PTEN在肾癌中的表达及与血管生成的关系
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摘要
[背景与目的]
近年来，肾癌的发病率在不断增多，严重危害人类健康。肾癌的发生、发展是一个多阶段、多步骤渐进演化的过程，其发生机制十分复杂，研究表明许多癌基因的激活及抑癌基因的失活参与了这一过程。PTEN为新近发现的一种肿瘤抑制基因，功能十分复杂，具有参与胚胎正常发育，诱导细胞凋亡，引起G_1期细胞阻滞，抑制细胞迁移，铺展和局部粘附功能。新生血管可为肿瘤提供营养物质，也是肿瘤播散转移的前提，同时刺激肿瘤生长，因此新生血管形成是肿瘤发生、发展的关键环节之一。肿瘤新生血管的生成需要PI_3K／AKT信号途径，PTEN可以拮抗PI_3K／AKT途径，从而调节血管内皮生长因子(VEGF)和血小板反应素(TSP-1)的表达而抑制血管形成。有研究表明，脑胶质瘤细胞中PTEN的缺失可以上调VEGF启动子的活性促进肿瘤血管形成。在肾癌中PTEN是否调节VEGF的表达，尚不十分明确。本实验用免疫组化方法，研究正常肾组织和肾癌组织中PTEN、VEGF、微血管密度(MVD)的变化及其相关性，进一步探讨PTEN在肾癌发生发展中的作用，为肾癌的临床诊断和治疗提供理论和依据。
[材料与方法]
实验组为手术切除并经病理确诊的肾癌标本70例，其中男38例，女性32例。病理类型：透明细胞癌40例，颗粒细胞癌30例。有淋巴结转移12例，无淋巴结转移58例。Robson分期：Ⅰ期28例，Ⅱ期30例，Ⅲ期11例，Ⅳ期1例。按Fuhrman标准进行病理分级：透明细胞癌G1、G2、G3、G4级分别为10例、16例、12例、2例；颗粒细胞癌G1、G2、G3、G4级分别为9例、12例、8例、
郑州大学2004年硕士毕业论文
PTEN在肾癌中的表达及与血管生成的关系
1例;同时取14例癌旁非癌肾组织及10例非瘤正常肾组织作对照组。标本用10%
甲醛溶液固定，石蜡包埋，制成厚4um的切片，用SP法对标本进行PTEN、vEGF
及CD3;免疫组化和苏木素一伊红(HE)染色。结果用SPSS10.0软件统计，PTEN，
vEGF蛋白阳性表达率的比较采用x“检验，MvD的分析采用t检验，相关分析
采用spearman相关分析，以P<0.05作为差异有显著性的标准。
【结果]
1.PTEN阳性染色主要表现在细胞核，肾癌组织中阳性表达率为42.9%，显
著低于癌旁非癌肾组织85.7%和正常肾组织100%(P<0.05)。PrEN在透明细胞
癌，颗粒细胞癌阳性表达率分别为45%、40%，二者之间差异无显著性(P>0.os);
PTEN在Gl、GZ、G3一G4级肾透明细胞癌组织中阳性表达率分别为80%、43.8%、
14.3%，在Gl、GZ、G3一G4级肾颗粒细胞癌组织中阳性表达率为77.8%、41.7%、
n.1%，差异均有显著性(P<0.05)。PTEN在I期肾癌组织中阳性表达率为64.3%，H
期肾癌组织中阳性表达率为33.3%，川一Iv期肾癌组织阳性表达率为16.7%，三者
之间差异有显著性(P<0.05)。PTEN在无淋巴结转移的肾癌组织中阳性表达率为
48.3%，显著高于有淋巴结转移肾癌组织(16.7%)(P<0.05)。
2.vEGF阳性染色主要表现在细胞浆，在正常肾组织中阳性表达率为10%，
在肾癌组织中阳性表达率为64.3%，差异有显著性(P<0.05);VEGF在透明细胞
癌中阳性表达率为62.5%，在颗粒细胞癌中阳性表达率为66.7%，差异无显著性
(P>0.05);在Gl、GZ、G3一G4级肾透明细胞癌中阳性表达率分别为40%、56.5%、
92.9%，在Gl、GZ、G3一G4级肾颗粒细胞癌中阳性表达率分别为37.5%、50%、
100%，差异有显著性(P<0.05); vEGF在I期肾癌组织中阳性表达率为39.3%，在H
期肾癌组织中阳性表达率为76.7%，川一Iv期肾癌组织中阳性表达率为91.7%，
差异有显著性(P<0.05);vEGF在有淋巴结转移组肾癌组织中阳性表达率为
91.7%，显著高于无淋巴结转移组(58.6%)(P<0.os)。
3.在正常肾组织中MVD值为20.07士9.32，在肾癌组织中MVD值为53.96
士6.52，差异有显著性(P<0.05);在肾透明细胞癌、颗粒细胞癌中MVD值分
别52.72士5.00，55.62士7.90，差异无显著性(P>0.05);有淋巴结转移组MVD
为59.65士3.85，无淋巴结转移组MVD值为49.69士4.54，差异有显著性(p<0.05).
