Expression of focal adhesion kinase and α5andβ1integrins in carcinonas and its clinical significance

AIM：To detect the expression pattern of FAK(focal adhesion kinase)and integrinα5andβ1 subunits in different kinds of cancerous tissues and to study their correlation with clinicopathological data including tumor type,grade and lymph node status.METHODS:Using an immunohistochemical technique,we examined the expression of FAK and integrin and subunits in cancerous and noncancerous tissues obtained from75patients with gastric carcinomas,21colorectal carcinomas,16 hepatocellular carcinomas,20uterocervical carcinomas,and20breast carcinomas.RESULTS:The staining of FAK was stronger in cancerous than in noncancerous areas,Enhanced expression of FAKwas detected in poor-differentiated carcinoma of the stomach and colorectum.Tumors with lymph node metastases had more FAKprotein than those without metastases.In addition.the deeper the extent of tumor infiltration,the higher the FAKexpression.The expression of integrinα5andβ1subunits was lower in cancerous areas than in noncancerous areas,but it was higher in well-differentiated cancerous tissues than in poor differentiated tissues.The relationship between the expression of integrinα5andβ1subunits and infiltration or metastasis was not significant.Cancerous tissues with stronger FAK expression(++or+++)also had a higher expression of integrinα5andβ1subunits in the tumor and its unaffected margins.CONCLUSION:FAKis a better marker for carcinogenesis and the progression of cancer than integrinα5orβ1subunit,and it may be not only a transformation-linked enzyme but also a progression-linked enzyme.     

Expression of focal adhesion kinase and α5 and β1 integrins in carcinomas and its clinical significance

AIM: To detect the expression pattern of FAK (focaladhesion kinase) and integrin α5 and β1 subunits indifferent kinds of cancerous tissues and to study theircorrelation with clinicopathological data includingtumor type, grade and lymph node status. METHODS:Using an immunohistochemical technique, weexamined the expression of FAK and integrin andsubunits in cancerous and noncancerous tissuesobtained from 75 patients with gastric carcinomas, 21colorectal carcinomas, 16 hepatocellular carcinomas,20 uterocervical carcinomas, and 20 breast carcinomas.RESULTS: The staining of FAK was stronger in cancerousthan in noncancerous areas. Enhanced expression ofFAKwas detected in poor-differentiated carcinoma ofthe stomach and colorectum. Tumors with lymph nodemetastases had more FAK protein than those withoutmetastases. In addition, the deeper the extent of tumorinfiltration, the higher the FAK expression. Theexpression of integrin α5 and β1 subunits was lower incancerous areas than in noncancerous areas, but it washigher in well-differentiated cancerous tissues than inpoor differentiated tissues. The relationship betweenthe expression of integrin α5 and β1 subunits andinfiltration or metastasis was not significant. Cancerous tissues with stronger FAK expression (++ or +++) alsohad a higher expression of integrin α5 and β1 subunitsin the tumor and its unaffected margins.CONCLUSION: FAK is a better marker for carcinogenesisand the progression of cancer than integrin α5 or β1subunit, and it may be not only a transformation-linkedenzyme but also a progression-linked enzyme.     

S-phase delay in human hepatocellular carcinoma cells induced by overexpression of integrin β1

AIM: To clarify the mechanisms of integrin overexpression in negatively regulalting the cell cyde control of hepatocellular carcinoma cells SMMC-7721.METHODS: The cell cycle pattern was determined by flow cytometry. The mRNA and protein expression levels were assayed by RT-PCR and Western blot, respectively. Stable transfection was performed by Upofectamine 2000 reagent,and cells were screened by G418.RESULTS: Overexpression of α5β1 or β1 integrin induced S-phase delay in SMMC-7721 cells, and this delay was possibly due to the accumulaltion of cydin-dependent kinase inhibitors (CKIs) p21^cip1 and p27^kip1. The decrease of protein kinase B (PKB) phosphorylation was present in this signaling pathway, but focal adhesion kinase (FAK) was not involved.When phosphorylation of PKB was solely blocked by wortmannin, p27^kip1 protein level was increased. Moreover,S-phase delay was recurred when attachment of the parental SMMC-7721 cells was inhibited by the preparation of poly-HEME, and this cell cycle pattern was similar to that of β1-7721 or α5β1-7721 cells.CONCLUSION: S-phase delay induced by overexpression of integdn 151 subunit is attributed to the decrease of PKB phosphorylation and subsequent increases of p21^cip1 and p27^kip1 proteins, and may be involved in the unoccupied α5β1 because of lack of its ligands.     

