The association of immune cell infiltration and prognostic value of tertiary lymphoid structures in gastric cancer.

Tertiary lymphoid structures (TLS) are lymphoid aggregates in tumor tissues and their potential significance in clinical applications has not been fully elucidated in gastric cancer. We evaluated TLS and tumor-infiltrating immune cells using H&E and immunohistochemistry staining in the recruited patients with gastric cancer. The prognostic value of TLS was evaluated by Kaplan-Meier analysis and further validated using gene expression profiling. The alterations in gene mutation, copy number variance, and DNA methylation across the TLS signature subtypes were analyzed based on the Cancer Genome Atlas cohort. High TLS density was associated with improved overall survival and disease-free survival. A combination of TLS density and TNM stage obtained higher prognostic accuracy than the TNM stage alone. Tumors with high TLS density showed significantly higher infiltration of CD3+, CD8+, and CD20+ cells but lower infiltration of CD68+ cells. Transcriptomics analysis demonstrated that high TLS signature status was positively associated with the activation of inflammation-related and immune-related pathways. Multi-omics data showed a distinct landscape of somatic mutations, copy number variants, and DNA methylation across TLS signature subtypes. Our results indicated that TLS might link with enhanced immune responses, and represent an independent and beneficial predictor of resected gastric cancer. Multi-omics analysis further revealed key tumor-associated molecular alterations across TLS signature subtypes, which might help explore the potential mechanism of the interaction between TLS formation and cancer cells.

The Anti-Tumor Effect of Nab-Paclitaxel Proven by Patient-Derived Organoids.

BACKGROUND: Nab-paclitaxel has been widely used in treating breast cancer and pancreatic patients for its low toxicity and high efficiency. However, its role in gastric cancer (GC) remains ambiguous. The aim of our study was to test the anti-tumor activity of nab-paclitaxel using GC patient-derived organoids. METHODS: By using the organoid culture system, we describe the establishment of human gastric cancer organoid lines from surgical samples of three patients with gastric cancer. The consistency of these organoids with original cancer tissues was evaluated by histopathological examination. The characteristics of the cancer organoids were tested using immunofluorescence (IF) staining. Using organoids, the anti-tumor efficiencies of nab-paclitaxel, 5-Fu and epirubicin were compared by CCK8 assay and Annexin V-FITC/PI staining. RESULTS: Three organoids were successfully established and passaged. The morphology of the established GC organoids was consistent with original cancer tissues. The IC50 of nab-paclitaxel was 3.68 µmol/L in hGCO1, 2.41 µmol/L in hGCO2 and 2.91 µmol/L in hGCO3, which was significantly lower than those of 5-FU (72.99 µmol/L in hGCO1, 28.32 µmol/L in hGCO2 and 2.91 µmol/L in hGCO3) and epirubicin (25.85µmol/L in hGCO1, 15.15 µmol/L in hGCO2 and 7.60 µmol/L in hGCO3). When each organoid lines were treated with nab-paclitaxel for increasing period of time, the percentage of the apoptotic cells in each organoid increased accordingly. CONCLUSION: Nab-paclitaxel showed strong anti-tumor activity and had the potential to become front-line drug for treating GC patients. Gastric cancer organoid may be a good tool to predict in vivo response to drugs.

Immune-infiltration based signature as a novel prognostic biomarker in gastrointestinal stromal tumour.

BACKGROUND: Accumulating evidence indicates that tumour-infiltrating lymphocytes (TILs) are the primary determinant of survival outcomes in various tumours. Thus, we sought to investigate the TIL distribution and density in gastrointestinal stromal tumours (GISTs) and to develop an immune infiltration (II)-based signature to predict prognosis. METHODS: The expression of 8 immune features in the tumour centre (TC) and tumour margin (TM) and PD-L1 in 435 GIST patients was investigated by immunohistochemistry. Then, a 4-feature-based II-GIST signature integrating the CD3+ TC, CD3+ TM, CD8+ TM and CD45RO+ TM parameters was developed using a LASSO Cox regression model in the training cohort and was validated in two separate validation cohorts. FINDINGS: High CD3+ TC, CD3+ TM, CD8+ TC, CD8+ TM, CD45RO+ TM, NKp46+ TM and CD20+ TM correlated with improved survival. Patients with high II-GIST scores have better RFS and OS outcomes than those with low II-GIST scores. Multivariable analyses demonstrated that the II-GIST signature is an independent prognostic factor. The receiver operating characteristic (ROC) curve demonstrated that the prognostic accuracy of the II-GIST signature is superior to that of the NIH risk criteria. Further analysis showed that moderate- and high-risk GIST patients with high II-GIST scores could gain survival benefits from adjuvant imatinib therapy. INTERPRETATION: The novel II-GIST signature accurately predicted the survival outcomes of GIST patients. In addition, the II-GIST signature was a useful predictor of survival benefit from imatinib therapy amongst moderate- and high-risk patients with GIST. FUNDING: This project was supported by National Natural Science Foundation of China (81702325), Natural Science Foundation of Guangdong Province (2017A030310565), and 3&3 Project of the First Affiliated Hospital of Sun Yat-sen University.

