Expression In Gastric Carcinoma Research Articles

Aberrant expression of EphA3 in gastric carcinoma: correlation with tumor angiogenesis and survival

EphA3, a member of the Eph receptor tyrosine kinases, plays important roles in tumor angiogenesis and progression. However, the function of EphA3 in solid tumors has not been widely studied. We aimed to explore EphA3 expression in gastric carcinoma and analyze its role as a potential prognostic factor. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to assess EphA3 mRNA in a normal gastric mucosa cell line and carcinoma cell lines. Immunohistochemistry for EphA3 and vascular endothelial growth factor (VEGF) was performed in 318 cases of gastric carcinoma. CD34 immunohistochemical staining was used for microvessel density (MVD) counting. Western blotting was used to analyze EphA3 expression in the cell lines and to determine the expression of EphA3 and VEGF in 75 cases of gastric carcinoma and matched normal mucosa. EphA3 mRNA and protein expression was significantly higher in gastric cancer than that in normal mucosa (all P < 0.001). EphA3 was significantly correlated with TNM stage and poor prognosis (all P < 0.001). Multivariate analysis showed that EphA3 had an independent effect on survival (P = 0.037). EphA3 was positively correlated with VEGF (P < 0.001), and MVD (P < 0.001). According to Western blot analysis, both EphA3 and VEGF expression were significantly higher in carcinoma than that in normal mucosa (all P < 0.001). A positive correlation was observed between EphA3 and VEGF expression in cancer (P < 0.001, r = 0.513). EphA3 may play important roles in the angiogenesis and prognosis of gastric carcinoma, and thus may become a useful target for therapeutic intervention and a potential indicator for clinical assessment of tumor prognosis.

Caveolin-1, E-cadherin and β-catenin in gastric carcinoma, precancerous tissues and chronic non-atrophic gastritis

To investigate the expressions of caveolin-1, E-cadherin and β-catenin in gastric carcinoma, precancerous gastric and chronic non-atrophic gastritis tissues, and evaluate the correlation of these expressions with the development of gastric cancer. The expressions of caveolin-1, E-cadherin and β-catenin were detected by biotin-streptavidin- peroxidase (SP) immunohistochemistry on 58 gastric cancer tissues, 40 precancerous gastric tissues and 42 chronic non-atrophic gastritis tissues. The correlation between the expressions of caveolin-1, E-cadherin and β-catenin, and the clinicopathologic parameters of gastric cancer was analyzed retrospectively. The positive rates of caveolin-1 and E-cadherin expressions in gastric carcinoma were significantly lower than precancerous gastric and chronic non-atrophic gastritis tissues (P<0.01). An abnormal rate of β-catenin expression in gastric carcinoma was higher than precancerous gastric and chronic non-atrophic gastritis tissues (P<0.01). Moreover, low expressions of caveolin-1, E-cadherin and β-catenin correlated with tumor size, depth of invasion, lymph node metastasis and TNM stage (P<0.05). The positive rates of caveolin-1 and E-cadherin expressions decreased (P<0.01), while an abnormal rate of β-catenin expression increased inversely, with the degree of atypical hyperplasia (P<0.01). Caveolin-1 expression correlated positively with E-cadherin (r=0.41, P<0.05). Caveolin-1 (r=-0.36, P<0.05) and E-cadherin (r=-0.45, P<0.05) expressions negatively correlated with abnormal β-catenin expression. These results suggested that dysregulated expressions of caveolin-1, E-cadherin and β-catenin correlated with the development of gastric cancer and its biological behavior.