4.在PTEN表达阳性的30例肾癌标本中，VEGF阳性表达12例，其余18
郑州大学2004年硕士毕业论文
PTEN在肾癌中的表达及与血管生成的关系
例阴性表达;在PTEN表达阴性的40例标本中，VEGF阳性表达33例，其余7
例表达阴性;经Spc盯man相关分析统计学检验，二者呈负相关，差异有显著性
(rs=一0 .896，P<0.05)。
5.vEGF阳性组MvD值为57.31士4.70，显著高于vEGF阴性组的MvD值
47.92士4.74，二者之间差异有显著性(P<0 .05)。经spearman相关分析统计学检
验，vEGF与MVD二者呈正相关，差异有显著性( rs=0.423，P<0.05)。
6.PTEN阴性组MvD值为58.17士4.27，显著高于PTEN阳性组MVD值，
48.34士4.40(P<0 .05)二者之间差异有显著性。经Spcarman相关分析统计学检
验，PTEN与MvD二者呈负相关，差异有显著性(rs=一0.521，P<0 .05)。
[结论」
1.PTEN蛋白在正常肾组织中呈强阳性表达，在肾癌组织中表达减低，提
示它可能在肾癌的发生发展中起重要作用，PTEN的表达与肾癌的组织学类型无
关，与肾癌的病理分级及临床分期和淋巴结转移密切相关。可作为判断肿瘤浸润
进展的指标。
2.vEGF蛋白在正常肾组织中低表?
[Background and objective]
The occurrence of renal carcinoma was increasing notably and it was being severely harmful to humankind health recently. The carcinogenesis of renal tissue is a progression with many steps and phases and its mechanism is rather complex. A lot of reports have showed that activation of oncogenes and inactivation of tumor suppressor genes are involved in renal carcinoma. PTEN is a recently identified tumor suppress gene (TSG), has complexful functions including participating in the embryonic-growth , apoptosis and G1 phase cell block, inhibiting the migration and adherence. The newborn vessels can supply nutrition oxygen for tumors and discharge the metabolic wastes, which provide pathway for tumor metastasis and stimulate the growth of tumor. The angiogenesis is one of crucial steps to the carcinogenesis and tumor progression. Pi3K/AKT signal pathway is essential to vessel formation. PTEN inhibits the PI3K/AKT pathway and it may inhibit the angiogenesis through controlling VEGF and TSP-1. The References of gli
oblastoma tumors reported that loss of PTEN may up-regulate Vascular endothehial growth factor (VEGF) expression, promote angiogene. But it was not clear if PTEN regulate VEGF expression in renal carcinoma. The expressions of PTEN,VEGF and microvessel density (MVD)were analyzed by immunohistochemical method to explore the mechanism of renal carcinogenesis.
