Relationship between the expression of iNOS,VEGF,tumor angiogenesis and gastric cancer

AIM：To in vestigate the relationship between the expression of inducible nitric oxide synthase(iNOS),vascular endothelial growth factor(VEGF),the microvascular density(MVD)and the pathological features and clinical staging of gastric cancer.METHODS:Immunohistochemical staining was used for detecting the expression of iNOS and VEGFin46resected specimens of gastric carcinoma;the monoclonal antibody against CD34 was used for displaying vascular endothelial cells,and MVD was detected by counting of CD34-positive vascular endothelial cells.RESULTS:Of 46resected specimens of gastric carcinoma,the rates of expressions of iNOS and VEGF were 58.70%and76.09%,respectively,and MVDaveraged55.59±19.39,Judged by the standard TNM criteria,the rate of expression of iNOS in stageⅣ（84.46%)was higher than those in stageⅠ,Ⅱ,Ⅲ（Fish exact probabilities test,P=0.019,0.023and 0.033,respectively);the rates of expression of VEGFin stage Ⅲ,Ⅳ（76.0%,92.31%,respectively)were higher than those in stageⅠ,Ⅱ（Fis exact probabilities test,P=0.031,0.017,0.022and0.019).MVDs in stageⅢ,Ⅳ（64.72±14.96,67.09±18.29,respectively)were higher than those in stageⅠ,Ⅱ(t\2.378,4.015,2.503and2.450,P<0.05,P<0.001,P<0.001,P<0.05,respectively),In37gastric carcinoma specimens with lymph node metastasis,MVD(68.69±18.07)and the rates of expression of iNOS and VEGF(70.27%,83.78%,respectively)were higher than those in the specimens with absence of metastasis(t=2.205,X^2=6.3587,X^2=6.2584,P<0.01,P<0.05,P<0.05,respectively),MVD and the expressions of iNOS and EGF were not correlated to the location,size or grade of tumor,nor with the depth of invasion of tumor;MVDs in the positive iNOS and VEGF specimens(59.88±18.02,58.39±17.73,repectively)were higher than those in the negative iNOS and VEGF specimens(X^2=6.3587and 6.1574,P<0.05,P<0.05,respectively);thus the expressions of iNOS and VEGF was correlated to MVD,but the expression of iNOS was not correlated to that of VEGF,In addition.of the 46 surviving patients,the 5-year survival rate of patients with positive iNOS or VEGF tumors was significantly less than that of patients with negative iNOS-or VEGF tumors(X^2=4.3842and 5.4073,P<0.05,P<0.05.respectively).CONCLUSION:The expressions of iNOS and VEGF are colosely related to tumor angiogenesis,and are involved in the advancement and the lymph node metastasis;thusMVD and the expressions of iNOS and EGF may serve indexes for evaluating staging of gastric carcinoma and forecasting its risk of metastasis,which will help establish a comprehensive therapeutical measure of post-operative patients and provide a new approach to tumor therapy.     

肾上腺皮质肿瘤VEGF、TSP-1的表达及其与微血管密度的关系

目的 研究血管内皮生长因子(VEGF)、血小板反应素1(TSP-1)在肾上腺皮质肿瘤中的表达及其与微血管密度(MVD)的关系.方法 采用免疫组化方法检测13例肾上腺皮质癌、30例肾上腺皮质腺瘤和7例正常肾上腺组织中VEGF、TSP-1及MVD(以CD34为标记)的表达.结果 肾上腺皮质癌VEGF表达高于肾上腺皮质腺瘤,差异有统计学意义(P＜0.01);TSP-1在正常肾上腺皮质表达明显高于肾上腺皮质癌(P＜0.01);MVD在肾上腺皮质癌为(107.67±11.91)/视野,显著高于肾上腺皮质腺瘤(59.46±21.31)/视野和正常肾上腺皮质组织(25.10±3.94)/视野(均P＜0.01);VEGF的表达与MVD呈正相关(P＜0.01),TSP-1的表达与MVD呈负相关(P＜0.01),VEGF与TSP-1的表达呈负相关(P＜0.01).结论 VEGF和TSP-1在肾上腺皮质肿瘤的表达失衡是其肿瘤血管异常生成的重要原因.     