S-phase delay in human hepatocellular carcinoma cells induced by overexpression of integrin β1

AIM:To clarify the mechanisms of integrin overexpression in negatively regulating the cell cycle control of hepatocellular carcinoma cells SMMC-7721.METHODS: The cell cycle pattern was determined by flow cytometry. The mRNA and protein expression levels were assayed by RT-PCR and Western blot, respectively. Stable transfection was performed by Lipofectamine 2000 reagent,and cells were screened by G418.RESULTS: Overexpression of α5β1 or β1 integrin induced S-phase delay in SMMC-7721 cells, and this delay was possibly due to the accumulation of cyclin-dependent kinase inhibitors (CKIs) p21cip1 and p27kip1. The decrease of protein kinase B (PKB) phosphorylation was present in this signaling pathway, but focal adhesion kinase (FAK) was not involved.When phosphorylation of PKB was solely blocked by wortmannin, p27kip1 protein level was increased. Moreover,S-phase delay was recurred when attachment of the parental SMMC-7721 cells was inhibited by the preparation of polyHEME, and this cell cycle pattern was similar to that of β1-7721 or α5β1-7721 cells.CONCLUSION: S-phase delay induced by overexpression of integrin β1 subunit is attributed to the decrease of PKB phosphorylation and subsequent increases of p21cip1 and p27kip1 proteins, and may be involved in the unoccupied α5β1because of lack of its ligands.     

Promotion of Sema4D expression by tumor-associated macrophages: Significance in gastric carcinoma

AIM To study the role of semaphorin 4 D(Sema4 D) expression promoted by tumor-associated macrophages(TAMs) in gastric carcinoma cells and its clinical significance in the invasion and metastasis of gastric carcinoma.METHODS CD68 and Sema4 D expression was analyzed in gastric carcinoma and adjacent normal tissues from 290 patients using the immunohistochemical streptavidinperoxidase method, and their relationships with clinicopathological features were evaluated. Human M2 macrophages were induced in vitro and co-cultured in non-contact with gastric carcinoma SGC-7901 cells. Changes in the secretory Sema4 D level in the SGC-7901 cell supernatant were measured using an enzymelinked immunosorbent assay. The effects of TAMs on SGC-7901 cell invasion and migration were assessed with invasion and migration assays, respectively.RESULTS CD68 and Sema4 D protein expression was significantly higher in gastric carcinoma tissues than in adjacent normal tissues(71.7% vs 33.8% and 74.5% vs 42.8%, respectively; P 0.01). CD68 and Sema4 D protein expression was significantly associated with histological differentiation, TNM stage, and lymph node metastasis(P 0.05), and their expression levels were positively correlated with one another(r = 0.467, P 0.01). In the in vitro experiment, secretory Sema4 D protein expression was significantly increased in the supernatant of SGC-7901 cells co-cultured with TAMs compared with the blank control(1224.13 ± 29.43 vs 637.15 ± 33.84, P 0.01). Cell invasion and metastasis were enhanced in the Transwell invasion and migration assays(P 0.01).CONCLUSION TAMs promote the invasion and metastasis of gastric carcinoma cells possibly through upregulated secretory Sema4 D protein expression. Combined detection of TAM markers, CD68 and Sema4 D, in gastric carcinoma tissue shows potential to predict the trend of gastric carcinoma progression.     