[Materials and methods]
70 cases of surgically resected renal tumor tissues were colleced. Among them
male 38 case, female 32 cases .40 cases were renal clear cell carcinoma, and 30 cases were renal granule cell carcinoma, there were lymph nodes metastasis in 12 cases, and no lymph nodes metastasis in 58 cases ; Robson : I 28 cases, II 30 cases, III 11eases, IV 1 cases; Fuhrman grade: renal clear carcinoma Gl 10 cases, G2 16 cases, G3 12 cases , G4 2 cases-, renal granule carcinoma Gl 9 cases, G2 12 cases , G3 8 cases, G4 1 cases; 14 cases adjacent to the carcinoma tissues and 10 cases of normal renal tissues are contrast group. All the tissues were fixed in 10% neutral formalin and embedded in paraffin. Four-um sections were cut from representative blocks of each case. SP immunohistochemical method was performed to detect the expression of PTEN, VEGF and CD34. The data were analyzed by soft Spss 10.0. Chi-square Test and Spearman analyze were used to compare the diffrence of PTEN and VEGF expressions between groups, T tcsl was used lo compove Ihe difference of MVD between groups. P<0.05 was considered as level of teses. [Results]
l.The positive staining of PTEN mainly located in nucleus. The positive rate of PTEN expression was 42.9%in renal carcinoma tissue, the positive expression was 85.7% in renal tissues closely anjacent to carcinoma and the positive expression was 100% in normal renal tissues, a significant difference was observed between the three groups (P<0.05). The positive expression was respectively 45%, 40%in renal clear cell carcinoma, granule cell carcinoma , no significane difference was observed between the two groups(p>0.05).In Gl, G2 > G3-G4 grade renal clear cell carcinoma ,the positive rates were 80%, 43.8%and 14.3% respectively, in Gl , G2, G3-G4 grade renal granule cell carcinoma groups, the positive rates were 77.8% , 41.7%andll.l%respectively,the difference between them was significant(P<0.05);The positive expression was 64.3% in I renal carcinoma tissue,the positive expression was 33.3% in II renal carcinoma tissues and the positive expression was 16.7% in III ~ IV renal carcinoma tissues ,there was signififcant difference between them(p<0.05). In the group with and without lymph nodes metastasis, the positive rate were 16.7% and 48.3% respectively, the difference between them was significane (p<0.05).
2.The positive staining of VEGF mainly located in cytoplasm of renal carcinoma