Relationship between the expression of iNOS, VEGF, tumor angiogenesis and gastric cancer

AIM: To investigate the relationship between theexpression of inducible nitric oxide synthase(iNOS),vascular endothelial growth factor(VEGF),themicrovascular density(MVD) and the pathologicalfeatures and clinical staging of gastric cancerMETHODS: Immunohistochemical staining was used fordetecting the expression of iNOS and VEGF in 46resected specimens of gastric carcinoma; themonoclonal antibody against CD34 was used fordisplaying vascular endothelial cells, and MVD wasdetected by counting of CD34-positive vascularendothelial cells.RESULTS: Of 46 resected specimens of gastriccarcinoma,the rates of expressions of iNOS and VEGFwere 58.70% and 76.09%, respectively,and MVDaveraged 55.59± 19.39.Judged by the standard TNMcriteria, the rate of expression of iNOS in stage ⅣV(84.46%) was higher than those in stage Ⅰ, Ⅱ, Ⅲ(Fishexact probabilities test, P=0.019,0.023 and 0.033,respectively);the rates of expression of VEGF in stageⅢ, Ⅳ (76.0%,92.31%, respectively) were higher thanthose in stage Ⅰ, Ⅱ (Fish exact probabilities test,P=0.031,0.017,0.022 and 0.019). MVDs in stage Ⅲ, Ⅳ(64.72 ± 14.96,67.09± 18.29,respectively) werehigher than those in stage Ⅰ ,Ⅱ(t=2.378,4.015,2.503and 2.450,P＜0.05,P＜0.001,P＜0.001,P＜0.05,respectively). In 37 gastric carcinoma specimens withlymph node metastasis, MVD(68.69±18.07) and therates of expression of iNOS and VEGF(70.27%,83.78%,respectively) were higher than those in the specimenswith absence of metastasis (t=2.205, x2=6.3587,x2=6.2584,P＜0.01, P＜0.05,P＜0.05, respectively). MVDand the expressions of iNOS and VEGF were notcorrelated to the location ,size or grade of tumor, nor withthe depth of invasion of tumor; MVDs in the positive iNOSand VEGF specimens(59.88±18.02,58.39±17.73,respectively) were higher than those in the negative iNOSand VEGF specimens (x2=6.3587 and 6.1574, P＜0.05,P＜0.05,respectively) ;thus the expressions of iNOS andVEGF was correlated to MVD, but the expression of iNOSwas not correlated to that of VEGF. In addition,of the46 surviving patients, the 5-year survival rate ofpatients with positive iNOS or VEGF tumors wassignificantly less than that of patients with negativeiNOS-or VEGF tumors(x2=4.3842 and 5.4073,P＜0.05,P＜ 0.05,respectively).CONCLUSION: The expressions of iNOS and VEGF areclosely related to tumor angiogenesis, and are involvedin the advancement and the lymph node metastasis;thus MVD and the expressions of iNOS and VEGF mayserve indexes for evaluating staging of gastriccarcinoma and forecasting its risk of metastasis, whichwill help establish a comprehensive therapeuticalmeasure of post-operative patients and provide a newapproach to tumor therapy.     

Inactivation of PTEN is associated with increased angiogenesis and VEGF overexpression in gastric cancer