The prognostic roles of platelet-to-lymphocyte ratio and neutrophil-to-lymphocyte ratio in gastrointestinal stromal tumours: a meta-analysis.

BACKGROUND: The platelet-to-lymphocyte ratio (PLR) and neutrophil-to-lymphocyte ratio (NLR) have been found to be associated with prognosis in several solid tumours. However, the prognostic roles of PLR and NLR in gastrointestinal stromal tumours (GISTs) remain controversial. The aim of this meta-analysis was to assess the prognostic roles of PLR and NLR in GISTs. METHODS: We searched MEDLINE, EMBASE and the Cochrane Library for relevant articles. A systematic review was performed to calculate pooled hazard ratios (HRs) for disease-free survival (DFS) and overall survival (OS) by fixed-effects/random-effects models. RESULTS: Fourteen studies containing 3,151 subjects were finally enrolled in this meta-analysis. Eight studies including 2,560 patients investigated the prognostic effect of PLR, and thirteen studies with 2,751 subjects explored the prognostic effect of NLR. Both elevated PLR (HR: 1.29, 95% CI: 1.10-1.52, P=0.002) and NLR (HR: 1.37, 95% CI: 1.15-1.63, P=0.0005) were significantly associated with decreased DFS. The pooled HR for PLR was not significantly different from that for NLR. High PLR and NLR correlated with increased tumour sizes, more advanced tumour stages and mitotic index (>5/50 HPF). In addition, elevated PLR was related to adjuvant tyrosine kinase inhibitor (TKI) therapy. CONCLUSIONS: Elevated preoperative PLR and NLR are associated with poor outcomes in patients with GISTs.

Predictive ability of prognostic nutritional index in surgically resected gastrointestinal stromal tumors: a propensity score matching analysis.

BACKGROUND: Recent findings have shown that inflammation indices are associated with prognosis in various malignancies. However, the usefulness of inflammation indices including platelet-to-lymphocyte ratio, neutrophil-to-lymphocyte ratio and prognostic nutritional index for gastrointestinal stromal tumors (GISTs) remains controversial. METHODS: We retrospectively reviewed 340 primary localized GIST patients who had received surgical resection between 2005 and 2015 to investigate the effect of platelet-to-lymphocyte ratio, neutrophil-to-lymphocyte ratio and prognostic nutritional index on prognosis. 206 patients were selected by propensity score matching to control selection biases. RESULTS: Kaplan-Meier analysis and the log rank test demonstrated that high prognostic nutritional index (≥43.9) was significantly correlated with better recurrence-free survival (RFS) (P<0.001). Among the three inflammatory indices, only preoperative high prognostic nutritional index was an independent prognostic factor for survival [hazard ratio (HR) 0.509; 95% confidence interval (CI) 0.266-0.872; P = 0.031] in multivariate analysis. After propensity score matching, elevated prognostic nutritional index was still a predictor for RFS (HR = 0.498; 95% CI 0.253-0.971; P = 0.042) in the multivariate analyses. In addition, prognostic nutritional index was a significant prognostic factor for GISTs within the National Institutes of Health (NIH) high and very low/low-risk categories. Incorporation prognostic nutritional index into the NIH risk criteria improved the prognostic stratification (c-index, 0.725 vs. 0.686, p = 0.039). CONCLUSIONS: High prognostic nutritional index is a predictor of improved survival for surgically resected GISTs and incorporation prognostic nutritional index into NIH risk criteria improves the predictive accuracy.

Distinct Prognosis of High Versus Mid/Low Rectal Cancer: a Propensity Score-Matched Cohort Study.