cells with the positive rate of 64.3%,the positive expression of VEGF in normal tissues was 10% and a significant difference was found beween two groups(p<0.05); The positive expression was respectively 62.
近年来，肾癌的发病率在不断增多，严重危害人类健康。肾癌的发生、发展是一个多阶段、多步骤渐进演化的过程，其发生机制十分复杂，研究表明许多癌基因的激活及抑癌基因的失活参与了这一过程。PTEN为新近发现的一种肿瘤抑制基因，功能十分复杂，具有参与胚胎正常发育，诱导细胞凋亡，引起G_1期细胞阻滞，抑制细胞迁移，铺展和局部粘附功能。新生血管可为肿瘤提供营养物质，也是肿瘤播散转移的前提，同时刺激肿瘤生长，因此新生血管形成是肿瘤发生、发展的关键环节之一。肿瘤新生血管的生成需要PI_3K／AKT信号途径，PTEN可以拮抗PI_3K／AKT途径，从而调节血管内皮生长因子(VEGF)和血小板反应素(TSP-1)的表达而抑制血管形成。有研究表明，脑胶质瘤细胞中PTEN的缺失可以上调VEGF启动子的活性促进肿瘤血管形成。在肾癌中PTEN是否调节VEGF的表达，尚不十分明确。本实验用免疫组化方法，研究正常肾组织和肾癌组织中PTEN、VEGF、微血管密度(MVD)的变化及其相关性，进一步探讨PTEN在肾癌发生发展中的作用，为肾癌的临床诊断和治疗提供理论和依据。
[材料与方法]
实验组为手术切除并经病理确诊的肾癌标本70例，其中男38例，女性32例。病理类型：透明细胞癌40例，颗粒细胞癌30例。有淋巴结转移12例，无淋巴结转移58例。Robson分期：Ⅰ期28例，Ⅱ期30例，Ⅲ期11例，Ⅳ期1例。按Fuhrman标准进行病理分级：透明细胞癌G1、G2、G3、G4级分别为10例、16例、12例、2例；颗粒细胞癌G1、G2、G3、G4级分别为9例、12例、8例、
郑州大学2004年硕士毕业论文
PTEN在肾癌中的表达及与血管生成的关系
1例;同时取14例癌旁非癌肾组织及10例非瘤正常肾组织作对照组。标本用10%
甲醛溶液固定，石蜡包埋，制成厚4um的切片，用SP法对标本进行PTEN、vEGF
及CD3;免疫组化和苏木素一伊红(HE)染色。结果用SPSS10.0软件统计，PTEN，
vEGF蛋白阳性表达率的比较采用x“检验，MvD的分析采用t检验，相关分析
采用spearman相关分析，以P<0.05作为差异有显著性的标准。
【结果]
1.PTEN阳性染色主要表现在细胞核，肾癌组织中阳性表达率为42.9%，显
著低于癌旁非癌肾组织85.7%和正常肾组织100%(P<0.05)。PrEN在透明细胞
癌，颗粒细胞癌阳性表达率分别为45%、40%，二者之间差异无显著性(P>0.os);
PTEN在Gl、GZ、G3一G4级肾透明细胞癌组织中阳性表达率分别为80%、43.8%、
14.3%，在Gl、GZ、G3一G4级肾颗粒细胞癌组织中阳性表达率为77.8%、41.7%、
n.1%，差异均有显著性(P<0.05)。PTEN在I期肾癌组织中阳性表达率为64.3%，H
期肾癌组织中阳性表达率为33.3%，川一Iv期肾癌组织阳性表达率为16.7%，三者
之间差异有显著性(P<0.05)。PTEN在无淋巴结转移的肾癌组织中阳性表达率为
48.3%，显著高于有淋巴结转移肾癌组织(16.7%)(P<0.05)。
2.vEGF阳性染色主要表现在细胞浆，在正常肾组织中阳性表达率为10%，
在肾癌组织中阳性表达率为64.3%，差异有显著性(P<0.05);VEGF在透明细胞
癌中阳性表达率为62.5%，在颗粒细胞癌中阳性表达率为66.7%，差异无显著性
(P>0.05);在Gl、GZ、G3一G4级肾透明细胞癌中阳性表达率分别为40%、56.5%、
92.9%，在Gl、GZ、G3一G4级肾颗粒细胞癌中阳性表达率分别为37.5%、50%、
100%，差异有显著性(P<0.05); vEGF在I期肾癌组织中阳性表达率为39.3%，在H
期肾癌组织中阳性表达率为76.7%，川一Iv期肾癌组织中阳性表达率为91.7%，
差异有显著性(P<0.05);vEGF在有淋巴结转移组肾癌组织中阳性表达率为
91.7%，显著高于无淋巴结转移组(58.6%)(P<0.os)。
3.在正常肾组织中MVD值为20.07士9.32，在肾癌组织中MVD值为53.96
士6.52，差异有显著性(P<0.05);在肾透明细胞癌、颗粒细胞癌中MVD值分
别52.72士5.00，55.62士7.90，差异无显著性(P>0.05);有淋巴结转移组MVD
为59.65士3.85，无淋巴结转移组MVD值为49.69士4.54，差异有显著性(p<0.05).
4.在PTEN表达阳性的30例肾癌标本中，VEGF阳性表达12例，其余18
郑州大学2004年硕士毕业论文
PTEN在肾癌中的表达及与血管生成的关系
例阴性表达;在PTEN表达阴性的40例标本中，VEGF阳性表达33例，其余7
例表达阴性;经Spc盯man相关分析统计学检验，二者呈负相关，差异有显著性
(rs=一0 .896，P<0.05)。
5.vEGF阳性组MvD值为57.31士4.70，显著高于vEGF阴性组的MvD值
47.92士4.74，二者之间差异有显著性(P<0 .05)。经spearman相关分析统计学检
验，vEGF与MVD二者呈正相关，差异有显著性( rs=0.423，P<0.05)。
6.PTEN阴性组MvD值为58.17士4.27，显著高于PTEN阳性组MVD值，
48.34士4.40(P<0 .05)二者之间差异有显著性。经Spcarman相关分析统计学检
验，PTEN与MvD二者呈负相关，差异有显著性(rs=一0.521，P<0 .05)。
[结论」
1.PTEN蛋白在正常肾组织中呈强阳性表达，在肾癌组织中表达减低，提
示它可能在肾癌的发生发展中起重要作用，PTEN的表达与肾癌的组织学类型无
关，与肾癌的病理分级及临床分期和淋巴结转移密切相关。可作为判断肿瘤浸润
进展的指标。
2.vEGF蛋白在正常肾组织中低表?
