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.     

HPSE、MMP-9和E-cad联合表达对中老年非小细胞肺癌侵袭、转移及其预后的影响

目的研究中老年非小细胞肺癌(NSCLC)中HPSE、MMP-9、E-cad mRNA和蛋白的表达,探讨其与肿瘤侵袭性、淋巴结转移及预后的关系。方法应用实时荧光定量PCR和免疫组织化学方法检测NSCLC组织中HPSE、MMP-9、E-cad mRNA和蛋白表达。同时,对随访资料进行生存分析,绘制生存率曲线,探讨三者对NSCLC患者预后的影响。另外,通过细胞黏附能力实验测定硫酸多糖对NSCLC A549细胞黏附能力的影响。结果HPSE、MMP-9 mRNA和微血管密度(MVD)值在不同肺组织中的表达有显著性差异(均P<0.05)。HPSE、MMP-9、E-cad的表达在不同年龄、性别、组织学类型及分化程度间无显著性差异(均P>0.05),而在淋巴结转移与否组间有显著性差异(P<0.05)。患者术后生存期与HPSE、MMP-9及E-cad的表达强度显著相关(均P<0.05)。生存率曲线显示HPSE和MMP-9蛋白高表达者生存时间均短于低/无表达者,E-cad蛋白则与之相反(P=0.027,0.018,0.030,n=65)。HPSE和MMP-9的表达与MVD值均呈正相关(r=0.48;r=0.54)。细胞黏附能力实验显示实验组NSCLCA549细胞黏附率明显比对照组低。结论HPSE、MMP-9、E-cad表达可作为预测中老年NSCLC侵袭、淋巴结转移和预后的良好指标;三者共同促进了NSCLC微血管形成和肿瘤细胞转移;HPSE抑制剂硫酸多糖能够有效抑制NSCLCA549细胞的黏附能力。     

胃癌组织中PTEN,MMP-9和Caspase-3表达的关系

目的:研究PTEN,MMP-9和Caspase-3在胃癌及正常胃组织中的表达,探讨他们在胃癌的发生、发展、浸润和转移中的作用．方法:选择临床病理资料齐全的胃癌蜡块标本54例,另取正常胃黏膜标本15例作对照．采用SP免疫组化方法检测PTEN,MMP-9和 Caspase-3在其中的表达．结果:胃癌中PTEN低表达(28/54,51．9％),且肿瘤浸润深(P=0．004)、有淋巴(P=0．003) 和远隔转移(P=0．01 5)、临床分期高(P= 0．001)、病理分化低(P=0．008)时降低．胃癌中MMP-9高表达(41／54,75．9％),且肿瘤浸润深(P=0．040)、有淋巴转移(P=0．025)、临床分期高(P=0．039)、病理分化低(P=0．009)时增高．胃癌中Caspase-3低表达(12／54,22．2％), 且有淋巴转移(P=0．045)、临床分期高(P= 0．015)、病理分化低(P=0．035)时降低．胃癌中PTEN与MMP-9(r=-0．543,P=0．001), Caspase-3与MMP-9的表达负相关(r=0．741, P=0.001),PTEN与Caspase-3的表达正相关(r =0．515,P=0．001)．结论:胃癌中PTEN,Caspase-3低表达,MMP-9 高表达;PTEN、MMP-9和Caspase-3可作为胃癌诊断和预后判断的指标．     

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）,     

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&#177;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&#177;14.96,67.09&#177;18.29,respectively)were higher than those in stageⅠ,Ⅱ(t\2.378,4.015,2.503and2.450,P&lt;0.05,P&lt;0.001,P&lt;0.001,P&lt;0.05,respectively),In37gastric carcinoma specimens with lymph node metastasis,MVD(68.69&#177;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&lt;0.01,P&lt;0.05,P&lt;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&#177;18.02,58.39&#177;17.73,repectively)were higher than those in the negative iNOS and VEGF specimens(X^2=6.3587and 6.1574,P&lt;0.05,P&lt;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&lt;0.05,P&lt;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.     

