǎo)管腺癌組織及癌旁組織中的表達(dá), 探討兩者與胰腺導(dǎo)管腺癌臨床及病理學(xué)參數(shù)之間的關(guān)系及可能的機(jī)制。

      1 資料與方法

      1. 1 標(biāo)本來源 于山西醫(yī)科大學(xué)第一醫(yī)院病理科選擇51例胰腺癌組織標(biāo)本及相應(yīng)胰腺組織陰性切緣標(biāo)本, 均使用10%中性福爾馬林固定并石蠟包埋。所有病例均來源于2005~2010年, 其中男性24例, 女性27例, 年齡31~78歲, 中位年齡59.15歲。51例患者胰腺癌組織標(biāo)本均經(jīng)病理學(xué)診斷均為胰腺導(dǎo)管腺癌, 其中高分化腺癌(G1級)14例、中分化腺癌(G2級)6例、低分化腺癌(G3級)31例, 胰腺癌的TNM分期參照UICC2002年標(biāo)準(zhǔn), Ⅰ~Ⅳ期分別為16例、7例、20例、8例。所有患者術(shù)前均未進(jìn)行過放療及化療。

      1. 2 實(shí)驗(yàn)試劑 兔抗人 Annexin A2多克隆抗體購自武漢博士德生物工程有限公司, 鼠抗人CD105單克隆抗體、兔抗及鼠抗免疫組化二步法試劑盒和DAB顯色試劑盒均購自北京中杉金橋生物技術(shù)有限公司。

      1. 3 實(shí)驗(yàn)方法 Annexin A2、CD105蛋白檢測采用免疫組化Power Vision 法, 操作按試劑盒說明書進(jìn)行。標(biāo)本常規(guī)石蠟包埋, 4 μm切片, 進(jìn)行免疫組化染色。用已知陽性切片做陽性對照, PBS代替一抗為陰性對照。DAB染色后蘇木素對比復(fù)染, 中性樹膠封片。

      1. 4 結(jié)果判定 Annexin A2蛋白表達(dá)情況結(jié)果判定標(biāo)準(zhǔn)為:胞漿或胞膜出現(xiàn)淡黃至棕黃或棕褐色顆粒為陽性細(xì)胞。按染色強(qiáng)度及陽性細(xì)胞數(shù)來計算評分:①染色強(qiáng)度:0分為陰性, 1分為淡黃色, 2分為黃色, 3分為棕黃色。②陽性細(xì)胞數(shù):0分為無陽性細(xì)胞, 1分為陽性細(xì)胞數(shù)≤25%, 2分為陽性細(xì)胞數(shù)26%~50%, 3分為陽性細(xì)胞數(shù)≥51%。①+②之和即為最后得分, 0分為“0”, 1~2分為“1+”, 3~4分為“2+”, 5~6分為“3+”, “+”~“3+”判定為免疫組織化學(xué)染色陽性;CD105標(biāo)志的微血管密度 (microvessel density, MVD)計數(shù)方法:先在低倍鏡下(40X)選取微血管數(shù)量最豐富的3個區(qū)域, 然后在400倍視野下分別計數(shù)上述3個區(qū)域的微血管數(shù), 取其平均值作為MVD。若發(fā)現(xiàn)和鄰近的微血管、腫瘤細(xì)胞或其他結(jié)締組織分開的單個黃褐色內(nèi)皮細(xì)胞或黃褐色內(nèi)皮細(xì)胞簇, 就計數(shù)為一個獨(dú)立的微血管, 管腔>8個紅細(xì)胞直徑或有管壁平滑肌的大血管不作為新生血管計數(shù)。

      1. 5 統(tǒng)計學(xué)方法 采用SPSS13.0統(tǒng)計軟件進(jìn)行統(tǒng)計分析, Annexin A2的表達(dá)情況及相關(guān)性分析采用Chi-square檢驗(yàn)、秩和檢驗(yàn)及Fisher精確概率法;MVD以均數(shù)±標(biāo)準(zhǔn)差表示, 其相關(guān)性分析采用t檢驗(yàn)和方差分析, 且在統(tǒng)計分析前行正態(tài)性檢驗(yàn)和方差齊性檢驗(yàn);以α=0.05為檢驗(yàn)水準(zhǔn)。