[Background and objective]
The occurrence of renal carcinoma was increasing notably and it was being severely harmful to humankind health recently. The carcinogenesis of renal tissue is a progression with many steps and phases and its mechanism is rather complex. A lot of reports have showed that activation of oncogenes and inactivation of tumor suppressor genes are involved in renal carcinoma. PTEN is a recently identified tumor suppress gene (TSG), has complexful functions including participating in the embryonic-growth , apoptosis and G1 phase cell block, inhibiting the migration and adherence. The newborn vessels can supply nutrition oxygen for tumors and discharge the metabolic wastes, which provide pathway for tumor metastasis and stimulate the growth of tumor. The angiogenesis is one of crucial steps to the carcinogenesis and tumor progression. Pi3K/AKT signal pathway is essential to vessel formation. PTEN inhibits the PI3K/AKT pathway and it may inhibit the angiogenesis through controlling VEGF and TSP-1. The References of gli
oblastoma tumors reported that loss of PTEN may up-regulate Vascular endothehial growth factor (VEGF) expression, promote angiogene. But it was not clear if PTEN regulate VEGF expression in renal carcinoma. The expressions of PTEN,VEGF and microvessel density (MVD)were analyzed by immunohistochemical method to explore the mechanism of renal carcinogenesis.
[Materials and methods]
70 cases of surgically resected renal tumor tissues were colleced. Among them
male 38 case, female 32 cases .40 cases were renal clear cell carcinoma, and 30 cases were renal granule cell carcinoma, there were lymph nodes metastasis in 12 cases, and no lymph nodes metastasis in 58 cases ; Robson : I 28 cases, II 30 cases, III 11eases, IV 1 cases; Fuhrman grade: renal clear carcinoma Gl 10 cases, G2 16 cases, G3 12 cases , G4 2 cases-, renal granule carcinoma Gl 9 cases, G2 12 cases , G3 8 cases, G4 1 cases; 14 cases adjacent to the carcinoma tissues and 10 cases of normal renal tissues are contrast group. All the tissues were fixed in 10% neutral formalin and embedded in paraffin. Four-um sections were cut from representative blocks of each case. SP immunohistochemical method was performed to detect the expression of PTEN, VEGF and CD34. The data were analyzed by soft Spss 10.0. Chi-square Test and Spearman analyze were used to compare the diffrence of PTEN and VEGF expressions between groups, T tcsl was used lo compove Ihe difference of MVD between groups. P<0.05 was considered as level of teses. [Results]
l.The positive staining of PTEN mainly located in nucleus. The positive rate of PTEN expression was 42.9%in renal carcinoma tissue, the positive expression was 85.7% in renal tissues closely anjacent to carcinoma and the positive expression was 100% in normal renal tissues, a significant difference was observed between the three groups (P<0.05). The positive expression was respectively 45%, 40%in renal clear cell carcinoma, granule cell carcinoma , no significane difference was observed between the two groups(p>0.05).In Gl, G2 > G3-G4 grade renal clear cell carcinoma ,the positive rates were 80%, 43.8%and 14.3% respectively, in Gl , G2, G3-G4 grade renal granule cell carcinoma groups, the positive rates were 77.8% , 41.7%andll.l%respectively,the difference between them was significant(P<0.05);The positive expression was 64.3% in I renal carcinoma tissue,the positive expression was 33.3% in II renal carcinoma tissues and the positive expression was 16.7% in III ~ IV renal carcinoma tissues ,there was signififcant difference between them(p<0.05). In the group with and without lymph nodes metastasis, the positive rate were 16.7% and 48.3% respectively, the difference between them was significane (p<0.05).
2.The positive staining of VEGF mainly located in cytoplasm of renal carcinoma
cells with the positive rate of 64.3%,the positive expression of VEGF in normal tissues was 10% and a significant difference was found beween two groups(p<0.05); The positive expression was respectively 62.
引文
1. Steck PA, Pershouse MA, Jasser SA, et al. Indentification of a candidate tumor suppressor gene, MMACI,al chromosome 10q23.3,which is mutated in multiple advanced Cancers.Nat Genet, 1997;15:357-362.
2. Li J, Yen C, Liaw D, et al. PTEN, a putative protein tyrosine phosphatases gene mutated in humam brain, breast, and prostate cancer. Science, 1997,275:1943-1947.
3. Depowski PL, Rosenthal SI, Ross JS. Loss of expression of the PTEN gene protein product is associated with poor outcome in breast canccr. Mod Pathol,2001; 14(7): 672-676.
4. Hopin K.A surprising function for the PTEN tumor suppressor. Sciene, 1998;282:1027-1030.
5. Tamura M, Gu J, Matasumoto K, et al. Inhibition of cell migration,spreading, and focal adhesions by tumor suppressor PTEN. Sciene, 1998;280:1614-1617.