Decreased blood riboflavin levels are correlated with defective expression ofRFT2gene in gastric cancer

To investigate the relationship between blood riboflavin levels and riboflavin transporter 2 (RFT2) gene expression in gastric carcinoma (GC) development. High-performance liquid chromatography was used to detect blood riboflavin levels in patients with GC. Real-time fluorogenic quantitative polymerase chain reaction and immunohistochemistry were used to analyze the expression of RFT2 mRNA and protein in samples from 60 GC patients consisting of both tumor and normal tissue. A significant decrease in the RFT2 mRNA levels was detected in GC samples compared with those in the normal mucous membrane (0.398 ± 0.149 vs 1.479 ± 0.587; P = 0.040). Tumors exhibited low RFT2 protein expression (75%, 16.7%, 8.3% and 0% for no RFT2 staining, weak staining, medium staining and strong staining, respectively), which was significantly lower than that in the normal mucous membrane (10%, 16.7%, 26.7% and 46.7% for no RFT2 staining, weak staining, medium staining and strong staining, respectively; P < 0.05). Tumors with low RFT2 expression were significantly associated with tumor stage and histological grade. Moreover, a significantly decrease in Uyghur patients was observed compared with Han patients. However, other parameters-gender, tumor location and lymph node metastasis-showed no significant relationship with RFT2 expression. Blood riboflavin levels were reverse correlated with development of GC (1.2000 ± 0.97569 ng/mL in high tumor stage patients vs 2.5980 ± 1.31129 ng/mL in low tumor stage patients; P < 0.05). A positive correlation of plasma riboflavin levels with defective expression of RFT2 protein was found in GC patients (χ² = 2.619; P = 0.019). Defective expression of RFT2 is associated with the development of GC and this may represent a mechanism underlying the decreased plasma riboflavin levels in GC.

Immunohistochemical expression of mTOR negatively correlates with PTEN expression in gastric carcinoma

The phosphoinositide-3 kinase (PI3K)-AKT-mammalian target of rapamycin (mTOR) pathway is a cellular pathway involved in cell growth, tumorigenesis and cell invasion which is frequently activated in various types of cancer. The downstream effector of the pathway is mTOR which is important in cellular growth and homeostasis and aberrant activation of mTOR has been reported in several types of cancer. The tumor suppressor gene phosphatase and tensin homolog (PTEN) is essential in this pathway for inhibiting tumor invasion and metastasis. However, the involvement of mTOR and PTEN in the progression of human gastric cancer remains to be identified. Immunohistochemical staining was performed to detect the expression of mTOR and PTEN in paraffin-embedded gastric tissue sections obtained from 33 patients with gastric cancer and 30 normal controls. The expressed mTOR was mainly distributed in the cytoplasm, while PTEN was mainly localized to the nucleus. By considering negative mTOR expression with positive PTEN expression as one group and negative PTEN expression with positive mTOR expression as the other, significant statistical differences were observed in various categories, including histological types and metastatic and clinical pathology stages, between the 2 groups (P<0.01 or 0.05). The results indicated that the expression levels of mTOR and PTEN were negatively correlated in the PI3K-AKT-mTOR signaling pathway. Combined detection of mTOR and PTEN expression may be used to evaluate the degree of malignancy in gastric cancer and may be a useful marker for the early diagnosis of gastric cancer.

Correlation between MMP-9/TIMP-1 imbalance and ZHX2 expression in gastric carcinoma

Correlation between MMP-9/TIMP-1 imbalance and ZHX2 expression in gastric carcinoma

Expression of FHL1 in gastric cancer tissue and its correlation with the invasion and metastasis of gastric cancer

This study was performed to analyze the expression of four and a half LIM domains 1 (FHL1) in gastric carcinoma tissue and its correlation with the clinicopathological characteristics of gastric cancer. In addition, the role of FHL1 in the invasion and metastasis of gastric cancer cells was investigated to provide an experimental basis for future treatments of gastric cancer. FHL1 mRNA and protein expression in gastric carcinoma and the adjacent normal gastric mucosa tissue were determined using RT-PCR and western blots. Correlations of FHL1 expression with the incidence, progression, and clinicopathological characteristics of gastric cancer were analyzed. Changes in the invasion and metastatic potential of MKN45 human gastric cancer cells were observed after the transient transfection with an eukaryotic expression vector containing full-length FHL1. Expression of FHL1 mRNA in gastric carcinoma tissue was significantly lower than that in the adjacent normal tissue (P < 0.05). FHL1 expression in gastric carcinoma tissue from patients who were positive for lymph node metastasis was significantly lower than those in patients who were negative for lymph node metastasis (P < 0.05). Lower FHL1 expression was correlated with lower degrees of differentiation, higher TNM stages, and greater invasive potential of the gastric cancer (P < 0.05). The FHL1 mRNA and protein expression patterns were similar in gastric cancer. FHL1 protein expression in gastric carcinoma tissue was significantly lower than that in the surrounding normal tissue (P < 0.05). FHL1 protein expression was significantly lower in gastric carcinoma tissue from patients who were positive for lymph node metastasis than that detected in patients with no lymph node metastasis (P < 0.05). Lower FHL1 protein expression was correlated with lower degrees of differentiation, higher TNM stages, and greater invasive potential in gastric cancer (P < 0.05). However, the expression of FHL1 was independent of the patient's gender, age, and tumor size (P > 0.05). Overexpression of FHL1 in the MKN45 human gastric cancer cell line using an eukaryotic expression vector resulted in a significant reduction in the invasiveness and metastatic ability of these cells as determined using the Transwell chamber invasion assay (P < 0.05). The decrease in or loss of FHL1 expression may be related to the incidence, progression, invasiveness, and metastatic potential of gastric cancer.

