DAZAP1 promotes cancer progression and chemotherapy resistance by stabilizing PIN1 protein in gastric cancer.

Single cell and bulk RNA sequencing identifies tumor microenvironment subtypes and chemoresistance-related IGF1+ cancer-associated fibroblast in gastric cancer

Analysis of wntless (WLS) expression in gastric, ovarian, and breast cancers reveals a strong association with HER2 overexpression

Objective: To investigate the association between Thymidine phosphorylase(TYMP)genetic variation and clinical outcomes of postoperative gastric cancer (GC) patients received capecitabine based regimens. Methods: A total of 198 GC patients underwent surgical treatment and received capecitabine based adjuvant chemotherapy were included in this retrospective study. Peripheral blood and the postoperative tissue specimen of the GC patients were collected for the genotyping of polymorphism and TYMP mRNA expression, respectively. The correlation between polymorphism and clinical outcomes and safety of postoperative GC patients were analysed. Results: Located in the upstream, rs11479 was of clinical significance. The prevalence of rs11479 in TYMP among the GC patients were as follows: CC genotype 125 cases (63.13%), CT genotype 65 cases (32.83%), TT genotype 8 cases (4.04%), minor allele frequency of rs11479 is 0.20. The distribution of three genotypes were in accordance with Hardy-Weinberg Equilibrium (P=0.901). The analysis results of patients with different genotypes found that the 3-year disease free survival rate of the patients with CT/TT genotype and CC genotype were 73.97% and 65.60%, respectively, which was statistically significant (P=0.003). In terms of overall survival, the 3-year overall survival rate of the two genotypes were 83.56% and 72.80% (P=0.012), respectively. Adjusted in multivariate Cox regression analysis, CT/TT genotype was an independent favorable factor for disease free survival (OR=0.55, P=0.011). Safety analysis indicated that there was no significant association between genotypes and grade 2 adverse reaction. Additionally, of the 79 postoperative tissue specimens, the results showed that the expression of TYMP in cancer tissues of the patients with CT/TT genotypes were significantly higher than those of the wild type CC genotype patients (P<0.001). Conclusion: The polymorphism rs11479 of TYMP have favorable influence on the clinical outcomes of gastric cancer patients received capecitabine based adjuvant chemotherapy treatment through changing the mRNA expression of TYMP.

B-cell-specific Moloney murine leukemia virus integration site 1 (Bmi-1) and Raf kinase inhibitory protein (RKIP) are involved in cancer metastasis and chemotherapeutic resistance, respectively. In this study, we evaluated the association between Bmi-1 and RKIP and outcome of gastric cancer through clinical data analysis and in vitro experiments. Bmi-1 expression and RKIP expression were observed in 107 cases of gastric cancer through use of tissue microarray technology to identify their correlations with clinicopathological parameters, patient survival, and susceptibility to chemotherapy. The correlation was confirmed in gastric cancer cell lines, analyzed further by gene overexpression and silencing analysis, a cell invasion assay, and a chemosensitivity test. Positive expression of Bmi-1 was highly correlated with T classification and clinical stage. Diminished or lost expression of RKIP was significantly associated with T classification, lymph node metastasis, distant metastasis, and clinical stage. Bmi-1 is negatively and RKIP is positively related to patient survival. Positive expression of Bmi-1 and negative expression of RKIP are associated with poor patient survival and modest efficacy of postoperative chemotherapy. A meaningfully inverse association between Bmi-1 and RKIP was found in tissue microarray studies, and was verified further in gastric cancer cell lines. Moreover, gene overexpression and silencing analysis indicated that RKIP might be regulated by Bmi-1. Furthermore, the impacts of Bmi-1 on cell invasion and chemotherapy resistance were rescued by knockdown of RKIP. Our study implies that detection of Bmi-1 and RKIP is valuable in predicting patient survival and therapeutic response in gastric cancer, and the inverse association between Bmi-1 and RKIP reveals the potential molecular mechanisms underlying tumor metastasis and chemotherapy resistance.