      2 結(jié)果

      2. 1 Annexin A2的表達(dá) Annexin A2在腫瘤組織細(xì)胞膜上的表達(dá)明顯高于癌旁組織, 腫瘤組織的細(xì)胞膜呈棕黃色(見圖1);而癌旁胰腺組織的細(xì)胞膜多不著色(見圖2)。腫瘤組織中Annexin A2表達(dá)的陽性率為88.2%(45/51), 顯著高于癌旁組織(33.3%, 17/51)(P<0.05)(見表1), 且腫瘤組織中Annexin A2表達(dá)的陽性強(qiáng)度高于癌旁組織(P<0.05) (見表2)。Annexin A2在低分化腫瘤組織中表達(dá)的陽性率為96.8%(30/31), 高于高分化和中分化腫瘤組織(75%, 15/20)(見表3)。在有淋巴結(jié)轉(zhuǎn)移的腫瘤組織中Annexin A2表達(dá)的陽性率為96.55%(28/29), 高于無淋巴結(jié)轉(zhuǎn)移組的77.27%(P=0.047, 17/22)(見表3), Annexin A2 的表達(dá)與患者性別、年齡、原發(fā)腫瘤的大小及臨床分期無關(guān)(見表3)。

      2. 2 CD105的表達(dá) CD105主要表達(dá)于新生血管內(nèi)皮細(xì)胞的胞膜和胞質(zhì)中, 呈棕黃色顆粒狀, 這些新生血管可不形成明顯的管腔(見圖3)。在腫瘤組織中的非新生血管內(nèi)皮細(xì)胞及在癌旁組織中, CD105弱表達(dá)或陰性表達(dá)(見圖4)。腫瘤組織中MVD均值為(10.54±3.35), 顯著高于癌旁組織的(1.08±1.163)(P<0.05)(見表1)。不同分化程度的胰腺導(dǎo)管腺癌組織的MVD不同, 低分化最高, 高分化最低(見表3);TNM分期Ⅲ~Ⅳ期明顯高于Ⅰ~Ⅱ期(P<0.01)(見表3);有淋巴結(jié)轉(zhuǎn)移組顯著高于無淋巴結(jié)轉(zhuǎn)移組(P<0.01)(見表3);CD105標(biāo)記的MVD與患者的性別、年齡、原發(fā)腫瘤的大小無關(guān)(見表3)。

      2. 3 胰腺導(dǎo)管腺癌組織中Annexin A2表達(dá)與MVD的相關(guān)性:Annexin A2染色3+腫瘤組織的MVD明顯低于Annexin A2染色0、1+、2+腫瘤組織(P<0.05), 但Annexin A2染色0、1+、2+的腫瘤組織之間的MVD差異無統(tǒng)計學(xué)意義(見表4、表5)。

      3 討論

      Annexin A2屬Annexin(膜聯(lián)蛋白)家族成員, 廣泛存在于人體內(nèi)多種細(xì)胞內(nèi)[6], 通過調(diào)控胞內(nèi)鈣的釋放而調(diào)節(jié)細(xì)胞的功能[7]。研究顯示Annexin A2在腫瘤的發(fā)生中起著重要作用[8]。Annexin A2的異四聚體(AIIt)可作為t-PA 和PLG的共受體, 調(diào)節(jié)Plasmin 的產(chǎn)生, 進(jìn)而促進(jìn)腫瘤的浸潤和轉(zhuǎn)移[9]。文獻(xiàn)報道多組蛋白質(zhì)組學(xué)研究顯示Annexin A2為胰腺癌表達(dá)的差異蛋白, 本實(shí)驗(yàn)也發(fā)現(xiàn)Annexin A2在胰腺癌組織中高表達(dá), 更進(jìn)一步證實(shí)Annexin A2有可能作為胰腺癌的分子標(biāo)志物。本實(shí)驗(yàn)免疫組化結(jié)果顯示Annexin A2在胰腺導(dǎo)管腺癌組織中表達(dá)增高且與腫瘤組織病理分級相關(guān)。因此, Annexin A2可能成為一個潛在的胰腺導(dǎo)管腺癌診斷標(biāo)志物。同時推測, Annexin A2在腫瘤侵襲方面的重要作用也使其有可能成為治療胰腺導(dǎo)管腺癌的靶點(diǎn)。

