MiR-125b promotes cell migration and invasion by targeting PPP1CA-Rb signal pathways in gastric cancer, resulting in a poor prognosis.

BACKGROUND: MiR-125b functions as an oncogene in many cancers; however, its clinical significance and molecular mechanism in gastric cancers have never been sufficiently investigated. Here, we elucidated the functions and molecular regulated pathways of MiR-125b in gastric cancer. METHODS: We investigated MiR-125b expression in fresh tissues from 50 gastric cancer patients and 6 gastric cancer cell lines using RT-PCR, and explored its prognostic value by hybridizing MiR-125b in situ for 300 clinical gastric tumor tissues with pathological diagnosis and clinical parameters. The effects of MiR-125b on gastric cancer cells and downstream target genes and proteins were analyzed by MTT, transwell assay, RT-PCR, and western blot on the basis of silencing MiR-125b in vitro. Luciferase reporter plasmid was constructed to demonstrate MiR-125b's direct target. RESULTS: MiR-125b was upregulated in gastric cancer tissues and cell lines, and significantly promoted cellular proliferation, migration, and invasion by downregulating the expression of PPP1CA and upregulating Rb phosphorylation. MiR-125b expression was significantly correlated with tumor size and depth of invasion, lymph nodes, distant metastasis, and TNM stage. The high-MiR-125b-expression group had a significantly poorer prognosis than the low-expression group (P < 0.05) in stages I, II, and III, and the 5-year survival rate in of the high-expression group was significantly lower than that of the low-expression group. CONCLUSIONS: MiR-125b functions as an oncogene by targeting downregulated PPP1CA and upregulated Rb phosphorylation in gastric cancer. MiR-125b not only promotes cellular proliferation, migration, and invasion in vitro, but also acts as an independent prognostic factor in gastric cancer.

MicroRNA-10b promotes migration and invasion through Hoxd10 in human gastric cancer.

BACKGROUND: This study aims to investigate the effect of miR-10b overexpression on cancer cell proliferation, migration, invasion, and Hoxd10 expression. METHODS: The effect of miR-10b on proliferation, migration, and invasion of MKN-28, BGC-823, and SGC-7901 cells and the expression of Hoxd10 protein in SGC-7901 and BGC-823 cells were detected following transfection of miR-10b inhibitor or Negative Control B. Expression of Hoxd10 protein in 436 paraffin-embedded cancer tissues was also investigated. RESULTS: miR-10b was significantly upregulated in AGS, MKN-28, BGC-823, HCG-27, SGC-7901, and MKN-45 cell lines, miR-10b inhibitor significantly inhibited proliferation and migration of MKN-45, BGC-823 and SGC-7901 cells 48 h after transfection, while Hoxd10 protein in these cells lines had increased 72 h after transfection. Hoxd10 was highly expressed in gastric cancer and correlated with size of tumor, Lauren classification, depth of invasion, lymph node and distant metastasis, Tumor-Node-Metastasis (TNM) stage, and prognosis. CONCLUSIONS: miR-10b promotes migration and invasion through Hoxd10 in human gastric cancer cell lines and may play an important role in tumorigenesis, progression, and prognosis.

Abnormal expression of miR-301a in gastric cancer associated with progression and poor prognosis.

BACKGROUND AND OBJECTIVES: miR-301a is significantly overexpressed in many cancers. However, its expression and biological role in gastric cancer remain poorly understood. We investigated microRNA-301a (miR-301a) expression in gastric cancer and determined its effects on cancer cell behavior and its clinical significance in the development and progression of gastric cancer. METHODS: We determined miR-301a expression in gastric tumors and gastric cancer cell lines by reverse transcription-polymerase chain reaction. The effects of miR-301a on cell clone formation, migration, and invasion of HGC-27 and SGC-7901 cells were detected following transfection of an miR-301a inhibitor. miR-301a expression in a 304-tissue gastric cancer microarray was determined by in situ hybridization and its role in progression and prognosis was analyzed. RESULTS: miR-301a was upregulated in gastric tumor tissues and cell lines. Down-regulation of miR-301a significantly inhibited cell clone formation, migration, and invasion of HGC-27and SGC-7901 cells. Overexpression of miR-301a in primary gastric cancer tissues was associated with tumor size, invasion depth, lymph node metastasis, and TNM stage. CONCLUSIONS: miR-301a overexpression correlated with TNM stage and prognosis, suggesting that miR-301a is involved in cellular clone formation, migration, and invasion in vitro and may play an important role in the clinical progression and prognosis of gastric cancer.