AIM: To investigate the expression of PTEN/MMAC1/TEP1 and vascular endothelial growth factor (VEGF), their roles in biologic behavior and angiogenesis and their association in gastric cancer.METHODS: Immunohistochemical staining was used to evaluate the expression of PTEN, VEGF and microvascular density (MVD) on paraffin-embedded sections in 70 patients with primary gastric cancer and 24 patients with chronic superficial gastritis (CSG). Expression of PTEN, VEGF and MVD were compared with clinicopathological features of gastric cancer. The relationship between expression of PTEN, VEGF and MVD as well as the relationship between PTEN and VEGF expression in caner cells were investigated. RESULTS: PTEN expression significantly decreased (t= 3.98, P<0.01) whereas both VEGF expression and MVD significant increased (t = 4.29 and 4.41, respectively, both P<0.01) in gastric cancer group compared with CSG group. PTEN expression was significantly down-regulated (t=1.95, P<0.05) whereas VEGF expression (t = 2.37, P<0.05) and MVD (t= 3.28, P<0.01) was significantly up-regulated in advanced gastric cancer compared with early-stage gastric cancer. PTEN expression in gastric cancer showed a negative association with lymph node metastasis (t= 3.91, P<0.01), invasion depth (t= 1.95, P<0.05) and age (t= 4.69, P<0.01). MVD in PTEN-negative gastric cancer was significantly higher than that in PTEN-positive gastric cancer (t=3.69, P<0.01), and there was a negative correlation betweenPTEN expression and MVD (γ=-0.363, P<0.05). VEGF expression was positively associated with invasion depth (especially with serosa invasion, t = 4.69, P<0.01), lymph node metastasis (t= 2.31, P<0.05) and TNM stage (t= 3.04, P<0.01). MVD in VEGF-positive gaslyic cancer was significantly higher than that in VEGF-negative gastric cancer (t=4.62, P<0.01), and there was a positive correlation between VEGF expression of and MVD (y = 0.512, P<0.05). VEGF expression in PTEN-negative gaslyic cancer was significantly stronger than that in PTEN-positive gastric cancer (t=2.61, P<0.05), and there was a significantly negative correlation between the expression of VEGF and PTEN (γ=-0.403, P<0.05).CONCLUSION: Our results imply that inactivation of PTEN gene and over-expression of VEGF contribute to the neovascularization and progression of gastric cancer. PTEN-related angiogenesis might be attributed to its up-regulation of VEGF expression. PTEN and VEGF could be used as the markers reflecting the biologic behaviors of tumor and viable targets in therapeutic approaches to inhibit angiogenesis of gastric cancers.     

Role of transforming growth factor-beta1-smad signal transduction pathway in patients with hepatocellular carcinoma

AIM:To explore the role of transforming growth factor-beta1(TGF-β1)-smad signal transduction pathway inpatients with hepatocellular carcinoma.METHODS:Thirty-six hepatocellular carcinoma speci-mens were obtained from Qidong Liver Cancer Instituteand Department of Pathology of the Second AffiliatedHospital of Nanjing Medical University.All primary anti-bodies(polyclonal antibodies)to TGF-β1,type Ⅱ Trans-forming growth factor-beta receptor(TβR-Ⅱ),nuclear fac-tor-kappaB(NF-KB),CD34,smad4 and smad7,secondaryantibodies and immunohistochemical kit were purchasedfrom Zhongshan Biotechnology Limited Company(Bei-jing,China).The expressions of TGF-β1,TβR-Ⅱ,NF-kB,smad4 and smad7 proteins in 36 specimens ofhepatocellular carcinoma(HCC)and its adjacent tissuewere separately detected by immunohistochemistry toobserve the relationship between TGF-β1 and TβR-Ⅱ,between NF-kB and TGF-β1,between smad4 and smad7and between TGF-β1 or TβR-Ⅱ and microvessel density(MVD).MVD was determined by labelling the vesselendothelial cells with CD34.RESULTS:The expression of TGF-β1,smad7 and MVDwas higher in HCC tissue than in adjacent HCC tissue(P<0.01,P<0.05,P<0.01 respectively).The expressionof TβR-Ⅱ and smad4 was lower in HCC tissue than in its adjacent tissue(P<0.01,P<0.05 respectively).Theexpression of TGF-β1 protein and NF-kB protein wasconsistent in HCC tissue.The expression of TGF-β1 andMVD was also consistent in HCC tissue.The expressionof TβR-Ⅱ was negatively correlated with that of MVD inHCC tissue.CONCLUSION:The expressions of TGF-β1,TβR-Ⅱ,NF-kB,smad4 and smad7 in HCC tissue,which are ma-jor up and down stream factors of TGF-β1-smad signaltransduction pathway,are abnormal.These factors areclosely related with MVD and may play an important rolein HCC angiogenesis.The inhibitory action of TGF-β1 isweakened in hepatic carcinoma cells because of abnor-mality of TGF-β1 receptors(such as TβR-Ⅱ)and postre-ceptors(such as smad4 and smad7).NF-kB may causeactivation and production of TGF-β1.     