Blocking effects of genistein on cell proliferation and possible mechanism in human gastric carcinoma

AIM: To study the blocking effects of genistein on cell proliferation cycle in human gastric carcinoma cells (SGC-7901) and the possible mechanism. METHODS: MTT assay was applied in the detection of the inhibitory effects of genistein on cell proliferation. Flow cytometry was used to analyze the cell cycle distribution. Immunocytochemical technique and Western blotting were performed to detect the protein expression of cyclin D_1, cyclin B_1 and p21~(waf1/cip1). RESULTS: Genistein significantly inhibited the growth and proliferation of human gastric carcinoma cells (SGC-7901). Seven days after treatment with different concentrations of genistein (2.5, 5.0, 10.0, 20.0 μg/mL), the growth inhibitory rates were 11.2%, 28.8%, 55.3%, 84.7% respectively and cell cycles were arrested at the G(2)/ M phase. Genistein decreased cyclin D_1 protein expression and enhanced cyclin B_1 and p21~(waf1/cip1) protein expression in a concentration-dependent manner. CONCLUSION: The growth and proliferation of SGC-7901 cells can be inhibited by genistein via blocking the cell cycle, with reduced expression of cyclin D_1 and enhanced expression of cyclin B_1 and p21~(waf1/cip1) protein in the concentration range of 0-20 μg/mL.     

Study of Sonic hedgehog signaling pathway related molecules in gastric carcinoma

AIM: To study the expression of Sonic hedgehog pathway-related molecules, Sonic hedgehog (Shh) and Glil in gastric carcinoma. METHODS: Expression of Shh in 56 gastric specimens including non-cancerous gastric tissues, gastric adenocarcinoma, gastric squamous cell carcinoma was detected by RT-PCR, in situ hybridization and immunohistochemistry. Expression of Glil was observed by in situ hybridization. RESULTS: The positive rate of Shh and Glil expression was 0.0%, 0.0% in non-cancerous gastric tissues while it was 66.7%, 57.8% respectively in gastric adenocarcinoma, and 100%, 100% respectively in gastric squamous cell carcinoma. There was a significant difference between the non-cancerous gastric tissues and gastric carcinoma (P<0.05). Elevated expression of Shh and Glil in gastric tubular adenocarcinoma was associated with poorly differentiated tumors while the expression was absent in gastric mucinous adenocarcinoma. CONCLUSION: The elevated expression of Shh and Glil in gastric adenocarcinoma and gastric squamous cell carcinoma shows the involvement of activated Shh signaling in the cellular proliferation of gastric carcinogenesis. It suggests Shh signaling gene may be a new and good target gene for gastric tumor diagnosis and therapy.     

Expression of E-selectin, integrinβ_1 and immunoglobulin superfamily member in human gastric carcinoma cells and its clinicopathologic significance

AIM: To study the expression levels of E- selectin, integrinβ1 and immunoglobulin supperfamily member-intercellular adhesion molecule-1 (ICAM-1) in human gastric carcinoma cells, and to explore the relationship between these three kinds of cell adhesion molecules and gastric carcinoma. METHODS: The serum contents of E-selectin, integrinβ1 and ICAM-1 were detected by enzyme-linked immuno-sorbent assay (ELISA), in 47 healthy individuals (control group) and in 57 patients with gastric carcinoma (gastric carcinoma group) respectively prior to operation and 7 d after operation. RESULTS: The serum E-selectin, ECAM-1 and integrinβ1 were found to be expressed in both control and gastric carcinoma groups. However, they were highly expressed in patients with gastric carcinoma patients before operation or with unresectable tumours. The expression levels of ICAM-1 and integrinβ1 were significantly higher in gastric carcinoma patients than in controls (P < 0.01). A comparison of the E-selectin levels between the two groups showed statistically insignificant differnce (P = 0.64). In addition, the expression levels were all decreased substantially in the postoperative patients subjected to radical resection of the tumours, indicating that the high level expressions of these compounds might be the important factor for predicting the prognosis of these patients. CONCLUSION: Serum E-selectin, ICAM-1 and integrin pi expression levels are probably related to the metastasis and relapse of gastric cancer.     

Relationship between cell adhesion molecules expression and the biological behavior of gastric carcinoma