Clinical utility of measuring expression levels of KAP1, TIMP1 and STC2 in peripheral blood of patients with gastric cancer.

BACKGROUND: We examined preoperative kinesin II-associated protein (KAP1), TIMP metallopeptidase inhibitor 1 (TIMP1) and stanniocalcin 2 (STC2) expression levels in patients with gastric cancers to assess their clinical application for diagnosing and monitoring diseases. METHODS: Real-time reverse transcription-polymerase chain reaction was used to detect the expression levels of KAP1, TIMP1, STC2, talin 2 (TLN2), sushi-repeat-containing protein, X-linked 2 (SRPX2) and secreted protein, acidic, cysteine-rich (SPARC) in the patients' peripheral blood karyocytes. The data were analyzed with receiver operating characteristics (ROC) curves. RESULTS: A total of 112 patients with gastric cancer, 42 patients with recurrence and 107 healthy volunteers were recruited. There were significant correlations between KAP1, TIMP1 and STC2 levels, and TNM tumor stages and distant metastases. The area under the ROC curves (AUC) of KAP1 was 0.803 ± 0.040 (P = 0.0001), the AUC of TIMP1 was 0.767 ± 0.043 (P = 0.0001) and the AUC of STC2 was 0.769 ± 0.045 (P = 0.0001), thus differentiating preoperative gastric cancer patients from healthy volunteers by ROC curve analysis. The AUC of STC2 was 0.739 ± 0.070 (P = 0.004) and the AUC of KAP1 was 0.418 ± 0.088 (P = 0.319), thus differentiating recurrence of gastric cancer from healthy volunteers by ROC curve analysis. High TIMP1 and STC2 expression levels were suspected to be poor prognostic factors of disease recurrence in patients with gastric cancer. CONCLUSIONS: KAP1, TIMP1 and STC2 expression levels may be potential biomarkers for the screening, diagnosis, prognosis and surveillance of gastric cancer.

[Detection and clinicopathologic significance of microsatellite alteration and p53 mutation of neuroendocrine cells in colorectal adenocarcinoma].

OBJECTIVE: To study the possible clonal origin of neuroendocrine cells in colorectal adenocarcinoma. METHODS: Twenty-six microsatellite loci were screened using laser capture microdissection, DNA extraction and whole genome amplification. Microsatellite instability (MSI) and loss of heterozygosity (LOH) in adenocarcinoma cells and neuroendocrine cells amongst 30 cases of colorectal carcinoma with neuroendocrine differentiation were detected using polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP)-silver staining. The mutation status of p53 was evaluated by PCR-sequencing. The clonal origin of neuroendocrine cells in colorectal adenocarcinoma was determined. RESULTS: Amongst the 30 cases studied, the prevalence of MSI was 16.9% while that of LOH was 8.5%. The rate showed no statistically significant difference between adenocarcinoma cells and neuroendocrine cells. In 6 cases, the microsatellite alteration was entirely consistent. In 23 cases, the rate of microsatellite alteration consistency was greater than that of inconsistency. In 1 case, the consistency and inconsistency rates were identical. There was statistically significant difference between consistency and inconsistency of microsatellite alteration. The prevalence of p53 mutation was 16.7% which was the same for both adenocarcinoma cells and neuroendocrine cells. CONCLUSIONS: Adenocarcinoma cells and neuroendocrine cells in colorectal adenocarcinoma with neuroendocrine differentiation have similar biologic changes. It is likely that they are of identical origin.

Upregulated expression of ADAM17 is a prognostic marker for patients with gastric cancer.