Dynamic expression of CEACAM7 in precursor lesions of gastric carcinoma and its prognostic value in combination with CEA

BackgroundThe significance of carcinoembryonic antigen-related cell adhesion molecule 7 (CEACAM7) expression in gastric carcinoma and precancerous lesions and its correlation with CEA expression has rarely been previously investigated.MethodsCEACAM7 and CEA expression was detected by immunohistochemistry in consecutive sections of 345 subjects with gastric carcinoma and precancerous lesions. Laser confocal analysis was performed to determine CEACAM7 and CEA localization. Correlation between CEACAM7 and CEA expression with clinicopathological parameters was statistically analyzed.ResultsCEACAM7 expression correlated with pathologic grading (P = 0.006), Lauren's classification (P = 0.023), and CEA expression (Spearman R = 0.605, P < 0.001) in gastric carcinoma. CEACAM7 co-localized with CEA predominantly in the cytoplasmic membrane of cancerous cells. CEA expression was correlated with lymph node metastasis (P = 0.031). CEACAM7 and CEA expression increased progressively from precursor lesions to gastric carcinomas. A combination of CEACAM7 and CEA expression was determined to be an independent predictor for patients with gastric carcinoma by multivariate analysis (P = 0.001).ConclusionsCEACAM7 expression correlates with tumor differentiation and CEA expression in gastric carcinoma. CEACAM7 and CEA expression may synergistically promote gastric carcinogenesis. Combined CEACAM7 and CEA expression analysis can be a useful postoperative predictor for patients with gastric carcinoma.

Prognostic significance of vascular endothelial growth factor expression in gastric carcinoma: a meta-analysis

The purpose of this study was to comprehensively and quantitatively summarize available evidences for the use of VEGF protein to evaluate the clinicopathological and prognostic role of VEGF expression in Asian patients with gastric cancer. Searches were applied to MEDLINE, EMBASE, and the Cochrane Library databases until June 2010, without language restrictions. A meta-analysis was performed to clarify the impact of VEGF expression on clinicopathological parameters or over survival (OS) in gastric cancer. Our combined results showed that VEGF expression in Asian patients with gastric cancer was significantly higher in the case-control studies (1,194 patients and 1,618 controls) (odds ratio [OR]=112.41, 95% confidence interval [CI]=64.12-197.06). All the analyses estimated favored a stronger link between the high VEGF expression and the poor 5-year overall survival (1,236 patients) (risk ratio [RR]=2.45, 95% CI=2.11-2.83, P=0.000). When stratifying the studies by the pathological variables, the depth of invasion (3,094 patients) (OR=1.95, 95% CI=1.40-2.71, P=0.000), lymph node metastasis (3,240 patients) (OR=1.82, 95% CI=1.29-2.57, P=0.001), distant metastasis (1,980 patients) (OR=2.76, 95% CI=1.22-6.25, P=0.015), vascular invasion (1,803 patients) (OR=2.61, 95% CI=2.09-3.27, P=0.000), and TNM stage (1,819 patients) (OR=1.92, 95% CI=1.57-2.36, P=0.000) provided significant prognostic information. Our results indicate that VEGF can potently predict the overall survival in Asian patients with gastric cancer. Importantly, VEGF may be converted from candidate to the routine clinical setting for clinicians to predict the outcome of single patient with gastric carcinoma.