COX-2 expression in gastric cancer and its relationship with angiogenesis using tissue microarray

AIM： To explore the expression and clinicopathological significance of cyclooxygenase-2 （COX-2） and microvessel density （MVD） in gastric carcinogenesis, and to investigate their roles in the invasion and the relationship between biological behaviors and prognosis of gastric cancer.
METHODS： Using Envision immunohistochemistry, COX-2 and CD34 expressions in gastric cancer tissue array were examined. MVD was counted and the relationship between the biological behaviors and prognosis was analyzed.
RESULTS： The expression of COX-2 in gastric cancer tissue was significantly higher than that in normal mucosa （χ^2 = 12.191, P 〈 0.05）. The over-expression of COX-2 in gastric cancer was obviously related to metastasis and depth of invasion （χ^2 = 6.315, P 〈 0.05）, but not related to the histological type and Borrmann type （χ^2 = 5.391 and χ^2= 2.228, respectively）. Moreover, MVD in gastric cancer tissues was significantly higher than that in the normal mucosa （65.49 ± 20.64 vs 36.21 ± 18.47, t/F = 7.53, P 〈 0. 05）. MVD was related to the histologic type and metastasis （t/F= 3.68 and t/F = 4.214, respectively, P 〈 0. 05）, but not related to the depth of invasion and Borrmann type （t/F = 0.583 and t/F = 0.459, respectively）. MVD in COX-2-positive tissues was markedly higher compared to COX-2-negative tissues, indicating a positive correlation between COX-2 expression and MVD （t = 13.12, P 〈 0. 05）.
CONCLUSION： Tissue microarray （TMA） is a powerful tool for rapid identification of the molecular alterations in gastric cancer. COX-2 expression, via inducingangiogenesis, may play an important role in gastric carcinogenesis. It could be served as a determinant factor for clinical prognosis and curative effect.     

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.     

Expressions of inducible nitric oxide synthase and matrix metalloproteinase-9 and their effects on angiogenesis and progression of hepatocellular carcinoma

AIM: To determine the expressions of inducible nitric oxide synthase (iNOS) and matrix metalloproteinase-9 (MMP-9) in hepatocellular carcinoma (HCC) and to investigate the relationship between iNOS and MMP-9 expression and their effects on angiogenesis and progression of HCC. METHODS: In this study, we examined iNOS, MMP-9, and CD34 expression in specimens surgically removed from 32 HCC patients and 7 normal liver tissues by immunohistochemical staining. Meanwhile, microvessel density (MVD) was determined as a marker of angiogenesis by counting CD34-positive cells. RESULTS: The positive rates of iNOS and MMP-9 expression were 71.88% (23/32) and 78.13% (25/32) in HCC. MMP-9 expression was significantly correlated with tumor size, capsule status, TNM stage, and risk of HCC recurrence (P= 0.032, P= 0.033, P= 0.007, and P= 0.001, respectively). There was also a significant relationship between iNOS expression and capsule status and risk of HCC recurrence (P= 0.049 and P= 0.004, respectively), but no correlation between iNOS expression and tumor size and TNM stage. There was a positive association between MVD and TNM stage and risk of HCC recurrence (P= 0.037 and P= 0.000, respectively). The count of MVD was significantly different in different iNOS and MMP-9 immunoreactivity groups (F= 17.713 and 17.097, P= 0.000 and P= 0.000, respectively). The examination of Spearman's rank correlation coefficient showed that there was a significant positive correlation between MVD and iNOS, MMP-9 immunoreactivity (r= 0.754 and 0.751, P= 0.000 and A=0.000, respectively). There was also a significant association between MMP-9 and iNOS expression in HCC (P= 0.010). CONCLUSION: Nitric oxide (NO) produced by iNOS could modulate MMP-9 production and therefore contribute to tumor cell angiogenesis and invasion and metastasis in HCC. The strong expression of iNOS and MMP-9 in HCC may be helpful in evaluating the recurrence of HCC, predicting poor prognosis. For patients with strong expression of MMP-9 and iNOS, the optimal treatment scheme needs to be selected.     

胃癌组织中KISS-1和MMP-9的表达及其意义

目的探讨肿瘤转移抑制基因K ISS-1及基质金属蛋白酶9（MMP-9）与胃癌侵袭、转移的关系,为研究胃癌的转移机制及治疗提供理论基础。方法采用逆转录聚合酶链反应（RT-PCR）检测36例胃癌组织及36例正常胃组织中K ISS-1 mRNA及MMP-9 mRNA的表达情况,分析其与胃癌患者各临床病理指标的关系及二者的相关性。结果 K ISS-1 mRNA在胃癌组织中的阳性表达率及表达水平均低于正常胃组织（P均〈0.01）,并且其低表达与淋巴结转移密切相关（P〈0.05）;MMP-9 mRNA在胃癌组织中的阳性表达率及表达水平均高于正常胃组织（P均〈0.05）,MMP-9 mRNA的高表达与癌的浸润深度和淋巴结转移密切相关（P均〈0.05）;K ISS-1与MMP-9表达呈负相关（P〈0.05）。结论 K ISS-1表达缺失和MMP-9过表达可能与胃癌的侵袭相关。     