OBJECTIVE: This study was designed to evaluate the expression and prognostic significance of A Disintegrin and Metalloproteinase17 (ADAM17) protein in patients with gastric cancer. BACKGROUND: Tumor invasion and metastasis are primary causes for treatment failure or death among cancer patients. ADAM17 is a multidomain transmembrane glycoprotein involved in the release of several ligands that were shown to promote tumor formation and progression. Elevated expression of ADAM17 was detected in a number of human cancers and was associated with poor progression and prognosis of the diseases. In gastric cancer, however, the expression and prognostic significance of ADAM17 has not been fully elucidated. METHODS: The expressions of ADAM17 and extracellular matrix metalloproteinase inducer (EMMPRIN), a protein implicated in tumor invasion and metastasis, were detected using the tissue microarray technique and immunohistochemical EnVision method and compared with clinicopathological parameters of patients with gastric cancer. RESULTS: The expressions of ADAM17 and EMMPRIN were upregulated in gastric cancer lesions compared with their expressions in adjacent non-cancerous tissues (P < 0.01). High expression of ADAM17 was detected in 35.78% (156/436) of patients with gastric cancer and positively correlated with the expression of EMMPRIN (r = 0.738, P < 0.01). ADAM17 expression was associated with a number of clinicopathological parameters including depth of invasion and TNM stage of the tumor (P < 0.05). In each TNM stage, patients with high ADAM17 expression had a longer mean survival time than those with low expression (P < 0.05). Particularly, the mean survival time of stage II gastric cancer patients with low ADAM17 expression was longer than that of stage I patients with high ADAM17 expression (P < 0.01). Multivariate survival analysis suggested that, along with other parameters, ADAM17 and EMMPRIN expression were independent prognostic factors for patients with gastric cancer. CONCLUSIONS: ADAM17 was implicated in the progression of gastric cancer. On the basis of the TNM stage, detection of ADAM17 expression will be helpful for predicting prognosis of gastric cancer.

Expression and prognostic significance of CEACAM6, ITGB1, and CYR61 in peripheral blood of patients with gastric cancer.

BACKGROUND: We examined CEACAM6, ITGB1, and cyr61 concentrations from patients with gastric cancers (GCs) to assess their clinical application for diagnosing and monitoring diseases. METHODS: Real-time reverse transcription-polymerase chain reaction was used to detect the expressions of CEA, CEACAM6, ITGB1, IGF1R, CK20, cyr61, and S100A4 in peripheral blood karyocyte from 82 patients with GC, 24 patients with recurrence, and 37 healthy volunteers. Receiver operating characteristics (ROC) curves were constructed. RESULTS: There were significant association between these CEACAM6, ITGB1, and cyr61 and TNM Stages and distant metastasis. The AUC of CEACAM6 was 0.884 ± 0.044 (P = 0.0001), the AUC of cyr61 was 0.833 ± 0.047 (P = 0.0001), and the AUC of ITGB1 was 0.838 ± 0.042 (P = 0.0001) by differentiating preoperative GC patients from healthy volunteers from ROC curve analysis. The AUC of CEACAM6 was 0.761 ± 0.066 (P = 0.001), the AUC of CYR61 was 0.762 ± 0.063 (P = 0.001), and the AUC of ITGB1 was 0.824 ± 0.051 (P = 0.0001), by differentiating recurrence of GC from healthy volunteers from ROC curve analysis. CONCLUSION: The method of detecting the expression of CEACAM6, ITGB1, and CYR61 in peripheral blood of GC patients was more sensitive than CEA, IGF1R, CK20, and S100A4 for the early diagnosis of metastasis and recurrence.

ADAM 10 is associated with gastric cancer progression and prognosis of patients.

BACKGROUND: The metalloproteinase domain-containing protein 10 (ADAM 10) has been implicated in the development and progression of gastric cancer. METHODS: Expression of ADAM 10 and C-erbB-2 were examined immunochemically in 436 clinicopathologically characterized gastric cancer cases. RESULTS: Protein levels of ADAM 10 and C-erbB-2 were up-regulated in gastric cancer lesions compared with adjacent non-cancerous tissues. Positive expression of ADAM 10 correlated with age, size of tumor, location of tumor, depth of invasion, vessel invasion, lymph node, and distant metastasis and TNM stage, and also with expression of C-erbB-2. In stages I, II, and III, the 5-year survival rate of patients with high ADAM 10 expression was significantly lower than in patients with low expression. However, in stage IV, ADAM 10 expression did not correlate with the 5-year survival rate. Further multivariate analysis suggests that up-regulation of ADAM 10 and C-erbB-2 were independent prognostic indicators for the disease, along with depth of invasion, lymph node and distant metastasis and TNM stage. CONCLUSION: Expression of ADAM 10 in gastric cancer is significantly associated with lymph node and distant metastasis, high C-erbB-2 expression, and poor prognosis. ADAM 10 and C-erbB-2 proteins could be useful markers to predict tumor progression and prognosis.