Correlation between Expression of S100A4 and VEGF-C, and Lymph Node Metastasis and Prognosis in Gastric Carcinoma

This semiquantitative immunohistochemical study investigated the clinical significance of S100A4 and vascular endothelial growth factor C (VEGF-C) protein expression in gastric carcinoma. Correlations between S100A4 and VEGF-C immunoreactivity and clinicopathological characteristics were evaluated using 108 gastric carcinoma specimens and 20 specimens of tissue adjacent to gastric carcinoma. S100A4 and VEGF-C expression in carcinoma was higher than that in adjacent tissues. S100A4 expression was significantly related to tumour size and lymph node metastasis, whereas VEGF-C expression was associated with invasion depth, lymph node metastasis and tumour, node, metastasis (TNM) stage. A significant correlation was found between S100A4 and VEGF-C expression. Patients expressing S100A4 or VEGF-C showed no significant reduction in 5-year survival rate compared with those not expressing these proteins. Sex, age, tumour size, invasion depth, lymph node involvement, TNM stage, S100A4 expression and VEGF-C expression had a common effect on carcinoma prognosis but none was an independent prognostic factor.

P53 Expression in Gastric Dysplasia and carcinoma in Erbil City

Background: Alteration in the expression of p53 tumor-suppressor protein is an event that occurs frequently in human cancer, but the practical implications of this phenomenon are yet to be fully exploited.Objectives: to determine the value of p53 expression as a marker of tumor aggressiveness and the relationship between p53 over expression and clinico-pathologic variables in gastric adenocarcinoma.Material &amp; Methods: The expression of p53 was studied immunohisto-chemically in 10 cases with gastric dysplasia and 85 cases with gastric adenocarcinomas using formalin fixed, paraffin-embedded tissue samples. D07 a monoclonal antibody to p53 protein was used for the immunehistochemical analysis. The correlation between p53 expression in gastric dysplasia, gastric adenocarcinoma and clinicopathologic variables were analyzed.Results: P53 positivity was found in 48.2% of gastric carcinoma cases, and 40% of gastric dysplasia. There was a significant correlation between the rate of p53 over expression and tumor grade (p = 0.015), and also with carcinomas associated with lymph node involvement (p = 0034). There was no significant association between p53 protein expression and histological type (p = 0.171). The association between p53 protein expression and the depth of tumour invasion was found to be statistically insignificant.Conclusion: The results obtained suggest that P53 protein expression is a useful biomarker for the assessment of gastric adenocarcinoma aggressiveness. The preoperative assessment of p53 expression in gastric carcinomas can be helpful in identifying patients with higher grades and more advanced tumors. P53 immunoreactivity can predict lymph node status in patients with gastric carcinoma.

Expression of bone morphogenic protein-4 is inversely related to prevalence of lymph node metastasis in gastric adenocarcinoma

Bone morphogenic proteins (BMPs) are the largest subfamily of the transforming growth factor-β superfamily. Initially characterized as factors that induce bone and cartilage formation, BMPs have been found to be critical during mesoderm formation, organogenesis, and cellular differentiation. Bone morphogenic proteins are also known to modulate the morphologic alteration, adhesion, motility, and invasion of carcinoma cells derived from several organs. However, BMP-4 expression in gastric adenocarcinoma has not yet been clarified. We conducted the present study to define the clinical significance of BMP-4 expression in gastric carcinoma. Using immunohistochemistry, we investigated the expression of BMP-4 in normal mucosae and gastric adenocarcinoma samples from 64 patients with gastric carcinoma. The expression of BMP-4 was significantly higher in the adenocarcinoma than in the normal mucosae. Moreover, increased BMP-4 expression was associated with the presence of Helicobacter pylori infection. By contrast, the BMP-4 expression rate in gastric carcinoma was inversely related to the prevalence of lymph node metastasis and tumor invasiveness. The findings of this study suggest that BMP-4 expression may be a useful prognostic factor for predicting the outcome of patients with gastric carcinoma. Continued investigation to define the pathophysiologic mechanism underlying the role of BMP-4 in gastric carcinoma is warranted.