BACKGROUND: Rectal cancers have long been treated as a single-entity disease; however, whether the prognosis of high rectal cancer (inferior margin located 10.1 to 15.0 cm from the anal verge) differs from that of mid/low rectal cancer (0 to 10.0 cm) remains disputed. METHODS: Patients with stages I-III rectal adenocarcinomas undergoing curative-intent surgery were enrolled between 2007 and 2013 in this retrospective analysis. Exclusion criteria were neoadjuvant therapy or concurrent cancers. Propensity score matching and Cox regression analysis were performed to compare a 5-year overall and cancer-specific survival between patients with high and mid/low rectal cancer. RESULTS: Of 613 patients who met the inclusion criteria, 199 (32.5%) and 414 (67.5%) had high and mid/low rectal cancer, respectively. After propensity score matching (187 cases for each group), the high group showed a better overall survival (70.9 vs. 56.9%, p = 0.042) and cancer-specific survival (77.4 vs. 60.3%, p = 0.028) at 5 years compared with the mid/low group with stage III disease. However, high rectal cancer did not demonstrate prognostic superiority in stages I-II disease. Multivariate analysis identified high tumor location as an independent prognostic factor for cancer-specific survival (hazards ratio = 0.422, 95% confidence interval 0.226-0.786, p = 0.007) and overall survival (hazards ratio = 0.613, 95% confidence interval 0.379-0.991, p = 0.046). CONCLUSIONS: Patients with stage III high rectal adenocarcinoma demonstrated better overall and cancer-specific survival than those with mid/low type, and tumor location was an independent prognostic factor for patients with rectal carcinomas.

CXCL1 expression is correlated with Snail expression and affects the prognosis of patients with gastric cancer.

Gastric cancer (GC) continues to result in a poor survival rate and prognostic biomarkers for the disease are lacking. Chemokine (C-X-C motif) ligand (CXCL1) expression plays a critical role in tumor metastasis, and Snail promotes epithelial-mesenchymal transition (EMT) to promote metastasis. Therefore, the present study aimed to investigate the correlation between CXCL1 and Snail expression and the effect of CXCL1 expression on the survival of patients with GC. CXCL1 and Snail expression in paraffin-embedded tissue sections from 127 patients with GC were each assessed by immunohistochemistry. Cox regression and Kaplan-Meier analyses were performed to evaluate the prognostic significance of CXCL1 and Snail. Evaluation of the association between CXCL1 and Snail expression and clinical characteristics was based on the χ2 test. Spearman's rank correlation coefficient and Fisher's exact test were used to explore the association between CXCL1 and Snail expression in GC tissues. CXCL1 was found to be significantly associated with tumor invasion (P=0.003), tumor-node-metastasis (TNM) staging (P=0.001), tumor size (P=0.013) and lymph node metastasis (P=0.022) in GC. Snail overexpression was also significantly associated with tumor invasion (P=0.001), TNM staging (P=0.005), tumor size (P=0.026), lymph node metastases (P=0.014) and perineural invasion (P=0.009). CXCL1 and Snail expression were independent factors for a worse overall survival rate, as determined by multivariate analysis (P=0.011 and P=0.018; respectively). The combined expression of CXCL1 and Snail resulted in a worse prognosis compared with the other three groups (P=0.005). Furthermore, there was a significantly positive correlation between CXCL1 and Snail expression in GC (r=0.431; P<0.001). The expression of CXCL1 is significantly associated with Snail expression and may be used as a predictive co-biomarker for patient prognosis and tumor aggressiveness in GC. CXCL1 may promote GC metastasis by regulating EMT.

CXCL1 promotes tumor growth through VEGF pathway activation and is associated with inferior survival in gastric cancer.

The chemokine (C-X-C motif) ligand 1 (CXCL1) regulates tumor-stromal interactions and tumor invasion. However, the precise role of CXCL1 on gastric tumor growth and patient survival remains unclear. In the current study, protein expressions of CXCL1, vascular endothelial growth factor (VEGF) and phospho-signal transducer and activator of transcription 3 (p-STAT3) in primary tumor tissues from 98 gastric cancer patients were measured by immunohistochemistry (IHC). CXCL1 overexpressed cell lines were constructed using Lipofectamine 2000 reagent or lentiviral vectors. Effects of CXCL1 on VEGF expression and local tumor growth were evaluated in vitro and in vivo. CXCL1 was positively expressed in 41.4% of patients and correlated with VEGF and p-STAT3 expression. Higher CXCL1 expression was associated with advanced tumor stage and poorer prognosis. In vitro studies in AGS and SGC-7901 cells revealed that CXCL1 increased cell migration but had little effect on cell proliferation. CXCL1 activated VEGF signaling in gastric cancer (GC) cells, which was inhibited by STAT3 or chemokine (C-X-C motif) receptor 2 (CXCR2) blockade. CXCL1 also increased p-STAT3 expression in GC cells. In vivo, CXCL1 increased xenograft local tumor growth, phospho-Janus kinase 2 (p-JAK2), p-STAT3 levels, VEGF expression and microvessel density. These results suggested that CXCL1 increased local tumor growth through activation of VEGF signaling which may have mechanistic implications for the observed inferior GC survival. The CXCL1/CXCR2 pathway might be potent to improve anti-angiogenic therapy for gastric cancer.