High-level expression of S100A4 correlates with lymph node metastasis and poor prognosis in patients with gastric cancer.

BACKGROUND: The present study investigated the clinical significance of S100 calcium binding protein A4 in the development, progression, and metastasis of gastric cancer. METHODS: Tumor tissue, adjacent normal tissue, and lymph node and peritoneal metastases were obtained from patients with gastric cancer, and their gene expression profiles were analyzed by Affymetrix GeneChip HG-U133A2.0 array. The expression of S100A4 was detected by real-time quantitative reverse-transcription polymerase chain reaction (RT-PCR) in gastric tumor tissue and lymph node and peritoneum metastasis. Immunohistochemistry was employed to analyze S100A4 expression in 436 clinicopathologically characterized gastric cancer cases and in corresponding distant metastases from 61 patients. RESULTS: A total of 434 genes and 169 expressed sequence tags were upregulated by at least twofold in the tumor tissue. The expression of S100A4 in lymph node and peritoneal metastases was significantly higher than that in gastric tumor tissue. The expression of S100A4 messenger RNA (mRNA) or protein differed significantly among gastric tumor tissue, matched normal gastric mucosa, and lymph node and peritoneal metastases. Further multivariate analysis suggested that depth of invasion, lymph node and distant metastases, tumor-node-metastasis (TNM) stage, and upregulation of S100A4 were independent prognostic indicators for the disease. CONCLUSION: Gene expression profiles are a useful way to perform simultaneously large-scale analysis of the expression level of thousands of genes. Expression of S100A4 in gastric cancer is associated significantly with lymph node and distant metastases, and poor prognosis. S100A4 may be a useful marker to predict development, progression, and metastasis of gastric cancer.

L1, ß-catenin, and E-cadherin expression in patients with colorectal cancer: correlation with clinicopathologic features and its prognostic significance.

OBJECTIVE: Currently, there are no clinically used/routine biomarkers that accurately predict whether colorectal cancer (CRC) patients will or will not respond to adjuvant chemotherapy. The aim of this study was to investigate L1, ß-catenin, and E-cadherin expression in patients with CRC and their relationship to tumor progression, and to identify patients who will respond to chemotherapy. METHODS: A total of 142 patients who underwent surgical treatment for CRC were chosen retrospectively. The samples from these patients were analyzed by immunohistochemistry. SPSS-14 program package was used for statistical calculation. RESULTS: Expression of L1, ß-catenin, and E-cadherin were found to be strongly associated with invasion and metastasis of CRC. Cox multivariate analysis results indicated that L1 expression and stage of Dukes could be considered as the independent prognostic factors for survival. Furthermore, our study found that the 5-year survival rate was the significantly associated with the expression of L1, ß-catenin (normal and ectopic expression), and E-cadherin for Dukes' stage B (P < 0.01) patients. However, no such result was found for Dukes' stage A (P > 0.05) and C (P > 0.05) patients. CONCLUSION: Our study provided reference for identifying patients who need adjuvant chemotherapy. L1, ß-catenin, and E-cadherin could be considered as biomarkers to predict whether CRC patients will or will not respond to adjuvant chemotherapy.

[Relationship between E-CD and Snail expressions and tumor invasion, metastasis and prognosis in colorectal cancer].