      CD105 (Endoglin)屬于轉(zhuǎn)化生長因子β(TGFβ)超家族成員, 與血管發(fā)生及維持血管完整性有關(guān), 參與腫瘤血管生成。研究表明, CD105 在新生血管內(nèi)皮細(xì)胞上強(qiáng)表達(dá), 是血管內(nèi)皮細(xì)胞增殖的標(biāo)志之一[10], CD105在腫瘤組織周圍的動、靜脈內(nèi)皮細(xì)胞中強(qiáng)表達(dá), 少數(shù)情況下也可在腫瘤細(xì)胞中弱表達(dá)[11], 本實(shí)驗(yàn)證實(shí)CD105主要表達(dá)于腫瘤組織邊緣的血管內(nèi)皮細(xì)胞中。腫瘤組織中的微血管密度(MVD)與癌癥患者的預(yù)后直接相關(guān), 文獻(xiàn)顯示在乳腺癌、宮頸癌等惡性腫瘤組織中, CD105標(biāo)記MVD值優(yōu)于CD34、Ⅷ因子相關(guān)抗原等的標(biāo)記結(jié)果, 并與患者的生存期有關(guān), 可以作為獨(dú)立的預(yù)后因子[12]。本實(shí)驗(yàn)證實(shí)胰腺導(dǎo)管腺癌組織中CD105標(biāo)記的MVD明顯高于癌旁組織, 且與腫瘤組織的病理分級及患者臨床分期相關(guān), MVD隨病理分級的增高而增高;臨床分期Ⅲ~Ⅳ期腫瘤組織的MVD明顯高于Ⅰ~Ⅱ期, MVD或可成為胰腺癌患者預(yù)后指標(biāo)。腫瘤的生長和轉(zhuǎn)移多依賴于腫瘤組織中新生血管的形成 , 而CD105為新生血管所必需的, 有研究顯示, 針對乳腺癌細(xì)胞CD105單克隆抗體已表現(xiàn)出一定的抗腫瘤功效, 且無嚴(yán)重毒副反應(yīng)[13], 提示CD105有可能成為胰腺癌治療的靶點(diǎn), 從而提升胰腺癌的治療效果。

      本研究顯示Annexin A2表達(dá)3+的胰腺導(dǎo)管腺癌組織MVD相對較低, 且Annexin A2表達(dá)2+、+、陰性癌組織之間的MVD差異無統(tǒng)計學(xué)意義, 或許是因?yàn)锳nnexin A2可以促進(jìn)新生血管內(nèi)皮細(xì)胞形成管腔樣結(jié)構(gòu)[14], 使孤立的血管內(nèi)皮細(xì)胞減少, 從而影響MVD計數(shù), 使得Annexin A2表達(dá)3+的腫瘤組織MVD相對較低。當(dāng)然, 其具體的原因及機(jī)制還需進(jìn)一步研究探討。

      參考文獻(xiàn)

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      [2] Vishwanatha J K, Chiang Y, Kumble K D, et al. Enhanced expression of annexin II in human pancreatic carcinoma cells and primary pancreatic cancers.Carcinogenesis,1993,14(12): 2575-2579.

      [3] Domoto T, Miyama Y, Suzuki H, et al. Evaluation of S100A10, annexin II and B-FABP expression as markers for renal cell carcinoma.Cancer science, 2007, 98(1): 77-82.

      [4] Gougos A, Letarte M. Identification of a human endothelial cell antigen with monoclonal antibody 44G4 produced against a pre-B leukemic cell line. J Immunol,1988, 141(6): 1925-1933.

      [5] Tanaka F, Otake Y, Yanagihara K, et al. Correlation between apoptotic index and angiogenesis in non-small cell lung cancer: comparison between CD105 and CD34 as a marker of angiogenesis. Lung Cancer, 2003, 39(3): 289-296.

      [6] Chiang Y,Schneiderman MH,Vishwanatha JK.Annexin II expression is regulated during mammalian cell cycle.Cancer Res, 1993,53(24): 6017-6021.

      [7] Liemann S, Lewit-Bentley A. Annexins:a novel family of calcium-and membrane -bindingproteins in search of a function.Structure, 1995,3(3):233-237.

      [8] Nilius B, Gerke V, Prenen J, et al. Annexin II modulates volume-activated chloride currents in vascular endothelial cells. J Biol Chem, 1996,271(48):30631-30636.