Correlation of integrin β3 mRNA and vascular endothelial growth factor protein expression profiles with the clinicopathological features and prognosis of gastric carcinoma

AIM： To investigate integrin β3 mRNA and vascular endothelial growth factor （VEGF） protein expression in gastric carcinoma, and its correlation with microvascular density, growth-pattern, invasion, metastasis and prognosis. METHODS： In situ hybridization（ISH） of integrin β3 mRNA and immunohistochemistry of VEGF and CD34 protein were performed on samples from 118 patients with gastric cancer. RESULTS： The positive rate of integrin β3 mRNA in non-tumor gastric mucosa （20%） was significantly lower than that of the gastric cancer tissue （52.5%, X^2 = 10.20, P 〈 0.01）. In patients of infiltrating type, stage T3-T4, vessel invasion, lymphatic metastasis, hepatic or peritoneal metastasis, the positive expression rates of integrin β3 mRNA were significantly higher than those in patients of expanding type （P 〈 0.01）, stage T1-T2 （P 〈 0.01）, non-vessel invasion （P 〈 0.01）, without lymphatic metastasis （P 〈 0.01）, without hepatic and peritoneal metastasis （P 〈 0.01）, respectively. In patients of infiltrating type, stage T3-T4, vessel invasion, lymphatic metastasis, hepatic or peritoneal metastasis, the positive expression rates of VEGF protein were significantly higher than those in patients of expanding type （P 〈 0.01）, stage T1-T2 （P 〈 0.01）, non-vessel invasion （P 〈 0.01）, without lymphatic metastasis （P 〈 0.01）, without hepatic and peritoneal metastasis （P 〈 0.01）, respectively. In patients of infiltrating type, stage T3-T4, vessel invasion, lymphatic metastasis, hepatic or peritoneal metastasis, the mean MVD were significantly higher than those in patients of expanding type （P 〈 0.01）, stage T1-T2 （P 〈 0.01）, non-vessel invasion （P 〈 0.01）, without lymphatic metastasis （P 〈 0.01）, without hepatic and peritoneal metastasis （P 〈 0.01）, respectively. It was found that the positive expression rate of integrin β3 mRNA was positively related to that of VEGF protein （P 〈 0.01） and MVD （P 〈 0.05）,     

胃癌组织MMP-10和VEGF表达与血管生成的关系

目的:研究胃癌组织中基质金属蛋白酶- 10(MMP-10)、血管内皮生长因子(VEGF)和微血管密度(MVD)表达变化及其与肿瘤临床病理特征之间的关系.方法:以CD31作为MVD指标,应用免疫组化法检测60例胃癌组织和60例距病灶5 cm以上的正常组织中的MMP-10、VEGF和CD31的表达并对结果进行分析.结果:60例胃癌组织中的MMP-10、VEGF的表达阳性率分别为81.7%、76.7%,明显高于正常组织的表达阳性率11.7%、8.3%,两者差异有统计学意义(P<0.05).MMP-10、VEGF的表达与MVD、与肿瘤的分化程度、TNM分型、淋巴结的转移、浸润程度有关.结论:MMP-10、VEGF胃癌组织中高表达与胃癌侵袭转移、血管生成密切相关,可作为判断胃癌侵袭转移及预后的重要指标.     

Correlative studies on uPA mRNA and uPAR mRNA expression with vascular endothelial growth factor, microvessel density, progression and survival time of patients with gastric cancer

AIM: TO investigate the correlations between the expression of urokinase-type plasminogen activator (uPA) mRNA, uPA receptor (uPAR) mRNA and vascular endothelial growth factor (VEGF) protein and clinicopathologic features, microvessel density (MVD) and survival time. METHODS: In situ hybridization and immuno-histochemistry techniques were used to study the expressions of uPA mRNA, uPAR mRNA, VEGF and CD34 protein in 105 gastric carcinoma specimens. RESULTS: Expressions of uPA mRNA, uPAR mRNA and VEGF protein were observed in 61 (58.1%) cases, 70 (66.7%) cases and 67 (63.8%) cases, respectively. The uPA mRNA and uPAR mRNA positive expression rates in infiltrating-type cases (73.7%, 75.4%), stageⅢ-Ⅳ(72.1%, 75.4%), vessel invasion (63.2%, 69.9%), lymphatic metastasis (67.1%, 74.4%) and distant metastasis (88.1%, 85.7%) were significantly higher than those of the expanding-type (X2= 15.57, P= 0.001; X2=6.91, P=0.046), stageⅠ-Ⅱ(X2 = 19.22, P = 0.001; X2= 16.75, P= 0.001), non-vessel invasion (X2 = 11.92, P = 0.006; X2 = 14.15, P = 0.002), non-lymphatic metastasis (X2 = 28.41, P = 0.001; X2= 22.5, P=0.005) and non-distant metastasis (X2 = 12.32, P= 0.004; X2= 17.42, P = 0.002; X2 = 11.25, P = 0.012; X2 = 18.12, P = 0.002).The VEGF positive expression rates in infiltrating-type cases (75.4%), stageⅢ-Ⅳ(88.5%), vessel invasion (82.9%), lymphatic metastasis (84.3%) and distant metastasis (95.2%) were significantly higher than those of the expanding-type (X2 = 9.61, P = 0.021), stage I-II (X2=16.66, P = 0.001), non-vessel invasion (X2= 29.38, P = 0.001), non-lymphatic metastasis (X2 = 18.68, P = 0.005), and non-distant metastasis (X2= 22.72, P = 0.007; X2 = 21.62, P = 0.004). The mean MVD in the specimens positive for the uPA mRNA, uPAR mRNA and VEGF protein was markedly higher than those with negative expression groups. Moreover, a positive relation between MVD and uPA mRNA (rs = 0.199, P = 0.042), uPAR mRNA (rs = 0.278, P = 0.035), and VEGF (rs = 0.398, P = 0.048) expressions was observed. The mean survival time in cases with positive uPA mRNA, uPAR mRNA and VEGF protein expression or MVD value≥54.9 was significantly shorter than those in cases with negative expression or MVD value < 54.9. CONCLUSION: uPA and uPAR expressions are correlated with enhanced VEGF-induced tumor angiogenesis and may play a role in invasion and nodal metastasis of gastric carcinoma, thereby serving as prognostic markers of gastric cancer.     