OBJECTIVE: To study the E-CD and Snail expressions in colorectal cancer and their relationship with colorectal cancer invasion, metastasis and prognosis. METHODS: Immunohistochemical staining (EnVision) was used to detect the E-CD and Snail expressions in 30 normal colorectal mucosa, 30 colorectal adenoma and 142 colorectal cancer tissues. RESULTS: E-CD in the normal colorectal mucosa was strongly positive expressed (90.0%), significantly higher than that in colorectal adenomas (63.3%) and colorectal cancer tissues (41.5%). E-CD expression was significantly related to tumor differentiation, invasion depth, vascular invasion, lymph node metastasis and Dukes' stage (P < 0.05), but not to the patients' age, gender, tumor size and tumor histological type (P > 0.05). The 1-, 3- and 5-year survival rates of the E-CD positive patients with colorectal cancer were significantly higher than that in E-CD negative patients. The positive expression rate of Snail in colorectal cancer tissues (52.1%) was significantly higher than that in normal colorectal mucosa (6.7%) and colorectal adenomas (26.7%, P < 0.05). The snail expression was significantly correlated to tumor histological type, differentiation, invasion depth, vascular invasion, lymph node metastasis and Duke's stage (P < 0.05), but not to patients' age, sex and tumor size (P > 0.05). The 1-, 3- and 5-year survival rates of Snail negative patients with colorectal cancer was significantly higher than that in patients with positive expression (P < 0.05). The expressions of E-CD and Snail in colorectal cancer tissues were inversely correlated (P < 0.05). Cox multivariate analysis showed that E-CD and Snail can be used as independent prognostic indicators (P < 0.05). CONCLUSION: E-CD and Snail expressions in colorectal cancer are related to the tumor invasion, metastasis and prognosis. Low expression of E-CD and high expression of Snail are related to the advanced stage, and poor prognosis in colorectal cancer patients. E-CD and Snail can be used as independent prognostic indicators.

[Expression of matrix metalloproteinase-2 and insulin-like growth factor-1 in gastric carcinoma and their clinicopathological significance].

OBJECTIVE: To investigate the expression of matrix metalloproteinase-2 (MMP-2) and insulin-like growth factor-1 (IGF-1) in gastric carcinoma and their clinicopathological significance. METHODS: Expressions of MMP-2 and IGF-1 were examined by using immunohistochemical SP staining and cross-compared with clinicopathological features of gastric carcinoma. RESULTS: High expression of MMP-2 and IGF-1 were observed in 70.4% (307/436) and 49.5% (216/436) of gastric carcinoma tissues respectively, significantly higher than those in non-tumor gastric mucosa (3.3% and 5.4%, respectively; all P < 0.05). The high expression rate of MMP-2 and IGF-1 were significantly associated with the patient age, tumor size, tumor location, Lauren classification, TNM staging, depth of tumor infiltration, presence of vessel invasion, lymph node and distant metastasis (all P < 0.05). In addition, the expression of MMP-2 was positively linked with the expression level of IGF-1 (P < 0.05). Univariate analysis showed that high expression of MMP-2, was significantly associated with poor prognosis of tumor of TNM stage I and II (all P < 0.05), high expression of IGF-1 was significantly correlated with poor prognosis of patients with TNM stage I, II and III tumor (all P < 0.05). Cox multivariate analysis indicated that the high expressions of MMP-2 and IGF-1 could be independent prognostic indices for gastric carcinoma. CONCLUSIONS: High expression of MMP-2 and IGF-1 proteins are significantly correlated with the invasion and metastasis of gastric carcinoma, it is helpful to simultaneously detect the expressions of MMP-2 and IGF-1 proteins in predicting prognosis of patients with gastric carcinoma.

Screening and identification of novel B cell epitopes in human heparanase and their anti-invasion property for hepatocellular carcinoma.