      [9] Ling Q, Jacovina AT, Deora A, et al. Annexin II regulates fibrin homeostasis and neoangiogenesis in vivo.J Clin Invest, 2004, 113(1): 38-48.

      [10] Tanaka F,Otake Y,Yanagihara K,et al. Evaluation of angiogenesis in non - small cell lung cancer : comparison between anti - CD34 antibody and anti - CD105 antibody.Clin Cancer Res,2001(7):3410.

      [11] Fonsatti E,DelVecchio L,AltomonteM,et al. Endoglinn : An accessory component of the TGF-beta 1-binding recep tor-comp lex with diagnostic,prognostic,and bioimmunotherapeutic potential in human malignant .J Cell Physiol,2001,188 (1) :1.

      [12] Kumar S,Ghellal A,Li C,et al. Breast carcinoma: vascular density determined using CD105 antibody correlates with tu-mor prognosis.Cancer Res,1999,59(4):856-561.

      [13] Seon BK,Takahashi N,Haba A,et al. Angiogenesis and metastasis-marker of humantumors.Rinsho Byor, 2001, 49(10):1005-1013.

      [14] Qi Ling,Andrew T,Jacovina,et al. Annexin II regulates fibrin homeostasis and neoangiogenesis in vivo .he Journal of Clinical Investigation,2004, 113(1):38-48.

      本研究顯示Annexin A2表達(dá)3+的胰腺導(dǎo)管腺癌組織MVD相對較低, 且Annexin A2表達(dá)2+、+、陰性癌組織之間的MVD差異無統(tǒng)計學(xué)意義, 或許是因?yàn)锳nnexin A2可以促進(jìn)新生血管內(nèi)皮細(xì)胞形成管腔樣結(jié)構(gòu)[14], 使孤立的血管內(nèi)皮細(xì)胞減少, 從而影響MVD計數(shù), 使得Annexin A2表達(dá)3+的腫瘤組織MVD相對較低。當(dāng)然, 其具體的原因及機(jī)制還需進(jìn)一步研究探討。

      參考文獻(xiàn)

      [1] Gerke V,Moss SE.Annexins: from structure to function. Physiol Rev,2002,82(2): 331-371.

      [2] Vishwanatha J K, Chiang Y, Kumble K D, et al. Enhanced expression of annexin II in human pancreatic carcinoma cells and primary pancreatic cancers.Carcinogenesis,1993,14(12): 2575-2579.

      [3] Domoto T, Miyama Y, Suzuki H, et al. Evaluation of S100A10, annexin II and B-FABP expression as markers for renal cell carcinoma.Cancer science, 2007, 98(1): 77-82.

      [4] Gougos A, Letarte M. Identification of a human endothelial cell antigen with monoclonal antibody 44G4 produced against a pre-B leukemic cell line. J Immunol,1988, 141(6): 1925-1933.

      [5] Tanaka F, Otake Y, Yanagihara K, et al. Correlation between apoptotic index and angiogenesis in non-small cell lung cancer: comparison between CD105 and CD34 as a marker of angiogenesis. Lung Cancer, 2003, 39(3): 289-296.

      [6] Chiang Y,Schneiderman MH,Vishwanatha JK.Annexin II expression is regulated during mammalian cell cycle.Cancer Res, 1993,53(24): 6017-6021.

      [7] Liemann S, Lewit-Bentley A. Annexins:a novel family of calcium-and membrane -bindingproteins in search of a function.Structure, 1995,3(3):233-237.

      [8] Nilius B, Gerke V, Prenen J, et al. Annexin II modulates volume-activated chloride currents in vascular endothelial cells. J Biol Chem, 1996,271(48):30631-30636.

      [9] Ling Q, Jacovina AT, Deora A, et al. Annexin II regulates fibrin homeostasis and neoangiogenesis in vivo.J Clin Invest, 2004, 113(1): 38-48.

      [10] Tanaka F,Otake Y,Yanagihara K,et al. Evaluation of angiogenesis in non - small cell lung cancer : comparison between anti - CD34 antibody and anti - CD105 antibody.Clin Cancer Res,2001(7):3410.

      [11] Fonsatti E,DelVecchio L,AltomonteM,et al. Endoglinn : An accessory component of the TGF-beta 1-binding recep tor-comp lex with diagnostic,prognostic,and bioimmunotherapeutic potential in human malignant .J Cell Physiol,2001,188 (1) :1.