AIM： To evaluate the relationship between the expression of cell adhesion molecules （CAMs） and the biological behavior of gastric carcinoma. METHODS： Expression of syndecan-1, E-cadherin and integrin β3 were evaluated by immunohistochemical study in a total of 118 gastric carcinomas and 20 nontumor gastric mucosas. RESULTS： The expressions of syndecan-1 and E-cadherin were significantly lower in gastric carcinoma compared to non-tumor gastric mucosa, and the low expression rates were positively correlated to the tumor invasion depth, vessel invasion, lymph node metastasis and distant metastasis （P 〈 0.01 in all cases）. However, the expression of integrin β3 was significantly higher in gastric carcinoma compared to non-tumor gastric mucosa, and the high expression rates were positively correlated to the tumor invasion depth, vessel invasion, lymph node metastasis and distant metastasis （P 〈 0.01 in all cases）. In addition, the three protein expressions were correlated to the tumor growth pattern （P 〈 0.01, P 〈 0.01, and P 〈 0.05 respectively）, but not correlated to tumor differentiation （P 〉 0.05, P 〉 0.05 and P 〉 0.05 respectively）. Positive correlation was observed between the expressions of syndecan-1 and E-cadherin, but they which were negatively correlated to the expression of integrin β3 （P 〈 0.01 in all cases）. Univariate analysis demonstrated that the mean survival time and 5-year survival rate were lower in the cases with low expressions of syndecan-1 and E-cadherin and high expression of integrin β3 （P 〈 0.01, in all cases）. COX multivariate analysis showed that the expression level of syndecan-1 could be an independent prognostic index of gastric carcinoma （P 〈 0.01）, whereas E-cadherin and integrin β3 could not be independent indexes （P 〉 0.05, P 〉 0.05 respectively）.CONCLUSION： The low expression of syndecan-1 and E-cadherin and the high expression of integrin β3 are significantly correlated with the invasion and metastasis o     

β-catenin up-regulates the expression of cyclinD1, c-myc and MMP-7 in human pancreatic cancer: Relationships with carcinogenesis and metastasis

AIM: To investigate whether abnormal expression of β-catenin in conjunction with overexpression of cyclinD1, c-myc and matrix metalloproteinase-7 (MMP-7) correlated with the carcinogenesis, metastasis and prognosis of pancreatic cancer, and to analyze the relationship of β-catenin expression with cyclinDl, c-myc and MMP-7 expression. METHODS: Using immunohistochemistry,we examined the expression of β-catenin, cyclinD1,c-myc and MMP-7 in 47 pancreatic adenocarcinoma tissues, 12 pancreatic intraepithelial neoplasia (PanIN) and 10 normal pancreases, respectively. Proliferation cell nuclear antigen was also tested as the index of proliferative activity of pancreatic cancer cells. RESULTS: In 10 cases of normal pancreatic tissues, epithelial cells showed equally strong membranous expression of β-catenin protein at the cell-cell boundaries, but the expression of cyclinDl, c-myc and MMP-7 was negative. The expression of β-catenin, cyclinD1, c-myc and MMP-7 in PanIN and pancreatic adenocarcinoma tissues had no significant difference [6/12 and 32/47 (68.1%), 6/12 and 35/47 (74.5%), 5/12 and 33/47 (70.2%), 7/12 and 30/47 (63.8%), respectively]. The abnormal expression of β-catenin was significantly correlated to metastasis and one-year survival rate of pancreatic cancer, but had no relation with size, differentiation and cell proliferation. The expression of cyclinD1 was correlated with cell proliferation and extent of differentiation, but not with size, metastasis and one-year survival rate of the pancreatic cancer. The expression of c-myc was not correlated with size, extent of differentiation, metastasis and 1-year survival rate, but closely with cell proliferation of pancreatic cancer. The overexpression of MMP-7 was significantly associated with metastasis and 1-year survival rate of pancreatic cancer,but not with size, extent of differentiation and cell proliferation.There was a highly significant positive association between abnormal expression of β-catenin and overexpression of cyclinD1, c-myc and MMP-7 not only in PanIN (r= 1.000, 0.845, 0.845), but also in pancreatic cancer (r= 0.437, 0.452, 0.435). CONCLUSION: The abnormal expression of β-catenin plays a key role in the carcinogenesis and progression of human pancreatic carcinoma by up-regulating the expression of cyclinDl, c-myc and MMP-7, resulting in the degradation of extracellular matrix and uncontrolled cell proliferation and differentiation,β-catenin abnormal expression and MMP-7 overexpression may be considered as two useful markers for determining metastasis and prognosis of human pancreatic cancer.     