BACKGROUND: The aim of this study was to screen and identify novel B cell epitopes within the human heparanase protein and to investigate the impact of self-developed anti-heparanase polypeptide antibodies on growth and invasion of HCCLM6 human hepatocellular carcinoma cells in vitro. METHODS: The flexible regions of secondary structure and the B cell epitopes of the human heparanase amino acid sequence were predicted by DNAStar and Bcepred software.The multiple antigenic peptides (MAP) of the epitopes were synthesized in eight-branched form. Rabbits were immunized with the eight-branched MAPs mixed with the universal T-helper epitope human IL-1beta peptide (VQGEESNDK, amino acid 163-171). The immunogenicity of the synthesized peptides was evaluated by ELISA, western blot and immunohistochemistry. The impact of the self-developed rabbit anti-heparanase polyclonal antibodies on growth and invasion ability of HCCMLM6 cells were analyzed in a cell culture model. The cells were first treated with one of the three antibodies, respectively, and then measured by using MTT, flow cytometry, plate clone formation, invasion assay and heparan sulfate degrading enzyme assay. RESULTS: The three amino acid sequences 1-15 (MAP1), 279-293 (MAP2), and 175-189 (MAP3) in the large subunit of the human heparanase protein were predicted as its most potential epitopes. ELISA, western blot and immunohistochemistry analysis showed that all three MAPs were capable to induce high titer of serum antibodies. Antibodies induced by MAP1 and MAP2 were high specific. Furthermore, anti-MAP2 antibodies showed the strongest avidity towards liver cancer tissues. Under the treatment with the three anti-heparanase antibodies, respectively, the growth, cell cycle and clone formation of the cells remained unchanged when compared with a treatment with normal rabbit IgG. However, an inhibition of cell invasiveness and heparanase activity could be detected under the treatment with anti-MAP1- or anti-MAP2-antibody (with a terminal concentration of 100 mug/ml). The cell invasiveness was decreased by 54 and 38%, respectively, the heparanase activity by 43 and 39%, respectively. CONCLUSION: The multiple antigenic peptides MAP1 (AC 1-15) and MAP2 (AC 279-293) may be the dominant B cell epitopes in the human heparanase protein. The induced polypeptide antibodies can effectively inhibit the heparanase activity of HCCLM6 liver cancer cells and therefore influence their invasion ability, which provides a theoretic basis for the development of anti-heparanase antibodies and their clinical use as vaccine.

[Surgical treatment for primary gastrointestinal stromal tumors: a report of 73 cases].

OBJECTIVE: To investigate the effects and prognosis of surgical treatment in primary gastrointestinal stromal tumors (GIST). METHODS: The clinicopathological data of 73 patients with primary GIST underwent operation from April 1997 to December 2007 was retrospectively analyzed, and the prognosis was evaluated too. RESULTS: Among the 73 cases, 68 cases received complete tumor resection, among which 12 cases underwent laparoscopic operation; while palliative resection and biopsy only were carried out in the other 5 cases. There was significant difference in survival rate between the two groups (P = 0.000). The 1-, 3-, 5-year survival rates of the 66 cases had been followed up was 91.0%, 78.2% and 74.1%, respectively. The malignancy risk grades of GIST was related to the survival rates on statistical analysis (P = 0.002). Significant differences were found in the survival rates between the patients with very low grade, low grade and high grade malignancy tumors (P = 0.012, 0.002). CONCLUSIONS: Complete tumor resection should be emphasized in primary GIST, and more attention should be paid to the initial surgical treatment. Extended surgical resection is required for tumors of higher malignancy risk. The indications of laparoscopic surgery in GIST should be selected with caution for tumor complete resection.

Management of Combined Atlas Fracture with Type II Odontoid Fracture: A Review of 21 Cases.

PURPOSE: To evaluate the therapeutic effects of combined atlas fracture with type II (C1-type II) odontoid fractures and to outline a management strategy for it. PATIENTS AND METHODS: Twenty three patients with C1-type II odontoid fractures were treated according to our management strategy. Nonoperative external immobilization in the form of cervical collar and halo vest was used in 13 patients with stable atlantoaxial joint. Surgical treatment was early performed in 10 patients whose fractures with traumatic transverse atlantal ligament disruption or atlantoaxial instability. The visual analog scale (VAS), neck disability index (NDI) scale, and American Spinal Injury Association (ASIA) scale at each stage of followup were then collected and compared. RESULTS: Compared to pretreatment, the VAS score, NDI score, and ASIA scale were improved among both groups at followup evaluation after treatment. However, in the nonsurgical group, one patient (1/11) developed nonunion which required surgical treatment in later stage and one patient (1/13) with halo vest immobilization had happened pin site infection. Two patients of the surgical group (2/11) had appeared minor complications: occipital cervical pain in one case and cerebrospinal fluid leakage in one case. Two patients (2/23) were excluded from nonsurgical treatment group because their followup period was less than 12 months. Twenty one patients were followed up regularly with an average of 23.9 months (range 15-45 months). CONCLUSIONS: We outlined our concluding management principle for the treatment of C1-type II odontoid fractures based on the nature of C1 fracture and atlantoaxial stability. The treatment principle can obtain satisfactory results for the management of C1-type II odontoid fractures.