Relationship between matrix metalloproteinase-2 mRNA expression and clinicopathological and urokinase-type plasminogen activator system parameters and prognosis in human gastric cancer

AIM: To investigate the relationship between matrix metalloproteinase-2 (MMP-2) mRNA expression and clinicopathologic and urokinase-type plasminogen activator (uPA) system parameter and prognosis in human gastric cancer. METHODS: Expression of MMP-2 mRNA, uPA, and uPA-R mRNA in tumor tissues and ≥5 cm adjacent normal tissues from 67 cases of gastric cancer was studied using RT-PCR and Northern blot respectively.Survival analyses were done using the Kaplan-Meier method. RESULTS: The expression rates of MMP-2 mRNA,uPA and uPA-R mRNA in tumor tissues (31%,41%,and 51%, respectively) were significantly higher than those in ≥5 cm adjacent tissues (19%, 11%, and 9%; X2=4.59,43.58, and 53.24 respectively, P<0.05,0.0001,and 0.0001, respectively). Expression of MMP-2 mRNA was significantly correlated with lymph node metastasis (metastasis: 61.9%, no metastasis: 39.1%, X2= 7.61, P<0.05),Lauren's classification of diffuse/mixed types:54.2%,intestinal type: 26.3%,X2 = 4.25, P<0.05, expression of uPA and uPA-R mRNA (uPA+: 55.1%, uPA-: 22.2% and uPA-R+: 54.9%, uPA-R-: 18.8%, X2=5.72 and 6.40 respectively, P<0.05).Kaplan-Meier survival analysis of MMP-2 mRNA expression did not show significant difference in all 67 cases, but revealed an association of the expression of MMP-2 mRNA, uPA, and uPA-R mRNA with worse prognosis (P= 0.0083, 0.0160, and 0.0094, respectively). CONCLUSION: MMP-2 may play an important role in the development of invasion and metastasis of gastric cancer.     

Expression and significance of CD44s, CD44v6, and nm23 mRNA in human cancer

AIM: To investigate the relationship between the expression levels of nm23 mRNA, CD44s, and CD44v6,and oncogenesis, development and metastasis of human gastric adenocarcinoma, colorectal adenocarcinoma,intraductal carcinoma of breast, and lung cancer.METHODS: Using tissue microarray by immuhistochemical (IHC) staining and in situ hybri-dization (ISH), we examined the expression levels of nm23mRNA, CD44s, and CD44v6 in 62 specimens of human gastric adenocarcinoma and 62 specimens of colorectal adenocarcinoma; the expression of CD44s and CD44v6in 120 specimens of intraductal carcinoma of breast and 20 specimens of normal breast tissue; the expression of nm23 mRNA in 72 specimens of human lung cancer and 23 specimens of normal tissue adjacent to cancer.RESULTS: The expression of nm23 mRNA in the tissues of gastric and colorectal adenocarcinoma was not significantly different from that in the normal tissues adjacent to cancer (P＞0.05), and was not associated with the invasion of tumor and the pathology grade of adenocarcinoma (P＞0.05). However, the expression of nm23 mRNA was correlated negatively to the lymph node metastasis of gastric and colorectal adenocarcinoma (r = -0.49, P＜0.01; r = -4.93, P＜0.01). The expression of CD44s in the tissues of gastric and colorectal adenocarcinoma was significantly different from that in the normal tissues adjacent to cancer (P＜0.05;P＜0.01). CD44v6 was expressed in the tissues of gastric and colorectal adenocarcinoma only, the expression of CD44v6 was significantly associated with the lymph node metastasis, invasion and pathological grade of the tumor (r = 0.47, P＜0.01; r = 5.04, P＜0.01). CD44sand CD44v6 were expressed in intraductal carcinoma of breast, the expression of CD44s and CD44v6 was significantly associated with lymph node metastases and invasion (P＜0.01). However, neither of them was expressed in the normal breast tissue. In addition, the expression of CD44v6 was closely related to the degree of cell differentiation of intraductal carcinoma of breast (x2= 5.68, P＜0.05). The expressional level of nm23mRNA was closely related to the degree of cell differentiation (P＜0.05) and lymph node metastasis (P＜0.01), but the expression of nm23 gene was not related to sex, age, and type of histological classification (P＞0.05).CONCLUSION: Patients with overexpression of CD44s and CD44v6 and low expression of nm23 mRNA have a higher lymph node metastatic rate and invasion. In addition, overexpression of CD44v6 is closely related to the degree of cell differentiation. Detection of the three genes is able to provide a reliable index to evaluate the invasion and metastasis of tumor cells.     