Background and AimsThe average human life expectancy is increasing worldwide, thus the proportion of elderly gastric cancer patients is also increasing. In this case-control study, we investigated the clinical and oncologic outcomes of gastric cancer in patients over 80 years old.MethodsFrom January 2004 to December 2010, 291 patients aged over 80 years old (case group) were diagnosed and treated with gastric cancer at Asan Medical Center, Seoul, Korea. From the same period, 291 patients aged 18 to 80 years old were selected as the control group. The clinical findings and clinical outcomes of gastric cancer were retrospectively reviewed and compared between the two groups.ResultsThere were significant differences in the overall 5-year survival rate between the case and control groups (30.9% vs. 73.8%, respectively; P<0.001). In patients who received the curative treatment, overall 3- and 5-year survival rates showed 74.3% and 57.9% in case group and 91.6% and 86.5% in the control group. When analysis was confined to resectable elderly patients with a favorable performance, the curative resection group showed significantly better overall 3- and 5-year survival rates than the conservative treatment group (73.7% and 58.8% vs. 29.8% and 0%, respectively).ConclusionsAlthough elderly gastric cancer patients show an advanced stage at diagnosis and poor prognosis compared with non-elderly patients, elderly patients with good performance could benefit from curative resection. Thus, the clinical decision whether to undergo curative resection or conservative management should be made on an individualized basis.

30 Background: The average human life expectancy is increasing worldwide, thus proportion of elderly gastric cancer patients are also increasing. In this study, we investigated the clinical and oncologic outcomes of gastric cancer in patients over 80 years old through a case-control study. Methods: From January 2004 to December 2010, 291 patients aged over 81 years old (case group) were diagnosed and treated with gastric cancer at the Asan Medical Center. During the same period, 291 patients aged 18 to 80 years old were selected as control group. The clinical findings, histopathological parameters, and clinical outcomes of gastric cancer were reviewed retrospectively and compared between the two groups. Results: There were significant differences in overall 5-year survival rate between the two groups (30.9% vs 73.8%, P&lt; 0.001). When analysis was confined to resectable elderly patients with favorable performance of American Society of Anesthesiologists (ASA) score 1 or 2, curative resection group showed significantly better overall 3- and 5-year survival rate than the conservative treatment group (73.7% and 58.8% vs 29.8% and 0%, respectively). In multivariate analysis, lower BMI and advanced TNM stage were found to be independent prognostic predictors for poorer survival. ASA score showed borderline significance for predictors for poorer survival (P=0.087). Conclusions: Although elderly patients showed advanced stage at diagnosis and poor prognosis compared to non-elderly patients, elderly patients with good performance could benefit from curative resection of gastric cancer, thus the clinical decision whether to undergo curative resection or conservative management should be made on individualized approach.

Quantitative assessment of gene methylation and their impact on clinical outcome in gastric cancer

Gastric cancer (GC) is the third most common cause of cancer‑related death in the world. Annexin A10 (ANXA10), a member of the Annexin family, is a calcium‑/phospholipid‑binding protein; however, little is known concerning its functions. It is still unclear what molecule is involved in the induction of ANXA10. In the present study, we performed immunohistochemistry to evaluate the expression of ANXA10, pancreatic and duodenal homeobox‑1 (PDX1) and mucin phenotype markers in 130GC samples. ANXA10 was detected in 63(48%) of the 130GC cases and loss of ANXA10 was significantly correlated with disease progression and poor clinical outcomes in GC. PDX1 was significantly correlated with ANXA10 in GC cases and cell lines. Although PDX1 was not significantly correlated with the GC cases with any of the mucin phenotypes, ANXA10 was preferentially detected in the GC cases with the gastric mucin phenotype. As a further investigation, we generated organoids derived from human GC and identified the duplication of the mucin phenotypes of GC by immunohistochemistry. The repression effect on cell growth that was observed in the ANXA10‑knockdown cell lines was also clearly observed in the human gastric organoids. We demonstrated that the expression of ANXA10 was correlated with the gastric mucin phenotype and ANXA10 was involved in the induction of PDX1 expression in GC. We also provided evidence that GC organoids represent a powerful tool for scrutinizing the biology of GC, especially with regard to the mucin phenotype.