Correlation of integrin beta3 mRNA and vascular endothelial growth factor protein expression profiles with the clinicopathological features and prognosis of gastric carcinoma.

AIM: To investigate integrin beta3 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 beta3 mRNA and immunohistochemistry of VEGF and CD34 protein were performed on samples from 118 patients with gastric cancer. RESULTS: The positive rate of integrin beta3 mRNA in non-tumor gastric mucosa (20%) was significantly lower than that of the gastric cancer tissue (52.5%, c2 = 10.20, P < 0.01). In patients of infiltrating type, stage T(3)-T(4), vessel invasion, lymphatic metastasis, hepatic or peritoneal metastasis, the positive expression rates of integrin beta3 mRNA were significantly higher than those in patients of expanding type (P < 0.01), stage T(1)-T(2) (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 T(3)-T(4), 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 T(1)-T(2) (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 T(3)-T(4), 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 T(1)-T(2) (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 beta3 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 MVD (P < 0.05). The mean survival period in patients with positive expression of integrin beta3 mRNA and VEGF, and MVD > or = 54.9/mm(2) was significantly shorter than that in patients with negative expression of integrin beta3 mRNA (P < 0.05) and VEGF (P < 0.01), and MVD < 54.9/mm(2) (P < 0.01). Five-year survival rate in patients with positive expression of integrin beta3 mRNA and VEGF, and MVD > or = 54.9/mm(2) was significantly lower than those with negative expression of integrin beta3 mRNA (P < 0.05), VEGF (P < 0.05), and MVD < 54.9/mm(2) (P < 0.01). CONCLUSION: Integrin beta3 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.

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 beta3 were evaluated by immunohistochemical study in a total of 118 gastric carcinomas and 20 non-tumor 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 beta3 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 beta3 (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 beta3 (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 beta3 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 beta3 are significantly correlated with the invasion and metastasis of gastric carcinoma, and they are highly correlated with each other. Therefore they may serve as important prognostic markers of gastric carcinoma.

[mRNA expression of basic fibroblast growth factor and hepatocyte growth factor in gastric carcinoma and significance thereof].

OBJECTIVE: To investigate the mRNA expression of basic fibroblast growth factor (bFGF) and hepatocyte growth factor (HGF) in gastric carcinoma and correlation thereof with intratumoral microvascular density (IMD), proliferating cell nuclear antigen labeling index (PCNA LI), tumor invasion and metastasis, and patients' survival. METHODS: In situ hybridization and immunohistochemistry were used to test the expression of bFGF and HGF mRNA and protein expression of CD34 and PCNA in 118 specimens of gastric carcinoma. RESULTS: The positive rates of bFGF mRNA and HGF mRNA were 57.6% and 52.5% respectively; the positive expression rates of bFGF mRNA and HGF mRNA of the cases at the stage T3-T4, and those with vessel invasion, lymph node metastasis, and distant metastasis were all significantly higher than the positive expression rates of the cases at the stage T1-T2, and those without lymph node metastasis, vessel invasion, and distant metastasis (all P < 0.05). The mean IMD levels of the patients positive in bFGF and HGF mRNA expression were (46.63 +/- 13.96) and (46.73 +/- 13.34) pieces/0.72 mm2, both significantly higher than those of the patients negative in bFGF and HGF mRNA expression [(34.56 +/- 15.16) and (35.98 +/- 14.92) pieces/0.72 mm2) respectively, t = 4.62, P = 0.000, and t = 4.01, P = 0.000]. The PCNA LI of the of the patients positive in bFGF and HGF mRNA expression was (66.53 +/- 13.61) and (67.56 +/- 13.41) respectively, both significantly higher5 than those of patients negative in bFGF and HGF mRNA expression [(54.79 +/- 10.57) and (55.04 +/- 11.01) respectively, t = 5.83, P = 0.000, and t = 4.91, P = 0.000]. The 5-year survival rates of the patients negative in bFGF and HGF mRNA expression were 60.00% and 67.90% respectively, both significantly higher than those of the patients positive in bFGF and HGF mRNA expression (30.90% and 21.00% respectively, both P = 0.000). IMD, PCNA LI, and bFGF mRNA were all positively related to HGF mRNA (all P = 0.000). CONCLUSIONS: bFGF and HGF promote angiogenesis and proliferation of gastric cancer, and in involved in tumor invasion and metastasis. They can be used as markers of prognosis of gastric cancer in clinical practice.