6. Hanahan D, Folkman J. Patterns and emerging mechanisms of the angiogcnesis switch during tumorigenesis. Cell, 1996;86:353-364.
7. Bates DO, Heald RI, Caeey FE,et al.VEGF increases Rana vascular permeability and compliance by different signaling pathways. J Physiol,2001;53:263.
8. Jiang BH, Zheng JZ, Aoki M, et al. Phosphatidylinositol 3-kinase signaling mediates angiogenesis and expression of vascular endothelial growth factor in endothelial cells. Proc NatI Acad Sci USA, 2000; 97(4): 1749-1753.
9. Gomez-Manzano C, Fueyo J, Jiang H, et al. Mechanisms underlying PTEN regulation of vascular endothelial growth factor and angiogenesis. Ann Neurol, 2003 ;53(1):109-117.
10. Gulderg P, Straten PT, Birck A, et al. Disruption to the PTEN/MMAC1 gene by deletion or mutation is a frequent event in malignant melanoma. Cancer Res, 1999;57:3660-3663.
11. Hwang PH, Yi HF, Kim DS, et al. Suppression of tumorgenesis and metastasis in B16F10 cells by PTEN/MMAC1/TEP1 gene. Cancer Lett, 2001;172(1): 83-91.
12. Pore N, Liu S, Haas-Kogan DA, et al. PTEN mutation and epidermal growth factor receptor activation regulate vascular endothelial growth factor (VEGF) Mrna expression in human glioblastoma cells by transactivating the proximal VEGF promoter.Cancer Res,2003 ;63(1):236-241.
13. Weinder N, Semple J, Welch WR, et al. Tumor angiogenesis and metastasis relation in invasive breast carcinoma. N Eng J Med, 1991;324:1-8.
14. Wang SI, Parsons R, Ittmann M, et al. Homozygous deletion of the PTEN tumor suppressor gene in a subset of prostate adcnocarcinomas. Clin Cancer Res, 1998;4:811-815.
15. Cairs P, Evren E, Okami K, et al. Point mutation and homozygous deletion of PTEN/MMAC1 in primary bladder cancers. Oncogcne, 1998;16(24):3215-3218.
16. Mutter GL, Lin MC, Fitzgerald JT, et al. Altered PTEN expression of a diagnostic Marker for the earliest endometrial precancers. Nat Cancer Inst, 2000;7(92):924-930.
17. MCMenamin ME, Soung P, Perera S, et al. Loss of PTEN Expression in paraffin-embedded Primary Prostate cancer correlates with High Gleason Score and Advanced stage. Cancer Res,1999;59:4291-4296.
18. Whang YE, Wu X, Suzuki H, et al. Inactivation of the tumor suppressor PTEN/MMAC1 in advanced human prostate cancer through loss of expression. Proc Nati Acad Sci USA, 1998;95: 5246-5250.
19. Sano T, Lin H, Chen X, et al. Differential Expression of MMAC1/PTEN in Glioblastoma Multiforme: Relationship to localization and Prognosis. Cancer Res, 1999; 59(8): 1820-1824.
20. 李金雨，曹润福，汪洪等，抑癌基因PTEN／MMAC1／TEP1在肾癌组织中的表达及其与细胞增殖和凋亡的关系．癌症杂志，2003；22(6)：607-611．
21. Holash J, Maisonpierre PC, Compton D, et al. vessel cooption, regression, and growth in tumor mediated by angiopoietins and VEGE Scicnce,1999;284:1994-1998.
22. Neufled G, Cohen PC, Gengrinovitch S,et al. Vascular endothelial growth factor(VEGF)and its receptors.FASEB J,1999;13(1):9-22.
23. 石歧兴 白晓军 关宏华等．肾癌组织中血管内皮生长因子表达的意义．实用癌症杂志，2001；16(3)：239-241．
24. Zhang X, Yamashita M,Uctsuki H et al.Angiogenesis in renal cell carcinoma:Evaluation of microvessel density, vascular endothelial growth factor and matrix metalloproteinases. Int J Urology, 2002;9(9):509-514.
25. Dekel Y, Koren R, Kugel V, et al.Significance of angiogenesis and microvascular invasion in renal cell carcinoma. Pathol Oncol Res,2002;8(2):129-132.