Abstract 2230: Quantitative expression of the hedgehog (Hh) signaling pathway in gastric carcinoma: A novel protocol to process RNA from paraffin-embedded tissue

Abstract Background: The Hh pathway is an evolutionally highly conserved signaling cascade important in gastroesophageal embryonic pattern formation and stem cell response to tissue damage. Its aberrant activation has also been implicated in gastric cancer formation and progression. Multiple therapeutics targeting the Hh pathway have been developed and in order to test their activity and pharmacodynamic responses, routine and consistent gene expression measurement from clinical samples must be available. Aims and Methods: The aim of this work was to measure levels of Hh pathway expression in both gastric tumors and normal gastric tissue, using a novel protocol of processing RNA from formalin fixed, paraffin-embedded tissue (FFPET). Total RNA was extracted from FFPET using a combination of Qiagen and RecoverAllTM kits according to the manufacturer's protocol(s). cDNA was synthesized using the iScriptTMcDNA Synthesis Kit and then a minimum of 20 ng RNA was pre-amplified using an Applied Biosystems pre-amplification cDNA kit. The primers for the Shh and Gli1genes were then added to the pre-amplification reaction and TaqManPreAmp master mix, amplified for 14 cycles on a thermal cycler and then real time PCR carried out for 40 cycles in duplicate. The relative quantification of the Shh and Gli1 genes were compared to the reference gene succinate dehydrogenase complex subunit A (SDHA) where the ratio for the change in expression for the selected gene = 2−ΔΔCT. Results: The levels of gene expression in gastric adenocarcinomas for Shh and Gli1 were respectively 3.89 ± 0.49 (mean ± SEM, N = 11) and 4.57 ± 1.46. For 8 non-tumor gastric specimens, the CT values for Shh ranged from 30.24 to 35.81 and for Gli1 from 31.21 to 34.81 while for SDHA the mean CT was 18.4 ± 1.87. Based on these CT values, no expression of Shh and Gli1 was present in these non-tumor gastric tissues. Conclusion: Hh pathway signaling can be quantitatively measured using as little as 20ng of RNA obtained from FFPET, making this methodology valuable in the clinic where tissue acquisition is often limited, as for gastric tumors. Using our methodology we found that there is increased Hh pathway expression in gastric carcinomas as compared to normal tissue, a finding consistent with previous reports in the literature. Supported by NO1-CM-07103-74 RECOVERY – ACTNOW Clinical Trial 8376 and NCI 2P30 CA16087-23, Tissue Acquisition and Banking Core. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2230. doi:10.1158/1538-7445.AM2011-2230

Latexin expression is downregulated in human gastric carcinomas and exhibits tumor suppressor potential

BackgroundLatexin, also known as endogenous carboxypeptidase inhibitor (CPI), has been found to inhibit mouse stem cell populations and lymphoma cell proliferation, demonstrating its potential role as a tumor suppressor. Our previous study also suggested a correlation between latexin expression and malignant transformation of immortalized human gastric epithelial cells. Here, we examined latexin expression in human gastric carcinomas and investigated the effect of differential latexin expression on proliferation of gastric cancer cells in vitro and in vivo.MethodsMonoclonal antibody against human latexin was prepared and immunohistochemical analysis was performed to detect latexin expression in 41 paired gastric carcinomas and adjacent normal control tissues. Human gastric cancer cells MGC803 (latexin negative) stably transfected with LXN gene and BGC823 cells (latexin positive) stably transfected with antisense LXN gene were established for anchorage-dependent colony formation assay and tumorigenesis assay in nude mice. Differentially expressed genes in response to exogeneous latexin expression were screened using microarray analysis and identified by RT-PCR. Bisulfite sequencing was performed to analyze the correlation of the methylation status of LXN promoter with latexin expression in cell lines.ResultsImmunohistochemical analysis showed significantly reduced latexin expression in gastric carcinomas (6/41, 14.6%) compared to control tissues (31/41, 75.6%) (P < 0.05). Overexpression of LXN gene in MGC803 cells inhibited colony formation and tumor growth in nude mice. Conversely, BGC823 cells transfected with antisense LXN gene exhibited enhanced tumor growth and colony formation. Additionally, several tumor related genes, including Maspin, WFDC1, SLPI, S100P, and PDGFRB, were shown to be differentially expressed in MGC803 cells in response to latexin expression. Differential expression of Maspin and S100P was also identified in BGC823 cells while latexin expression was downregulated. Further bisulfite sequencing of the LXN gene promoter indicated CpG hypermethylation was correlated with silencing of latexin expression in human cells.ConclusionsLatexin expression was reduced in human gastric cancers compared with their normal control tissues. The cellular and molecular evidences demonstrated the inhibitory effect of latexin in human gastric cancer cell growth and tumorigenicity. These results strongly suggest the possible involvement of latexin expression in tumor suppression.