TGF beta1 expression and angiogenesis in colorectal cancer tissue

AIM: Transforming growth factor TGF beta1 is involved in a variety of important cellular functions,including cell growth and differentiation, angiogenesis, immune function and extracellular matrix formation. However, the role of TGF beta(1) as an angiogenic factor in colorectal cancer is still unclear. We investigate the relationship between transforming growth factor beta(1) and angiogenesis by analyzing the expression of transforming growth factor TGF beta(1) in colorectal cancer, as well as its association with VEGF and MVD. METHODS: The expression of TGF beta(1),VEGF, as well as MVD were detected in 98 colorectal cancer by immunohistochemical staining. The relationship between the TGF beta(1) expression and VEGF expression,MVD was evaluated. To evaluate the effect of TGF beta(1) on the angiogenesis of colorectal cancers. RESULTS: Among 98 cases of colorectal cancer,37 were positive for TGF beta(1) 37.8% 36 for VEGF 36.7% respectively. The microvessel counts ranged from 19 to 139.8, with a mean of 48.7(standard deviation,21.8).The expression of TGF beta(1) was correlated significantly with the depth of invasion, stage of disease, lymph node metastasis, VEGF expression and MVD. Patients in T3-T4, stage III-IV and with lymph node metastasis had much higher expression of TGF beta(1) than patients in T1-T2, stage I-II and without lymph node metastasis (P<0.05). The positive expression rate of VEGF(58.3%) in the TGF-beta(1) positive group is higher than that in the TGF-beta(1) negative group(41.7%, P<0.05). Also, the microvessel count (54+/-18) in TGF-beta(1) positive group is significantly higher than that in TGF-beta(1) negative group(46+/-15, P<0.05). The microvessel count in tumors with both TGF-beta(1) and VEGF positive were the highest (58+/-20 36-140, P<0.05).Whereas that in tumors with both TGF-beta(1) and VEGF negative were the lowest (38+/-16, 19-60, P<0.05). CONCLUSION: TGF beta(1) might be associated with tumor progression by modulating the angiogenesis in colorectal cancer and TGF beta(1) may be used as a possible biomarker.     

Immunohistochemical expression of vascular endothelial growth factor (VEGF) does not correlate with microvessel density in renal cell carcinoma