Dysregulation of the c-MET signaling pathway results from various molecular mechanisms including mutation, amplification, and overexpression. Overexpression and amplification of c-MET have been correlated with poor clinical outcome in gastric cancer, whereas the associations between c-MET polymorphisms and prognosis have not been well defined. We examined the prognostic impact of functional polymorphisms of the MET gene on clinical outcome in gastric cancer. Candidate polymorphisms of the MET gene were analyzed by PCR-based direct sequencing for the associations with clinical outcome across three independent cohorts, including 161 Japanese, 101 US, and 63 Austrian patients, with locoregional gastric cancer, treated with surgery. The univariable analysis showed that patients with any G (A/G or G/G genotype) allele of MET rs40239 had significantly longer disease-free survival and overall survival compared with those with the AA genotype in the Japanese cohort [hazard ratio (HR): 0.43, P=0.001, and HR: 0.47, P=0.006, respectively]; this remained significant upon multivariable analysis adjusted for age, sex, stage, and type of adjuvant therapy (HR: 0.48; P=0.009, HR: 0.50; P=0.017, respectively). However, there was no significant association of the polymorphism with clinical outcome in the US and Austrian cohorts. When stratified by sex in the Japanese cohort, male individuals, but not female individuals, with the G allele maintained a clinical outcome benefit in both univariable and multivariable analyses. MET rs40239 may serve as a prognostic biomarker in locoregional gastric cancer. These data also suggest that genetic variants of c-MET may show sex-related differences in the impact on clinical outcome.

BackgroundGastric cancer (GC) is a common malignant disease and microRNAs (miRNAs) have been shown to play important roles in GC tumorigenesis. As the clinical outcome of GC is closely correlated with the clinical stage at the time of diagnosis, early detection and prevention are crucial. This study was designed to evaluate the expression level of plasma miR-23b in patients with GC and investigate the relationship between plasma miR-23b expression level and the prognosis of GC.Material/MethodsWe recruited 138 patients diagnosed with GC and 50 healthy volunteers. Quantitative real-time PCR (qRT-PCR) was performed to evaluate the expression level of plasma miR-23b in all participants. The association between miR-23b expression and clinicopathological factors as well as survival rates was analyzed. Receiver operator curve (ROC) analysis was carried out to evaluate the diagnostic performance of plasma miR-23b for GC.Univariate and multivariate Cox regression analyses were conducted to determine whether plasma miR-23b was an independent predictor of survival.ResultsThe expression levels of miR-23b were upregulated in plasma samples from GC patients (P<0.01) and were significantly associated with T stage, distant metastasis, and differentiation. Significantly shorter 5-year overall survival (OS) and disease-free survival (DFS) were observed in patients with higher expression of the miR-23b (P<0.01). The area under the curve (AUC) of high expression of plasma miR-23b to diagnose GC was 0.80 (95% CI: 0.74–0.86, P<0.001). Multivariate analysis revealed that enhanced expression of plasma miR-23b was an independent predictor of OS (P=0.015) and DFS (P=0.004).ConclusionsOur results indicated that plasma miR-23b was overexpressed in GC patients and high plasma miR-23b expression was associated with poor clinical outcome. Thus, plasma miR-23b may serve as a potential diagnostic biomarker and therapeutic target for GC.