Evaluation of HER2 Protein Expression in Gastric Carcinomas: Comparative Analysis of 1414 Cases of Whole-Tissue Sections and 595 Cases of Tissue Microarrays

Recent advances in molecular targeted therapy have identified HER2 as an important target for anti-cancer therapy in gastric cancer (GC). Although the clinical relevance and prognostic significance of HER2 in breast cancer has been well acknowledged, it remains controversial in GC. HER2 expression was investigated in two independent series of GC by immunohistochemical staining. One series corresponded to 1,414 cases of whole-tissue sections and the other corresponded to 595 cases of tissue microarrays (TMAs). Results were compared and correlated with clinicopathologic parameters. HER2-positivity was detected in 12.3% of whole-tissue sections and 17% of TMAs. Among samples scored 3+, 90.1% stained ≥50% of the tumor area, but only 40.9% in score 2+ cases stained ≥50% of the tumor area. In whole-tissue sections, HER2-positivity was correlated with age (P = 0.002), histological type (differentiated or intestinal, P < 0.001), lymphovascular invasion (P = 0.005), and lymph node metastasis (P = 0.009). In TMAs, HER2-positivity was correlated only with age (P = 0.003) and histological type (P < 0.001). Multivariate analyses of the differentiated GC subgroup revealed that HER2-positivity was an independent poor prognostic factor (P = 0.042). The cases with HER2-positive in ≥50% of the tumor area showed worse prognosis than those of <50% (P = 0.021). Despite discrepancies in the results from whole-tissue sections and TMAs, HER2 overexpression was positively correlated with aggressive biological behavior and was an independent poor prognostic factor for recurrence in differentiated GCs. Therefore, HER2-positive GCs should be considered for adjuvant trastuzumab therapy.

DNA methyltransferase 1 as a predictive biomarker and potential therapeutic target for chemotherapy in gastric cancer

PurposeDNA methylation contributes to carcinogenesis by mediating transcriptional regulation and chromatin remodelling, which may influence the effect of DNA-damaging drugs. We examined the prognostic and predictive impact of DNA methyltransferase (DNMT) 1 and 3b expression in gastric carcinomas (GC) treated by neoadjuvant chemotherapy. In vitro, DNMT1 expression and chemosensitivity were investigated for a functional relationship and the DNMT inhibitor decitabine (DAC) was tested as an alternative treatment option. Patients and methodsDNMT1/3b expression was analysed immunohistochemically in 127 pretherapeutic biopsies of neoadjuvant (platinum/5-fluorouracil)-treated GC patients and correlated with response and overall survival (OS). Short hairpin RNA technology was used to knockdown DNMT1 in the GC cell line, AGS. The chemosensitivity of GC cell lines to DAC alone and to DAC in combination with cisplatin was analysed by XTT or colony formation assays. ResultsHigh DNMT1 and DNMT3b expression was found in 105/127 (83%) and 79/127 (62%) carcinomas, respectively. Patients with low DNMT1 expression demonstrated a significantly better histopathological/clinical response (P=0.03/P=0.008) and OS (Plog-rank=0.001). In vitro, knockdown of DNMT1 caused an increased chemosensitivity towards cisplatin. Combined treatment with cisplatin and DAC showed a synergistic effect leading to increased cytotoxicity in the cisplatin-resistant cell line AGS. ConclusionLow DNMT1 expression defines a subgroup of GC patients with better outcomes following platinum/5FU-based neoadjuvant chemotherapy. In vitro data support a functional relationship between DNMT1 and cisplatin sensitivity. Besides its potential use as a predictive biomarker, DNMT1 may represent a promising target for alternative therapeutic strategies for a subset of GC patients.