Effects of vitamin E succinate on the expression of Fas and PCNA proteins in human gastric carcinoma cells and its clinical significance

AIM:To investigate the effects of vitamin E succinate (VES)on the expression of Fas and PCNA proteins as well as itsclinical significance in human gastric carcinoma,and toexplore the mechanism of VES-induced inhibition of gastriccarcinoma cell growth.METHODS:Immunohistochemical methods were used todetect Fas and PCNA expression both in human gastric cancerSGC-7901 cells treated with VES at different doses and inhuman gastric carcinoma tissues.RESULTS:After the SGC-7901 cells were treated with VESat 5,10,20 mg/L for 48 h,the positive rates of Fas expressionwere 16%,27% and 48%,respectively,significantly increasedcompared to that of control group (P<0.05);while the positiverates of PCNA expression in groups treated with differentdoses of VES were 20%,18% and 7%,respectively,whichwere significantly decreased compared to that of the controlgroup (P<0.05).In human gastric carcinoma tissues,the Faspositive expression rate was 42.4%(25/59),which declinedwith the decrease in the degree of tumor differentiation(P<0.05) and with the existence of lymph node metastasis(P<0.001).While the PCNA positive expression rate was91.5%(54/59),no relationship was observed between PCNAexpression and clinicopathologic parameters.CONCLUSION:VES inhibited the growth of gastric cancercells by inducing Fas expression and inhibiting PCNA expression.It is,therefore,considered that the expression of Fas andPCNA genes,through tumor cell apoptosis and proliferation,respectively,may be useful as a clinical predictive index inthe application of VES to gastric carcinoma therapy,whereas Fas may be of more value than PCNA.     

Up-regulation of PIK3CA promotes metastasis in gastric carcinoma

AIM:To explore expressions of PIK3CA in the progression of gastric cancer from primary to metastasis and its effects on activation of phosphatidylinositol 3-kinase(PI3K)/Akt pathway.METHODS:mRNA and protein levels of PIK3CA were assessed,respectively,by real-time quantitative polymerase chain reaction and immunohistochemistry in specimens of normal gastric mucosa,primary foci and lymph node and distant metastasis of gastric cancer.Akt and phosphorylated Akt protein were also examined by Western blotting in these tissues,in order to analyze the effect of PIK3CA expression level changes on the activation of PI3K/Akt signaling pathway.RESULTS:PIK3CA mRNA in lymph node metastasis were approximately 5 and 2 folds higher,respectively,than that in the corresponding normal gastric mucosa and primary gastric cancer tissues(P<0.05),while no statistical significance was found compared with distant metastasis.Immunohistochemically,PIK3CA protein expression was discovered in 7(35%)specimens of 20 primary foci vs 10(67%)of 15 of lymph node metastasis or 11(61%)of 18 of distant metastasis(35%vs 67%,P=0.015;35%vs 61%,P=0.044).With the increased level of PIK3CA expression,the total Akt protein expression remained almost unchanged,but p-Akt protein was upregulated markedly.CONCLUSION:Increased expression of PIK3CA is expected to be a promising indicator of metastasis in gastric cancer.Up-regulation of PIK3CA may promote the metastasis of gastric cancer through aberrant activation of PI3K/Akt signaling.     

Ubiquitin-like modifier activating enzyme 2 promotes cell migration and invasion through Wnt/β-catenin signaling in gastric cancer

AIM To investigate the function and mechanism of ubiquitinlike modifier activating enzyme 2(Uba2) in progression of gastric cancer(GC) cells.METHODS Uba2 level in patients with GC was analyzed by Western blotting and immunohistochemistry. MTT and colony formation assays were performed to examine cell proliferation.Flow cytometry was used for cell cycle analysis.Wound healing and Transwell assays were conducted to examine the effects of Uba2 on migration and invasion.Expression levels of cell cycle-related proteins, epithelial-mesenchymal transition(EMT) biomarkers, and involvement of the Wnt/β-catenin pathway was assessed by Western blotting. Activation of the Wnt/β-catenin pathway was confirmed by luciferase assay.RESULTS Uba2 expression was higher in GC than in normal tissues.Increased Uba2 expression was correlated with tissue differentiation, Lauren's classification, vascular invasion,and TNM stage, as determined by the analysis of 100 GC cases(P 0.05). Knock-down of Uba2 inhibited GC cell proliferation, induced cell cycle arrest, and altered expression of cyclin D1, P21, P27, and Bcl-2, while upregulation of Uba2 showed the opposite effects. The wound healing and Transwell assays showed that Uba2 promoted GC cell migration and invasion. Western blotting revealed alterations in EMT biomarkers, suggesting the role of Uba2 in EMT. Furthermore, the luciferase reporter assay indicated the involvement of the Wnt/β-catenin signaling pathway as a possible modulator of Uba2 oncogenic functions.CONCLUSION Uba2 plays a vital role in GC cell migration and invasion,possibly by regulating the Wnt/β-catenin signaling pathway and EMT.     