This study aims to investigate the expression and significance of GOLPH3 in human gastric cancer progression and prognosis. Using immunohistochemistry (IHC) and real-time reverse transcriptase polymerase chain reaction assay, we identified abnormally elevated expression of GOLPH3 in gastric cancer tissues compared to paired normal stomach mucosa tissues in 40 patients. In addition, the enzyme-linked immunosorbent assay (ELISA) was used to quantify serum GOLPH3 concentrations in the same 40 gastric cancer patients and 40 healthy individuals. ELISA revealed significantly higher serum concentrations of GOLPH3 in gastric cancer patients compared to healthy individuals (p = 0.002). In order to investigate the correlations between GOLPH3 and the clinicopathological features of gastric cancer, the expression of GOLPH3 in 123 gastric cancer patients were detected by IHC, and the results showed that overexpression of GOLPH3 was associated with the size of the tumor (p = 0.013), histological grade (p = 0.002), depth of invasion (p < 0.001), lymph node metastasis (p < 0.001), distant metastasis (p = 0.018), and TNM stage (p < 0.001). Kaplan-Meier survival analysis showed that high GOLPH3 expression exhibited a significant correlation with poor prognosis for gastric cancer patients. Further, Cox multivariate analysis indicated that GOLPH3 expression level was an independent prognostic factor for patients after radical resection. In conclusion, the overexpression of GOLPH3 is closely related to the progression of gastric cancer and might be regarded as an independent predictor of poor prognosis for gastric cancer.

Background: Tumor-associated macrophages (TAMs) are prominent immune cells infiltrating in solid tumors with phenotypic and functional heterogeneity. However, the clinical significance of heterogeneous subtypes of TAMs in gastric cancer still remains obscure. Here we aimed to explore the clinical significance of TAMs expressing dendritic cellspecific intercellular adhesion molecule-3-grabbing non-integrin (DCSIGN) and its potential influence on immune contexture in gastric cancer. Methods: We selected 453 formalin-fixed and paraffin-embedded tissue microarray samples and 41 fresh tissue specimens of patients with gastric cancer from Zhongshan Hospital. The association of DC-SIGN+ macrophages with clinicopathological parameters, overall survival (OS) and responsiveness to fluorouracil-based adjuvant chemotherapy (ACT) was inspected by immunohistochemistry (IHC) evaluation. IHC and flow cytometry were applied to characterize the immune contexture in gastric cancer. Findings: We demonstrated that high intratumoral DC-SIGN+ macrophages infiltration predicted poor OS and inferior therapeutic responsiveness to ACT in patients with gastric cancer. DC-SIGN+ macrophages orchestrated an immunoevasive contexture in the tumor microenvironment with more protumor Foxp3+ regulatory T cells (Tregs) and dysfunctional CD8+ T cells yet fewer anti-tumor CD138+ plasma cells. Furthermore, dysfunctional CD8+ T cells in patients with high DC-SIGN+ macrophages infiltration exhibited with decreased interferon-γ (IFN-γ), granzyme B (GZMB) and perforin-1 (PRF1) expression yet elevated programmed cell-death protein 1 (PD-1) and cytotoxic T-lymphocyte-associated protein 4 (CTLA-4) expression. Moreover, CD8+ T cells cytotoxicity was impeded by DC-SIGN+ macrophages derived latency-associated peptide (LAP). Interpretation: Our results suggest that DC-SIGN+ macrophages could be applied as an independent predictive marker and a potential immunotherapeutic target for gastric cancer. Funding Statement: This study was funded by grants from National Natural Science Foundation of China (31470794, 81471621, 81472227, 81501999, 81671628, 81672324, 31770851, 81871306, 81871926, 81871930), Shanghai Municipal Natural Science Foundation (18ZR1432900), and Shanghai Sailing Program (17YF1402200, 18YF1404600, 19YF1407500). Declaration of Interests: The authors declare no conflict of interest. Ethics Approval Statement: All patients in this study were provided with written informed consent, and this study was approved by the Clinical Research Ethics Committee of Zhongshan Hospital, Fudan Univeristy.