[Relationship of cysteine-rich protein 61 and NF-kappaB expression with metastasis and prognosis of gastric carcinoma].

OBJECTIVE: To investigate the expression of cysteine-rich protein 61 (Cyr61) and NF-kappaB in gastric carcinoma and its correlation with the clinicopathologic features and survival time. METHODS: RT-PCR was used to validate and detect expression of Cyr61 mRNA in 53 gastric carcinoma specimens and 11 non-tumor gastric mucosa samples. Cyr61 and NF-kappaB protein levels expressed were detected using immunohistochemistry in 99 gastric carcinoma specimens and 25 non-tumor gastric mucosa samples. RESULTS: RT-PCR demonstrated that expression of Cyr61 mRNA was higher in the primary carcinoma (84.6%, 22/26) and the metastatic foci (88.9%, 24/27) than in the non-tumor control samples (5/11; P < 0.05, respectively). Cyr61 gene mRNA expression levels were (2.76 +/- 5.50) x 10(-5), (14.61 +/- 20.64) x 10(-5), and (18.46 +/- 26.38) x 10(-5) by 2(-DeltaCt) in the control mucosa samples, primary carcinomas and metastatic tissues respectively. The level was higher in the primary carcinomas and metastatic tissues than that of the non-tumor gastric mucosa (P < 0.05, respectively); however, there was no significant difference between the metastatic tissues and the primary carcinomas (P > 0.05). Immunohistochemistry revealed that of the 99 cases, there was a high expression of Cyr61 and NF-kappaB protein, 56.6% (56/99) and 55.6% (55/99) respectively. There was correlation of Cyr61 and NF-kappaB protein expressions with the depth of tumor and vascular invasion, as well as the development of lymph node metastasis, and TNM staging (P < 0.05, respectively), besides, the expression of NF-kappaB also correlated with the tumor diameter (P < 0.05). Cyr61 expression was positively correlated with NF-kappaB expression in gastric carcinoma (P < 0.05); the mean survival time in cases with a high expression level of Cyr61 and NF-kappaB protein was significantly shorter than those with a low expression level (P < 0.05). CONCLUSIONS: Expression of Cyr61 and NF-kappaB closely correlated with invasion and metastasis of gastric carcinoma. They may be considered as the biologic behavior indicators for gastric carcinoma.

Metastasis-associated protein 1 (MTA1) in gastric cancer tissues is positively associated with poorer prognosis.

BACKGROUND: The present study examined the clinical significance of metastasis-associated protein 1 (MTA1) in the progression and patient survival of gastric cancer. METHODS: Paraffin-embedded resected tissues of gastric cancer mucosa (n = 436) and adjacent normal mucosa (n = 92) were assessed immunohistochemically for MTA1 protein, and scored according to the percentage of cells positively stained for MTA1 combined with stain intensity. Associations between MTA1 staining scores and clinicopathological factors, including survival time, were evaluated. RESULTS: The staining scores for MTA1 were significantly higher in gastric cancer tissues than in matched normal tissues. MTA1 scores positively correlated with tumor size, depth of invasion, presence of lymph node metastasis, lymphatic involvement, venous invasion, distal metastasis, and advanced clinical staging. Patients with high MTA1 scores in gastric cancer tissues had a significantly lower five-year survival rate compared with patients with low MTA1 scores. The multivariate analysis indicated that MTA1 protein levels in resected gastric cancer tissues, as reflected by immunohistochemical staining, are an independent prognostic index of gastric carcinoma (P < 0.01). CONCLUSION: MTA1 immunopositivity was significantly associated with progression of gastric cancer, and may be helpful in gastric cancer prognosis.