Blocking effects of siRNA on VEGF expression in human colorectal cancer cells

AIM:To investigate the expression of vascular endothelial cell growth factor (VEGF) and its receptors Fmslike tyrosine kinase 1 (FLT-1) and fetal liver kinase 1 (FLK-1) in colorectal carcinoma (CRC),and the blocking effects of small interfering RNAs (siRNAs) on VEGF expression in human colorectal cancer HCT116 cells.METHODS:Immunohistochemical staining for VEGF,FLT-1 and FLK-1 proteins was performed in 82 cases of CRC and 14 normal colorectal mucosae.A siRNA targeting VEGF was synthesized and transfected into HCT116 cells using lipofectamine 2000.Immunocytochemical staining and Western blotting analyses were performed to detect the expression of VEGF protein.The suppressive effect of the siRNA on cell proliferation was detected using the 3-(4,5-dimethylthiazol-2-yl)2,5-diphenyltertrazolium bromide (MTT) assay.Cellular apoptosis was detected using flow cytometry (FCM).RESULTS:The expression of VEGF,FLT-1 and FLK-1 in tumor tissues was significantly higher than that in normal tissues (P=0.008,P=0.000,P=0.000).The expression of VEGF was positively correlated with both lymph node metastasis and clinical stage (P=0.009 and P=0.025,respectively).Immunocytochemistry showed that the expression of VEGF was weakly positive and Western blotting indicated a significant reduction in VEGF-siRNA cell protein levels.VEGF-siRNA cell growth inhibition was assessed by the MTT assay,and the tumor cell proliferation rate was significantly different at 24,48,and 72 h after transfection.FCM results showed that the VEGF-siRNA group had an apparent aneuploid peak.CONCLUSION:VEGF,FLT-1 and FLK-1 are associated with colorectal carcinogenesis.siRNA silencing of the VEGF gene suppresses proliferation,and induces apoptosis in HCT116 cells.The results suggest that VEGF may be a new gene therapy target for colorectal cancer.     

β-catenin up-regulates the expression of cyclinD1, c-myc and MMP-7 in human pancreatic cancer： Relationships with carcinogenesis and metastasis

AIM: To investigate whether abnormal expression of β-catenin in conjunction with overexpression of cyclin D1, c-myc and matrix metalloproteinase-7 (MMP-7) correlated with the carcinogenesis, metastasis and prognosis of pancreatic cancer, and to analyze the relationship of β-catenin expression with cyclinD1, c-myc and MMP-7 expression.METHODS: Using immunohistochemistry, we examined the expression of β-catenin, cyclinD1, c-myc and MMP-7 in 47 pancreatic adenocarcinoma tissues, 12 pancreatic intraepithelial neoplasia (PanIN) and 10 normal pancreases,respectively. Proliferation cell nuclear antigen was also tested as the index of proliferative activity of pancreatic cancer cells.RESULTS: In 10 cases of normal pancreatic tissues,epithelial cells showed equally strong membranous expression of βcatenin protein at the cell-cell boundaries,but the expression of cyclin D1, c-myc and MMP-7 was negative. The expression of βcatenin, cyclinD1, c-myc and MMP-7 in PanIN and pancreatic adenocarcinoma tissues had no significant difference [6/12 and 32/47 (68.1%),6/12 and 35/47 (74.5%), 5/12 and 33/47 (70.2%), 7/12 and 30/47 (63.8%), respectively]. The abnormal expression of β-catenin was significantly correlated to metastasis and one-year survival rate of pancreatic cancer, but had no relation with size, differentiation and cell proliferation.The expression of cyclinD1 was correlated with cell proliferation and extent of differentiation, but not with size,metastasis and one-year survival rate of the pancreatic ancer. The expression of c-myc was not correlated with ize, extent of differentiation, metastasis and 1-year urvival rate, but closely with cell proliferation of pancreatic ancer. The overexpression of MMP-7 was significantly ssociated with metastasis and 1-year survival rate of ancreatic cancer, but not with size, extent of differentiation and cell proliferation. There was a highly significant positive association between abnormal expression of β-catenin and overexpression of cyclinD1, c-myc and MMP-7 not only in PanIN (r = 1.000, 0.845, 0.845), but also in pancreatic cancer (r = 0.437, 0.452, 0.435).CONCLUSION: The abnormal expression of β-catenin plays a key role in the carcinogenesis and progression of human pancreatic carcinoma by up-regulating the expression of cyclinD1, c-myc and MMP-7, resulting in the degradation of extracellular matrix and uncontrolled cell proliferation and differentiation, β-catenin abnormal expression and MMP-7 overexpression may be considered as two useful markers for determining metastasis and prognosis of human pancreatic cancer.     