26. Kirkali Z, Yorukoglu K, Ozkara E.Proliferative activity, angiogenesis and nuclear morphometry in renal cell carcinoma.Int J Urology,2001;8(12):697-703.
27. SuzukiK, Morita T, Hashimoto S,et al.Thymidine hosphorylase/platelet-derivcd endothelial cell growth factor(PD-ECGF) associated with prognosis in renal cell carcinoma.Urology Res ,2001;29(1):7-12.
28. Wen SH, Stolarov J, Myers MP, et al. PTEN controls tumor-induced angiogcnesis. Proc Nati Acad Sci USA, 2001;98(8): 4622-4627.
29. Plouet J, Moro F, Bertagnolli S,et al. Extracellular cleavage of the vascular endothelial growth factor 189-amino acid form by urokinase is required for its mitogenic effect.J Boil Chem,1997;272:13390-13396.
30. Giri D, Ittmann M. Inactivation of the PTEN tumor suppressor gene is associated with increased angiogenesis in clinically localized prostate carcinoma. Hum Pathol,1999;30(4): 419-424.
31. Risan W.What,if anything, is an angiogerenic factor?Cancer Metastasis Rev, 1996;15(1): 149-151.
32. Kroll J,Waltenberger J.Regulation of the endothelid function and angiogenesis by vascular endothetial growth-A(vegf-A).Zkardiol,2000;89(3):206-218.
33. Folkman J.What is the evidence that tumors are angiogenesis dependent. J Natl Cancer Inst, 1990;82(1):4-6.
34. Huang J, Kontos CD. PTEN modulates vascular endothelial growth factor-mediated signaling and angiogenic effects. J Biol Chem,2002;277(13): 10760-10766.
35. Koul D, Shen R, Garyali A, et al. MMAC1/PTEN tumor suppressor gene regulates
VEGF-mediated angiogenesis in prostate cancer.Int J Oncol,2002;21(3): 469-475.
2. Li J, Yen C, Liaw D, et al. PTEN, a putative protein tyrosine phosphatases gene mutated in humam brain, breast, and prostate cancer. Science, 1997,275:1943-1947.
3. Depowski PL, Rosenthal SI, Ross JS. Loss of expression of the PTEN gene protein product is associated with poor outcome in breast canccr. Mod Pathol,2001; 14(7): 672-676.
4. Hopin K.A surprising function for the PTEN tumor suppressor. Sciene, 1998;282:1027-1030.
5. Tamura M, Gu J, Matasumoto K, et al. Inhibition of cell migration,spreading, and focal adhesions by tumor suppressor PTEN. Sciene, 1998;280:1614-1617.
6. Hanahan D, Folkman J. Patterns and emerging mechanisms of the angiogcnesis switch during tumorigenesis. Cell, 1996;86:353-364.
7. Bates DO, Heald RI, Caeey FE,et al.VEGF increases Rana vascular permeability and compliance by different signaling pathways. J Physiol,2001;53:263.
8. Jiang BH, Zheng JZ, Aoki M, et al. Phosphatidylinositol 3-kinase signaling mediates angiogenesis and expression of vascular endothelial growth factor in endothelial cells. Proc NatI Acad Sci USA, 2000; 97(4): 1749-1753.
9. Gomez-Manzano C, Fueyo J, Jiang H, et al. Mechanisms underlying PTEN regulation of vascular endothelial growth factor and angiogenesis. Ann Neurol, 2003 ;53(1):109-117.
10. Gulderg P, Straten PT, Birck A, et al. Disruption to the PTEN/MMAC1 gene by deletion or mutation is a frequent event in malignant melanoma. Cancer Res, 1999;57:3660-3663.
11. Hwang PH, Yi HF, Kim DS, et al. Suppression of tumorgenesis and metastasis in B16F10 cells by PTEN/MMAC1/TEP1 gene. Cancer Lett, 2001;172(1): 83-91.
12. Pore N, Liu S, Haas-Kogan DA, et al. PTEN mutation and epidermal growth factor receptor activation regulate vascular endothelial growth factor (VEGF) Mrna expression in human glioblastoma cells by transactivating the proximal VEGF promoter.Cancer Res,2003 ;63(1):236-241.
13. Weinder N, Semple J, Welch WR, et al. Tumor angiogenesis and metastasis relation in invasive breast carcinoma. N Eng J Med, 1991;324:1-8.