The aim of present study was to investigate the relationship between the immunohistochemical expression of vascular endothelial growth factor (VEGF) and microvessel density (MVD) assessed by CD31 and endoglin (CD105) in renal cell carcinoma (RCC). Specimens from 45 cases of RCC. were formalin-fixed, paraffin embedded, and sections were stained with H&E. Additional sections from each case were stained for VEGF, CD31, CD105, and alpha smooth muscle cell actin (SMA). VEGF immunohistochemical expression was estimated as negative (0), weak positive (+1), moderate positive (+2), and intense positive (+3). Microvessel density (MVD) was estimated on 5 hot spots (x400) from each case, and the arithmetic media was the final result. MVD was separately calculated on slides stained with CD31 and CD105. The rate between mature and immature blood vessels was calculated on slides stained with CD31/CD105/SMA. Statistic analysis was performed with SPSS10.0. The immunoreaction for VEGF was positive in epithelial cells of the renal tubules, and occasionally, in endothelial cells. In RCC, tumor cells were positive in 34 from 45 cases (75.5%). 11 cases were negative, 14 were slightly positive (+1), 13 moderate (+2), and 7 intense (+3). No relationship was found between the expression of VEGF and pathological form and nuclear grade, excepting for the chromophilic variant (3 cases, all positive). CD31 was positive in all cases, and CD105 in 39 cases. The mean values of MVD on slides stained for CD31 and CD105 were: 31.68 (range 9.8-60.2)/20.66 (range 4.2-52.8). The rate CD31/SMA positive blood vessels was 1/0.62. VEGF was expressed in 75.5% of 45 cases with RCC, and the mean value of MVD CD31/CD105 was 31.68/20.66. The immunohistochemical expression of VEGF does not correlate with MVD performed on slides stained for both CD31 and endoglin. The majority of blood vessels in the tumor area are of mature type, with perivascular cells positive for SMA.     

Correlation of integrin β3 mRNA and vascular endothelial growth factor protein expression profiles with the clinicopathological features and prognosis of gastric carcinoma

AIM: To investigate integrin 133 mRNA and vascular endothelial growth factor (VEGF) protein expression in gastric carcinoma, and its correlation with microvascular density, growth-pattern, invasion, metastasis and prognosis. METHODS: In situ hybridization(ISH) of integrin β3 mRNA and immunohistochemistry of VEGF and CD34 protein were performed on samples from 118 patients with gastric cancer. RESULTS: The positive rate of integrin 133 mRNA in non- tumor gastric mucosa (20%) was significantly lower than that of the gastric cancer tissue (52.5%, x2 = 10.20, P ＜ 0.01). In patients of infiltrating type, stage T3-T4, vessel invasion, lymphatic metastasis, hepatic or peritoneal metastasis, the positive expression rates of integrin β3 mRNA were significantly higher than those in patients of expanding type (P ＜ 0.01), stage T1-T2 (P ＜ 0.01), non-vessel invasion (P ＜ 0.01), without lymphatic metastasis (P ＜ 0.01), without hepatic and peritoneal metastasis (P ＜ 0.01), respectively. In patients of infiltrating type, stage T3-T4, vessel invasion, lymphatic metastasis, hepatic or peritoneal metastasis, the positive expression rates of VEGF protein were significantly higher than those in patients of expanding type (P ＜ 0.01), stage T1-T2 (P ＜ 0.01), non-vessel invasion (P ＜ 0.01), without lymphatic metastasis (P ＜ 0.01), without hepatic and peritoneal metastasis (P ＜ 0.01), respectively. In patients of infiltrating type, stage T3-T4, vessel invasion, lymphatic metastasis, hepatic or peritoneal metastasis, the mean MVD were significantly higher than those in patients of expanding type (P ＜ 0.01), stage T1-T2 (P ＜ 0.01), non-vessel invasion (P ＜ 0.01), without lymphatic metastasis (P ＜ 0.01), without hepatic and peritoneal metastasis (P ＜ 0.01), respectively. It was found that the positive expression rate of integrin β3 mRNA was positively related to that of VEGF protein (P ＜ 0.01) and MVD (P ＜ 0.05), meanwhile the positive expression rate of VEGF protein was positively related to NVD (P ＜ 0.05). The mean survival period in patients with positive expression of integrin β3 mRNA and VEGF, and MVD ≥ 54.9/mm2 was significantly shorter than that in patients with negative expression of integrin β3 mRNA (P ＜ 0.05) and VEGF (P ＜ 0.01), and MVD ＜ 54.9/mm2 (P ＜ 0.01). Five-year survival rate in patients with positive expression of integrin β3 mRNA and VEGF, and MVD ≥ 54.9/mm2 was significantly lower than those with negative expression of integrin β3 mRNA (P ＜ 0.05), VEGF (P ＜ 0.05), and NVD ＜ 54.9/mm2 (P ＜ 0.01). CONCLUSION: Integrin β3 and VEGF expression can synergistically enhance tumor angiogenesis, and may play a crucial role in invasion and metastasis of gastric carcinoma. Therefore, they may be prognostic biomarkers and novel molecular therapeutic targets.     