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.     

Imbalance between expression of matrix metalloproteinase-9 and tissue inhibitor of metalloproteinase-1 in invasiveness and metastasis of human gastric carcinoma

AIM: The expressive balance between matrix metalloproteinase-9 (MMP-9) and its tissue inhibitor of metalloproteinase-1 (TIMP-1) plays a critical role in maintaining the degradation and synthesis of extracellular matrix. Loss of such balance is associated with invasion and metastasis of tumors. This study aimed to determine the expression of MMP-9 and TIMP-1 in gastric carcinoma, and the association of the expressive imbalance between MMP9 and TIMP-1 with the invasion and metastasis and prognosis of gastric carcinoma.METHODS: We used immunohistochemistry to determine the expressions of MMP-9, TTMP-1 and proliferating cell nuclear antigen Ki-67 in the gastric specimens taken from 256 patients with primary gastric carcinoma. The patients were followed-up for up to 96 months.RESULTS: No association between the expression of MMP9 and TIMP-1 and patients' sex and age, tumor size and location of gastric carcinoma was observed. The incidence of the positive expression of MMP-9 in cases with tumors invasion to muscularis propria and visceral peritoneum (70.13% and 69.09%, respectively) was significantly higher than that in cases with tumor invasion only to lamina propria or submucosa (42.50 %, P=0.0162). The positive correlation between MMP-9 expression and the depth of tumor invasion was observed (Pearson correlation coefficient=0.2129,P=0.016). Along with the increase of the metastatic station of lymph nodes, the incidence of the MMP-9 expression was increased by degrees; a positive correlation between them was observed (Pearson correlation coefficient=0.2910,P=0.0001). There was also a significant correlation between MMP-9 expression and the TNM stage in gastric carcinoma (Pearson correlation coefficient=0.3027, P＜0.0001). The incidence of MMP-9 expression in stage Ⅱ and Ⅲ/Ⅳ (75.00%and 76.15%, respectively) was significantly higher than those in stage Ⅰ (46.15 %, P＜0.0001). A negative correlation between TIMP-1 immunoreactivity and the depth of invasion,status of lymph node metastasis and TNM stage was observed (Pearson correlation coefficient =-0.1688, -0.3556and -0.3004, P=0.023, ＜0.0001 and ＜0.0001, respectively).Four types of co-expression of MMP-9 and TIMP-1 were observed; i.e. MMP-9 positive but T IMP-1 negative (n=115),both positive (n=52), both negative (n=62) and MMP-9negative but TIMP-1 positive (n=27). The frequency of serosal invasiveness was significant higher in patients with MMP-9 but without TIMP-1 expression than those with other types of the co-expression (P=0.0303). The incidence of lymph node metastasis was highest in patients with MMP-9but without TIMP-1 expression, and lowest in those with TIMP-1 but without MMP-9 expression (P＜0.0001). The survival rate in patients with MMP-9 but without TIMP-1expression was lower than that in those with TIMP-1 but without MMP-9 expression (P=0.0014).CONCLUSION: Our results in gastric carcinoma demonstrated a significant positive association of MMP-9 over-expression with proliferation of tumor cells, the depth of invasiveness,lymph node metastasis and TNM stage, suggesting MMP-9can serve as a molecular marker of tumor invasion and metastasis. We also demonstrate a significant negative relationship of TIMP-1 expression with the depth of invasiveness and lymph node metastasis, which provide a new idea in the tumor biological and genetic treatment.The interaction between MMP-9 and TIMP-1 in the processes of tumor invasion and metastasis is that MMP-9 mainly promotes tumor invasion and metastasis and TIMP-1 inhibits functions of MMP-9. The imbalance between MMP-9 and TIMP-1 expression may suggest the occurrence of tumor invasion and metastasis, predict poor prognosis. For patients with imbalanced MMP-9 and TIMP-1 expression, the optimal treatment scheme needs to be selected.     