HER2 gene status and the relationship with p21 protein expression in gastric cancer

We aimed to analysis the HER2 gene status and its relationship with p21 protein expression in gastric carcinoma. Fluorescence in situ hybridisation (FISH) and immunohistochemistry (IHC) techniques were used to detect HER2 gene status and p53 protein in 59 cases of gastric cancer. FISH detection of HER2 gene amplification rate was 16.9% (10/59), HER2 gene amplification in 49 cases without copy number gain and gene amplification were a total of 49.2% (29/59). HER2 protein expression was 42.4% (25/59), HER2 gene amplification rates in patients with +++, ++ HER2 protein expression were 3/3 and 5/8, while in patients with + HER2 protein expression, it was 2/14, there was significant difference (P 0.05). HER2 gene amplification rate and gene copy number had positively correlation with p21 protein expression, HER2 gene status and expression of p21 protein combined detection can provide a reference value in gastric cancer metastasis, patient’s condition development and prognosis, it also can guide clinical development of individual treatment.

Expressions of COX-2 and VEGF-C in gastric cancer: correlations with lymphangiogenesis and prognostic implications

BackgroundCyclooxygenase-2 (COX-2) has recently been considered to promote lymphangiogenesis by up-regulating vascular endothelial growth factor-C (VEGF-C) in breast and lung cancer. However, the impact of COX-2 on lymphangiogenesis of gastric cancer remains unclear. This study aims to test the expression of COX-2 and VEGF-C in human gastric cancer, and to analyze the correlation with lymphatic vessel density (LVD), clinicopathologic features and survival prognosis.MethodsUsing immunohistochemistry, COX-2, VEGF-C and level of LVD were analyzed in 56 R0-resected primary gastric adenocarcinomas, while paracancerous normal mucosal tissues were also collected as control from 25 concurrent patients. The relationships among COX-2 and VEGF-C expression, LVD, and clinicopathologic parameters were analyzed. The correlations of COX-2, VEGF-C and level of LVD with patient prognosis were also evaluated by univariate tests and multivariate Cox regression.ResultsThe expression rates of COX-2 and VEGF-C were 69.64% and 55.36%, respectively, in gastric carcinoma. Peritumoral LVD was significantly higher than that in both normal and intratumoral tissue (P < 0.05). It was significantly correlated with lymph node metastasis and invasion depth (P = 0.003, P = 0.05). VEGF-C was significantly associated with peritumoral LVD (r = 0.308, P = 0.021). However, COX-2 was not correlated with VEGF-C (r = 0.110, P = 0.419) or LVD (r = 0.042, P = 0.758). Univariate analysis showed that survival time was impaired by higher COX-2 expression and higher peritumoral LVD. Multivariate survival analysis showed that age, COX-2 expression and peritumoral LVD were independent prognostic factors.ConclusionsAlthough COX-2 expression was associated with survival time, it was not correlated with VEGF-C and peritumoral LVD. Our data did not show that overexpression of COX-2 promotes tumor lymphangiogenesis through an up-regulation of VEGF-C expression in gastric carcinoma. Age, COX-2 and peritumoral LVD were independent prognostic factors for human gastric carcinoma.

Assessment of ERBB2 and EGFR gene amplification and protein expression in gastric carcinoma by immunohistochemistry and fluorescence in situ hybridization