利用组织芯片技术研究PTEN和VEGF在胃癌中的表达及意义

目的研究PTEN和血管内皮生长因子（vascular endothelial growth factor，VEGF）在胃癌中的表达及临床意义。方法应用组织微阵列仪制作97孔胃癌组织芯片（tissue microarray）。用免疫组织化学S—P法检测PTEN、VEGF在72例胃癌和25例正常胃黏膜中的表达。结果胃癌组织中PTEN蛋白阳性表达率显著低于正常胃黏膜（45．8％ VS 100％，P〈0．01）；VEGF的阳性表达率显著高于正常胃黏膜（75％VSl2％，P〈0．01），PTEN在胃癌中的表达与VEGF呈负相关（P〈0．01）。PTEN、VEGF的表达在中高分化腺癌分别为68．8％、62．5％（P〉0．05），在低分化及未分化腺癌分别为27．5％、85．0％（P〈0．05）；伴淋巴结转移者分别为31．6％、86．9％（P〈0．05），无淋巴结转移者分别为61．8％、61。8％（P〉0．05）；临床病理分期Ⅰ＋Ⅱ期分别为57．1％、61．9％（P〉0．05），Ⅲ＋Ⅳ期分别为30．0％、93．3％（P〈0．05）；与性别、年龄、肿瘤大小和组织分型无显著差异（P〉0．05）。结论PTEN失活或蛋白表达降低、VEGF的高表达与胃癌临床病理特征和生物学行为有密切关系。PTEN在低分化或未分化以及伴淋巴结转移和临床Ⅲ＋Ⅳ期胃癌中的表达与VEGF呈负相关。联合检测PTEN、VEGF对胃癌的恶性程度及预后判断具有一定的临床参考意义。应用组织芯片大规模高效检测临床组织样本是可行的，具有快速、准确、方便经济的特点。     

血管内皮细胞生长因子的表达及微血管密度与胆囊癌浸润、转移及预后的相关研究

探讨血管内皮细胞生长因子 (vascularendothelialgrowthfactor,VEGF)及微血管密度 (Microvesseldensity ,MVD)在胆囊癌发生发展中的作用及与胆囊癌浸润、转移及预后的关系。应用S -P免疫组化技术对 3 1例经手术切除的原发性胆囊癌及 10例经手术切除的慢性胆囊炎标本进行VEGF蛋白和微血管密度检测。 3 1例胆囊癌组织中癌旁VEGF表达及MVD值均明显高于癌中央及正常组织 ,三者差异具有显著性 (P <0 0 1) ;VEGF表达与MVD具有相关性 ,VEGF阳性者MVD值显著高于阴性者 (P <0 0 1) ;VEGF表达和MVD与胆囊癌分化程度、浸润转移、Nevin分期密切相关 (P <0 0 1) ;VEGF阳性者及高MVD者预后较阴性者差 ;Cox比例危险模型多因素分析表明 :VEGF对胆囊癌是一个独立的预后因子。VEGF的表达及MVD在胆囊癌的发生和浸润转移过程中发挥重要的作用 ,VEGF和MVD可作为反映胆囊癌生物学行为的指标     

Correlative studies on bFGF mRNA and MMP-9 mRNA expressions with microvascular density, progression, and prognosis of gastric carcinomas