结肠癌COX-2mRNA表达与微血管密度关系的临床意义

目的通过观察结肠癌组织COX-2mRNA及CD34的表达，结合结肠癌组织中微血管密度（microvessel density，MVD）所见，探讨COX-2与肿瘤血管形成及病理特征的关系，为结肠癌生物治疗提供理论基础。方法 选择62例结肠癌、22例结肠腺瘤和22例正常结肠黏膜标本，采用原位分子杂交法检测COX-2mRNA并用MaxVisionTM快捷免疫组化法检测CD34表达，光镜下记数MVD。结果结肠癌、结肠腺瘤组织COX-2mRNA的阳性率明显高于正常黏膜，且有统计学意义（74．19％：36．36％，P=0．001；72．73％：36．36％，P=0．015）；结肠癌组平均MVD值高于腺瘤组和正常组，三组比较，有统计学意义（F=19．628，P=0．000）。在62例结肠癌组织中，高分化组COX-2mRNA阳性率高于低分化组（X^2=4．215、P=0．040）；进展期癌组的MVD高于早期癌组（t=3．079，P：0．003）；淋巴结有转移组MVD高于无转移组（t=3．180，P=0．002）；有血管侵犯组高于无血管侵犯组（t=2．093，P=0．041）；COX-2mRNA阳性组MVD高于阴性组，COX-2mRNA高表达组MVD高于低表达组，但差异均无统计学意义（P〉0．05）。结论MVD记数可作为判断肿瘤预后的有效评价指标，COX-2与肿瘤细胞的增殖和凋亡密切相关，与肿瘤血管生成无直接相关性。     

食管鳞癌中鸟氨酸脱羧酶mRNA、内皮抑素mRNA表达和血管生成相关性研究

目的 研究鸟氨酸脱羧酶(0DC)mRNA、内皮抑素mRNA和微血管密度(MVD)在食管鳞癌中的表达及相关关系，探讨0Dc在食管鳞癌血管生成中的可能作用，及其与肿瘤浸润和转移的关系。方法 用半定量RT-PcR法测定41例食管鳞癌患者痛组织(T)和相应癌旁组织(N)中的ODC mRNA和内皮抑素mRNA表达，求得各自T／N值；同时用免疫组化方法测定癌组织和癌旁正常组织MVD的T／N值。结果在41例患者中，39例ODC mRNA的T／N值＞1．0，占95．1％：40例MVD的T／N值＞1．0，占97．6％。36例内皮抑素mRNA的T／N值＞1．0，占87．8％。ODC mRNA、内皮抑素mRNA和MVD与食管鳞癌的肿瘤大小、浸润深度和淋巴结转移有关，ODC mRNA、内皮抑素mRNA表达与肿瘤分化程度有关。ODC mRNA的T／N值与MVD的T／N值呈正相关，与内皮抑素mRNA的T／N值呈负相关，内皮抑素mRNA的T／N值与MVD的T／N值呈负相关。结论 DOC与食管鳞癌血管生成和肿瘤浸润转移密切相关。DOC过表达可能是通过抑制内皮抑素而促进血管生成．从而促进食管鳞癌的浸润和转移。     

胃癌组织Maspin,uPA,MMP-7表达的意义

目的:观察胃癌及正常胃黏膜Maspin,uPA, MMP-7表达的意义．方法:应用免疫组化SP法检测胃管状腺癌30 例,胃印戒细胞癌30例,正常胃黏膜组织20例中Maspin,uPA,MMP-7的表达情况．结果:在胃管状腺癌中Maspin,uPA,MMP-7 阳性表达率分别为50％,70％和80％;胃印戒细胞癌中阳性表达率分别为46．7％,76．7％和 90％;正常胃黏膜组织中阳性表达率分别为 90％,35％和30％．Maspin的表达与浸润深度、淋巴结转移相关,而与肿块的大小和TNM分期无关．uPA和MMP-7的表达与浸润深度、淋巴结转移、TNM分期相关,而与肿块的大小无关．Maspin的表达与uPA和MMP-7的表达呈负相关(P=0．012,r=-0．322;P=0．008,r= -0．341);uPA的表达与MMP-7的表达呈正相关 (P=0．034,r=0．274)．结论:Maspin在胃癌中表达下调,uPA和 MMP-7在胃癌中过表达,他们在胃癌的浸润转移中起重要作用,可作为反应胃癌病理生物学行为的有效指标．     

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

AIM: To investigate the expression of PTEN/MMAC1/TEP1and 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 between PTEN 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 gastric 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 (γ = 0.512, P＜0.05). VEGF expression in PTEN-negative gastric 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. PTENrelated 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.