      [12] Kumar S,Ghellal A,Li C,et al. Breast carcinoma: vascular density determined using CD105 antibody correlates with tu-mor prognosis.Cancer Res,1999,59(4):856-561.

      [13] Seon BK,Takahashi N,Haba A,et al. Angiogenesis and metastasis-marker of humantumors.Rinsho Byor, 2001, 49(10):1005-1013.

      [14] Qi Ling,Andrew T,Jacovina,et al. Annexin II regulates fibrin homeostasis and neoangiogenesis in vivo .he Journal of Clinical Investigation,2004, 113(1):38-48.

      本研究顯示Annexin A2表達(dá)3+的胰腺導(dǎo)管腺癌組織MVD相對較低, 且Annexin A2表達(dá)2+、+、陰性癌組織之間的MVD差異無統(tǒng)計學(xué)意義, 或許是因?yàn)锳nnexin A2可以促進(jìn)新生血管內(nèi)皮細(xì)胞形成管腔樣結(jié)構(gòu)[14], 使孤立的血管內(nèi)皮細(xì)胞減少, 從而影響MVD計數(shù), 使得Annexin A2表達(dá)3+的腫瘤組織MVD相對較低。當(dāng)然, 其具體的原因及機(jī)制還需進(jìn)一步研究探討。

      參考文獻(xiàn)

      [1] Gerke V,Moss SE.Annexins: from structure to function. Physiol Rev,2002,82(2): 331-371.

      [2] Vishwanatha J K, Chiang Y, Kumble K D, et al. Enhanced expression of annexin II in human pancreatic carcinoma cells and primary pancreatic cancers.Carcinogenesis,1993,14(12): 2575-2579.

      [3] Domoto T, Miyama Y, Suzuki H, et al. Evaluation of S100A10, annexin II and B-FABP expression as markers for renal cell carcinoma.Cancer science, 2007, 98(1): 77-82.

      [4] Gougos A, Letarte M. Identification of a human endothelial cell antigen with monoclonal antibody 44G4 produced against a pre-B leukemic cell line. J Immunol,1988, 141(6): 1925-1933.

      [5] Tanaka F, Otake Y, Yanagihara K, et al. Correlation between apoptotic index and angiogenesis in non-small cell lung cancer: comparison between CD105 and CD34 as a marker of angiogenesis. Lung Cancer, 2003, 39(3): 289-296.

      [6] Chiang Y,Schneiderman MH,Vishwanatha JK.Annexin II expression is regulated during mammalian cell cycle.Cancer Res, 1993,53(24): 6017-6021.

      [7] Liemann S, Lewit-Bentley A. Annexins:a novel family of calcium-and membrane -bindingproteins in search of a function.Structure, 1995,3(3):233-237.

      [8] Nilius B, Gerke V, Prenen J, et al. Annexin II modulates volume-activated chloride currents in vascular endothelial cells. J Biol Chem, 1996,271(48):30631-30636.

      [9] Ling Q, Jacovina AT, Deora A, et al. Annexin II regulates fibrin homeostasis and neoangiogenesis in vivo.J Clin Invest, 2004, 113(1): 38-48.

      [10] Tanaka F,Otake Y,Yanagihara K,et al. Evaluation of angiogenesis in non - small cell lung cancer : comparison between anti - CD34 antibody and anti - CD105 antibody.Clin Cancer Res,2001(7):3410.

      [11] Fonsatti E,DelVecchio L,AltomonteM,et al. Endoglinn : An accessory component of the TGF-beta 1-binding recep tor-comp lex with diagnostic,prognostic,and bioimmunotherapeutic potential in human malignant .J Cell Physiol,2001,188 (1) :1.

      [12] Kumar S,Ghellal A,Li C,et al. Breast carcinoma: vascular density determined using CD105 antibody correlates with tu-mor prognosis.Cancer Res,1999,59(4):856-561.

      [13] Seon BK,Takahashi N,Haba A,et al. Angiogenesis and metastasis-marker of humantumors.Rinsho Byor, 2001, 49(10):1005-1013.

      [14] Qi Ling,Andrew T,Jacovina,et al. Annexin II regulates fibrin homeostasis and neoangiogenesis in vivo .he Journal of Clinical Investigation,2004, 113(1):38-48.

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