BackgroundThe goal of this study was to investigate ERBB2(HER2) and EGFR gene amplification and protein expression in gastric cancer. Fluorescence in situ hybridization (FISH) and immunohistochemistry were used to analyze ERBB2 and EGFR gene amplification and protein expression in 69 cases of gastric cancer.ResultsFISH analysis revealed that 20.3% of the cases exhibited ERBB2 gene amplification. Increases in ERBB2 copy number and gene amplification were present in 52.2% of the samples. Expression of the ERBB2 protein was observed in 42.0% of cases. FISH analysis detected EGFR gene amplification in 29.0% of samples. Increases in EGFR copy number and gene amplification occurred in 57.9% of samples, and EGFR protein expression was present in 52.2% of samples. Both ERBB2 and EGFR gene amplification were 3 cases (4.3%), but abnormalities in both ERBB2 and EGFR gene copy number were present 36.2% of samples. ERBB2 and EGFR gene amplification were significantly associated with the depth of tumor invasion (P < 0.05) and lymph node metastasis (P < 0.05), but not with sex, age, or histological type (P > 0.05).ConclusionsOur data indicated that ERBB2 and EGFR genetic abnormalities were associated with the prognosis of gastric cancer. Clinical assessment of ERBB2 and EGFR amplification may represent an important factor for the development of personalized treatment programs for gastic cancer.

Expression and prognostic significance of gastric‐specific annexin A10 in diffuse‐ and intestinal‐type gastric carcinoma

Annexin A10 (ANXA10) and its liver-specific short isoform (ANXA10S) had tissue-restricted expression. The downregulation of ANXA10S is correlated with tumor progression and poor prognosis in hepatocellular carcinoma. The aim of the present study was to validate the tissue distribution and explore the role of the ANXA10 protein expression in gastric carcinoma. We examined the ANXA10 protein expression in human and animal tissues and 356 resected primary gastric carcinomas, using specific mouse and rabbit polyclonal antibodies, by immunohistochemical staining. The ANXA10 protein is a nuclear protein specifically expressed in fetal and adult gastric mucosa and Brunner's gland across species, including humans, minipigs, woodchucks, and mice, and is commonly lost in gastric mucosa with intestinal metaplasia. The ANXA10 protein was expressed in 43.5% (155 cases) of gastric carcinomas; 74.2% (98/132) in the diffuse-type gastric carcinoma (DGC), 73.7% (28/38) in the mixed-type gastric carcinoma, and significantly lower in the intestinal-type gastric carcinoma (IGC) and indeterminate groups, 16.8% (28/167) and 5.3% (1/19), respectively (P<1×10(-8)). IGC with ANXA10 expression was correlated with a higher stage (P=0.049), particularly higher in stage IIIA/IIIB/IV IGC than lower-stage (IA/IB/II) tumors (P=0.005), but was not correlated with age, sex, and nodal status. In contrast, DGC with ANXA10 expression was associated with younger age, female patients, and importantly, lower tumor stage and lymph node metastasis (P=0.007, P=0.065, P=0.024, and P=0.0014, respectively). Moreover, DGC with ANXA10 expression had a better 5-year patient survival (P=0.0048), whereas IGC with ANXA10 expression had a lower 5-year survival (P=0.034). The ANXA10 protein expression is a novel marker of gastric differentiation, and is differentially expressed in IGC and DGC, with opposite prognostic significance.

Loss of BCL2L10 protein expression as prognostic predictor for poor clinical outcome in gastric carcinoma

BCL2L10 protein is an apoptosis-related member of the Bcl-2 protein family. The clinical significance of its expression in gastric carcinoma is poorly understood. The aim was to investigate BCL2L10 expression and its clinical and prognostic significance in gastric carcinoma patients. Immunohistochemistry, real-time polymerase chain reaction (PCR) and immunoblotting all revealed extensive loss of BCL2L10 expression in gastric cancer cells. The scaled BCL2L10 expression data was categorized into three groups (groups 0-2) to facilitate statistical analysis. A significant correlation was observed between the lower BCL2L10 expression group and shorter disease-free survival (P=1.956×10(-18)). Multivariate regression analysis showed that loss of BCL2L10 protein expression [P=4.883×10(-8), hazard ratio (HR)=0.252] is an independent prognostic predictor of gastric carcinoma. The receiver operator characteristic (ROC) curve showed that the area for BCL2L10 protein was 0.817 (P=8.331×10(-14)), indicating that loss of BCL2L10 protein expression is an excellent prognostic predictor of gastric carcinoma. Loss of BCL2L10 protein expression predicts poor clinical outcome in gastric carcinoma.