Effect of BRM S1 expression on proliferation,migration and adhesion of mouse forestomach carcinoma

Objective: To discuss the ef ect of BRMS1 on the proliferation, migration and adhesion of mouse forestomach carcinoma(MFC). Methods: The constructed p CMV-myc-BRMS1 recombinant plasmid and blank plasmid were transfected into mouse forestomach carcinoma. MTT method was employed to measure the activity of gastric cancer cell; the scratch assay and Transwell assay to measure the migration and invasion of gastric cancer cell; the adhesion assay to measure the adhesion of gastric cancer cell; while the Western blot assay to measure the expression of The NF-毷B signal pathway, downstream matrix metalloproteinase(MMP-2), MMP-9 and osteopontin and E-cadherin in the gastric cancer cell. Besides, the transplanted animal model of gastric cancer in mice was constructed to measure the size of tumor xenograft. Results: Results of MTT assay showed that, compared with the empty vector control group, the activity of gastric cancer cell was not af ected in the BRMS1 transfection group. The improved expression of BRMS1 could inhibit the adhesion, migration and invasion of gastric cancer cell(P0.01). Besides, compared with the empty vector control group, the phosphorylation of NF-毷B p65 and I毷Bα was reduced in the BRMS1 transfection group, with the decreased expression of MMP 2, MMP 9 and osteopontin and the increased expression of E-cadherin(P0.01). Results of animal experiment also showed that the expression of BRMS1 did not af ect the transplanted tumor. Conclusions: The expression of BRMS1 can signii cantly inhibit the adhesion, migration, invasion and metastasis of MCF gastric cancer cell, which is related to The NF-毷B signal pathway.     

Relationship between preoperative staging by endoscopic ultrasonography and MMP-9 expression in gastric carcinoma

AIM： To investigate the relationship between the staging by endoscopic ultrasonography （EUS） and the expression of carcinoma metastasis associated gene in the patients with gastric carcinoma.
METHODS： Sixty-three patients with gastric cancer were diagnosed by electric gastroscopy and EUS. The preoperative staging of gastric cancer was measured by EUS and compared with pathologic staging and MMP-9 expression. Peripheral serum level of MMP-9 was measured with enzyme-linked immunosorbent assay （ELISA）, while the expression of MMP-9 protein was tested with immunohistochemistry and hybridization in situ in the gastric carcinoma tissues.
RESULTS： The total accuracy of EUS in estimating invasive depth of gastric cancer was 80.95%, while that in estimating lymphatic metastasis was 73.02%.Serum MMP-9 levels were consistent with the expression of MMP-9 protein and MMP-9 mRNA in tissue, a result closely correlated with invasive degree, staging with EUS and lymphatic metastasis in gastric cancer （P 〈 0.05）.The total accuracy of estimating invasive depth in gastric cancer was 95.22% using both EUS and MMP-9.
CONCLUSION： The MMP-9 level of preoperative serum presents the reference value for preoperative staging by EUS in the patients with gastric cancer. When serum MMP-9 level in gastric cancer is significantly high,physicians should pay closer attention to the metastasis which reaches the serosa or beyond. Combining EUS and MMP-9 improves the accuracy in deciding the invasion and metastasis in the patients with gastric carcinoma.