14. Wang SI, Parsons R, Ittmann M, et al. Homozygous deletion of the PTEN tumor suppressor gene in a subset of prostate adcnocarcinomas. Clin Cancer Res, 1998;4:811-815.
15. Cairs P, Evren E, Okami K, et al. Point mutation and homozygous deletion of PTEN/MMAC1 in primary bladder cancers. Oncogcne, 1998;16(24):3215-3218.
16. Mutter GL, Lin MC, Fitzgerald JT, et al. Altered PTEN expression of a diagnostic Marker for the earliest endometrial precancers. Nat Cancer Inst, 2000;7(92):924-930.
17. MCMenamin ME, Soung P, Perera S, et al. Loss of PTEN Expression in paraffin-embedded Primary Prostate cancer correlates with High Gleason Score and Advanced stage. Cancer Res,1999;59:4291-4296.
18. Whang YE, Wu X, Suzuki H, et al. Inactivation of the tumor suppressor PTEN/MMAC1 in advanced human prostate cancer through loss of expression. Proc Nati Acad Sci USA, 1998;95: 5246-5250.
19. Sano T, Lin H, Chen X, et al. Differential Expression of MMAC1/PTEN in Glioblastoma Multiforme: Relationship to localization and Prognosis. Cancer Res, 1999; 59(8): 1820-1824.
20. 李金雨，曹润福，汪洪等，抑癌基因PTEN／MMAC1／TEP1在肾癌组织中的表达及其与细胞增殖和凋亡的关系．癌症杂志，2003；22(6)：607-611．
21. Holash J, Maisonpierre PC, Compton D, et al. vessel cooption, regression, and growth in tumor mediated by angiopoietins and VEGE Scicnce,1999;284:1994-1998.
22. Neufled G, Cohen PC, Gengrinovitch S,et al. Vascular endothelial growth factor(VEGF)and its receptors.FASEB J,1999;13(1):9-22.
23. 石歧兴 白晓军 关宏华等．肾癌组织中血管内皮生长因子表达的意义．实用癌症杂志，2001；16(3)：239-241．
24. Zhang X, Yamashita M,Uctsuki H et al.Angiogenesis in renal cell carcinoma:Evaluation of microvessel density, vascular endothelial growth factor and matrix metalloproteinases. Int J Urology, 2002;9(9):509-514.
25. Dekel Y, Koren R, Kugel V, et al.Significance of angiogenesis and microvascular invasion in renal cell carcinoma. Pathol Oncol Res,2002;8(2):129-132.
26. Kirkali Z, Yorukoglu K, Ozkara E.Proliferative activity, angiogenesis and nuclear morphometry in renal cell carcinoma.Int J Urology,2001;8(12):697-703.
27. SuzukiK, Morita T, Hashimoto S,et al.Thymidine hosphorylase/platelet-derivcd endothelial cell growth factor(PD-ECGF) associated with prognosis in renal cell carcinoma.Urology Res ,2001;29(1):7-12.
28. Wen SH, Stolarov J, Myers MP, et al. PTEN controls tumor-induced angiogcnesis. Proc Nati Acad Sci USA, 2001;98(8): 4622-4627.
29. Plouet J, Moro F, Bertagnolli S,et al. Extracellular cleavage of the vascular endothelial growth factor 189-amino acid form by urokinase is required for its mitogenic effect.J Boil Chem,1997;272:13390-13396.
30. Giri D, Ittmann M. Inactivation of the PTEN tumor suppressor gene is associated with increased angiogenesis in clinically localized prostate carcinoma. Hum Pathol,1999;30(4): 419-424.
31. Risan W.What,if anything, is an angiogerenic factor?Cancer Metastasis Rev, 1996;15(1): 149-151.
32. Kroll J,Waltenberger J.Regulation of the endothelid function and angiogenesis by vascular endothetial growth-A(vegf-A).Zkardiol,2000;89(3):206-218.
33. Folkman J.What is the evidence that tumors are angiogenesis dependent. J Natl Cancer Inst, 1990;82(1):4-6.
34. Huang J, Kontos CD. PTEN modulates vascular endothelial growth factor-mediated signaling and angiogenic effects. J Biol Chem,2002;277(13): 10760-10766.
35. Koul D, Shen R, Garyali A, et al. MMAC1/PTEN tumor suppressor gene regulates
VEGF-mediated angiogenesis in prostate cancer.Int J Oncol,2002;21(3): 469-475.