Background:SRY-related HMG box-12, which is associated with the prognosis of cancer, has been frequently described. However, both SRY-related HMG box-12 expression and its relationship with clinicopathological variables and patient survival have not been defined in gastric cancer. The aim of our study was to examine the prognostic value of SRY-related HMG box-12 expression in patients with gastric cancer.Methods:In this study, we determined SRY-related HMG box-12 expression in 79 primary gastric cancer tissues and 79 matched adjacent nontumor tissues by immunohistochemistry and then calculated the survival rate using the Kaplan-Meier method. Cox proportional hazard regression model was used to analyze predictors of gastric cancer. Western blot and quantitative real-time polymerase chain reaction were used to investigate the difference in SRY-related HMG box-12 expression between normal gastric epithelial cells and gastric cancer cells at the protein level and RNA level, respectively.Results:SRY-related HMG box-12 was downregulated in gastric cancer tissues. Low SRY-related HMG box-12 expression was significantly associated not only with lymph node metastasis (P = .027) and TNM stage (P = .021) but also with disease-specific survival in patients with gastric cancer. Multivariate analysis demonstrated TNM stage was an independent factor predicting poor survival (P = .034).Conclusions:Low SRY-related HMG box-12 expression is associated with poor clinical outcomes in gastric cancer.

The tumor suppressor role of tissue factor pathway inhibitor 2 (TFPI-2) has been reported in various tumors. The present study aimed to improve the understanding of the oncogenic properties of TFPI-2 in gastric cancer. Relative expression of TFPI-2 was determined by a real-time polymerase chain reaction (PCR) and western blotting, respectively. Cell viability was measured via a cell counting kit-8 assay and proliferation was evaluated by a colony formation assay. Cell apoptosis was assessed with a caspase-3 activity kit and invasion was evaluated by a transwell chamber assay. The methylation level of TFPI-2 promoter was assayed by methylation-specific PCR. The regulatory effect of miR-27a-3p on TFPI-2 was analyzed with a luciferase reporter assay. The direct association between miR-27a-3p and TFPI-2 was shown by biotin-labelling pulldown. TFPI-2 was down-regulated in gastric cancer, which associated with an unfavorable prognosis clinically. Ectopic introduction of TFPI-2 greatly compromised cell viability, colony formation and invasive capacity, and also induced cell apoptosis simultaneously. The promoter region of TFPI-2 was extensively methylated in gastric cancer tissues compared to normal tissues, suggesting the epigenetic inhibition of TFPI-2 expression. We further identified that TFPI-2 functioned as sponge RNA against miR-27a-3p. Most importantly, miR-27a-3p-specific inhibitor significantly exerted a tumor suppressor function akin to TFPI-2 itself, and the anti-tumoral activities were completely abolished by TFPI-2 knockdown. We found that the epigenetically suppressed TFPI-2 compromised sponging effects with respect to miR-27a-3p in gastric cancer, which consequently and mechanistically contributed to the tumor biology of gastric cancer.

This study aims to investigate the expression and significance of glucose-6-phosphate dehydrogenase (G6PD) in human gastric cancer progression and prognosis. Using immunohistochemistry and real-time RT-PCR assay, we identified abnormally elevated expression of G6PD in gastric cancer tissues compared to paired normal stomach mucosa tissues in 24 patients (p < 0.05). In order to investigate the correlations between G6PD and the clinicopathological features of gastric cancer, the expression of G6PD in 167 patients with gastric cancer were detected by immunohistochemistry, and the results showed that overexpression of G6PD was associated with the size of tumor (p = 0.039), depth of invasion (p = 0.039), lymph node metastasis (p = 0.044), distant metastasis (p = 0.003), TNM stage (p = 0.030), and survival rate (p = 0.010). Further, Cox multivariates analysis indicated that G6PD expression level was an independent prognostic factor for patients after radical resection (p = 0.013). In conclusion, overexpression of G6PD is closely related to progression of gastric cancer, and might be regarded as an independent predictor of poor prognosis for gastric cancer.