AIM: To investigate the mRNA expressions of bFGF and MMP-9 in gastric carcinomas so as to reveal their correlations with tumor microvascular density (MVD),invasion, metastasis, and prognosis. METHODS: In situ hybridization and immunohistochemical techniques were used to detect the expressions of bFGFmRNA and MMP-9mRNA and the proteins of CD34 in 105 specimens of gastric carcinomas. RESULTS: In situ hybridization study showed that positive rates of bFGF mRNA and MMP-9mRNA expressions were 60.95% and 59.19%; the mean MVD was 46.09±11.52 and 43.75±13.41, respectively in piece/0.72 mm2 in tumors with bFGFmRNA and MMP-9mRNA positive expressions, which were significantly higher than those with negative expression (29.41±12.47; 33.45±13.92 piece/0.72 mm2, respectively). The positive expression rates of bFGFmRNA and MMP-9mRNA were correlated to the tumor invasion depth (rs= 0.211, P= 0.031; rs= 0.335, P= 0.001), growing pattern (rs= 0.324, P= 0.001; rs= 0.267, P= 0.006), vessel invasion (rs= 0.579, P= 0.001; rs = 0.209, P= 0.032), lymph node metastasis (rs= 0.405, P= 0.001; rs= 0.343, P= 0.001) and distant metastasis (rs= 0.474, P= 0.001; rs = 0.468, P = 0.001), but not correlated to tumor type (rs=0.134,P=0.173;rs=0.103,P=0.145) and differentiations (rs=0.096,P= 0.332;rs=0.102,P=0.298). The mean MVD was much higher in the tumors with infiltrating growth at stage T3-T4, with vessel invasion, lymph node metastasis and distant metastasis than those with expanding growth type (t = 10.105, P= 0.001) at stage T1-T2 (t=5.961,P=0.001),with non-vessel invasion (t=7.394,P=0.001),non-lymph node metastasis (t = 3.819, P= 0.01) and non-distant metastasis (t = 10.578, P= 0.001). Positive correlation was observed between MVD and the expressions of bFGFmRNA and MMP-9mRNA (t = 3.207, P=0.002; t = 7.035, P= 0.001, respectively). The mean survival time and 5-year survival rate were lower in cases with MVD over 39.5 and the positive expressions of bFGFmRNA and MMP-9mRNA than those with MVD less than 39.5 and the negative expressions of bFGFmRNA and MMP-9mRNA. CONCLUSION: bFGF and MMP-9 promote the angiogenesis of the gastric cancers. Detection of the expressions of bFGF and MMP-9 can serve as a useful index to determine the angiogenesis, invasion, metastasis, and prognosis of gastric cancers.     

Overexpression of EphA2, MMP‐9, and MVD‐CD34 in hepatocellular carcinoma: Implications for tumor progression and prognosis

Aim: To investigate the expression of erythropoietin‐producing hepatocellular (Eph)A2 receptor, matrix metalloproteinase (MMP)‐9, and angiogenesis in hepatocellular carcinoma (HCC), in order to reveal their expression correlations with tumor invasion, metastasis, and prognosis. Methods: From January 2000 to June 2003, 129 specimens of resected tumors from the patients with HCC were obtained. Corresponding pericarcinomatous liver tissues were also obtained and selected as a control group. Expressions of EphA2, MMP‐9, and CD34 were detected with immunohistochemical staining. Microvascular density (MVD) was calculated with counting of CD34‐positive vascular endothelial cells. Results: The expressions of EphA2, MMP‐9, and MVD in the HCC tissues were significantly higher than those in the pericarcinomatous liver tissues (P < 0.01). Statistical analysis showed there were significant correlations between the expressions of EphA2, MMP‐9 and MVD in some classicclinicopathological parameters (i.e. tumor nodule, vein invasion, tumor, node, metastasis stages, extrahepatic metastasis; P < 0.05). The correlation between EphA2 and MMP‐9 expression was positive (r = 0.625, P = 0.011). Tumor MVD was closely associated with EphA2 (r = 0.281, P = 0.01) and MMP‐9 (r = 0.319, P < 0.01) expressions. In particular, EphA2, MMP‐9, and MVD expressions levels were found to be independent prognostic factors after HCC resection. Conclusions: Overexpressions of EphA2 and MMP‐9 relate to tumor progression, metastasis, and prognosis in HCC. The present study suggests that EphA2 is associated with key mediators of angiogenesis and invasion.     

微血管密度与CD105、CD31在胃癌中的表达及意义

目的探讨胃癌组织中微血管密度（microvessel density,MVD）与CD105、CD31的表达及生物学行为的关系。方法取61例术前未作放、化疗及免疫治疗胃癌病人的外科手术切除标本,采用免疫组织化学技术S-P法检测CD105、CD31的表达,并定量计数胃癌组织微血管。结果胃癌组织中CD105及CD31标记的MVD在与淋巴结转移、肿瘤浸润深度均有显著性差异（P〈0．05）;Kaplan．Meier法分析显示微血管计数CD105〈14个,CD31〈34个（200×）的胃癌病人预后较好,而微血管计数CD105≥14个,CD31≥34个（200×）预后较差。结论胃癌MVD与其生物学行为有关,提示肿瘤血管生成分析和微血管密度计数是评价胃癌预后的一项重要参考指标。