Impact of pre-existing type-2 diabetes on patient outcomes after radical resection for gastric cancer: a retrospective cohort study.

BACKGROUND: The aim of this study was to evaluate the impact of pre-existing type-2 diabetes on postoperative recovery and prognosis in gastric cancer (GC) patients who underwent radical gastrectomy. RESEARCH DESIGN AND METHODS: From June 2001 to June 2011, a total of 1,014 eligible patients were enrolled. Among them, 67 patients were diagnosed with type-2 diabetes. The clinicopathologic features and prognostic data were compared between patients with type-2 diabetes (the DM group) and without diabetes (the non-DM group). RESULTS: Median survival was 68.3 months. The 5-year overall survival in the DM group was similar to that in the non-DM group (52.1 vs. 53.0 %, p = 0.411). Propensity score matching analysis demonstrated that the hazard ratio of death in the DM group was 1.191 (95 % confidential index 0.693-2.072; p = 0.531) compared to the-non DM group. Incidence of postoperative complications was higher in the DM group than in the non-DM group (17.9 vs. 8.1 %, p = 0.006). The DM remission rate was 46 % among patients who received Roux-en-Y reconstruction, and 13 % among patients who received Billroth II anastomosis (p = 0.009). The 5-year overall survival rate was 62.1 % for patients with cured or improved DM and 23.4 % for patients with worse or same DM status (p = 0.003). CONCLUSION: Type-2 diabetes can be cured by radical gastrectomy plus Roux-en-Y reconstruction in some GC patients. Pre-existing diabetes is associated with increased postoperative complications and decreased survival when it becomes worse after curative dissection for GC.

[Meta-analysis of adjuvant chemotherapy on prognosis for gastric cancer patients after D2 dissection].

OBJECTIVE: To evaluate efficacy of adjuvant chemotherapy after D2 dissection on survival for patients with gastric cancer. METHODS: Randomized clinical trials (RCT) that compared adjuvant chemotherapy after D2 dissection with D2 dissection alone for gastric cancer were searched with Pubmed, Cochrane, Embase and CBM databases. Eligible trials published between 1990 and 2012 were included in the study. The quality of RCTs was assessed by the Jadad scale. Data synthesis and statistical analysis were performed by RevMan 5.1 software. RESULT: Eight RCTs with 3633 patients were included in this study. Among them, 1824 patients received adjuvant chemotherapy and 1809 patients didn't. Adjuvant chemotherapy was associated with a significant benefit in terms of overall survival (RR = 0.76, 95% CI: 0.69-0.84), disease free survival (RR = 0.72, 95%CI: 0.66-0.80) and recurrence rate (RR = 0.69, 95% CI: 0.62-0.77). CONCLUSION: Adjuvant chemotherapy was associated with survival benefit for gastric cancer after D2 dissection.

Efficacy of adjuvant XELOX and FOLFOX6 chemotherapy after D2 dissection for gastric cancer.

AIM: To compare the efficacy of capecitabine and oxaliplatin (XELOX) with 5-fluorouracil, folinic acid and oxaliplatin (FOLFOX6) in gastric cancer patients after D2 dissection. METHODS: Between May 2004 and June 2010, patients in our gastric cancer database who underwent D2 dissection for gastric cancer at the First Affiliated Hospital of Sun Yat-Sen University were retrospectively analyzed. A total of 896 patients were enrolled into this study according to the established inclusion and exclusion criteria. Of these patients, 214 received the XELOX regimen, 48 received FOLFOX6 therapy and 634 patients underwent surgery only without chemotherapy. Overall survival was compared among the three groups using Cox regression and propensity score matched-pair analyses. RESULTS: Patients in the XELOX and FOLFOX6 groups were younger at the time of treatment (median age 55.2 years; 51.2 years vs 58.9 years), had more undifferentiated tumors (70.1%; 70.8% vs 61.4%), and more lymph node metastases (80.8%; 83.3% vs 57.7%), respectively. Overall 5-year survival was 57.3% in the XELOX group which was higher than that (47.5%) in the surgery only group (P = 0.062) and that (34.5%) in the FOLFOX6 group (P = 0.022). Multivariate analysis showed that XELOX therapy was an independent prognostic factor (hazard ratio = 0.564, P < 0.001). After propensity score adjustment, XELOX significantly increased overall 5-year survival compared to surgery only (58.2% vs 44.2%, P = 0.025) but not compared to FOLFOX6 therapy (48.5% vs 42.7%, P = 0.685). The incidence of grade 3/4 adverse reactions was similar between the XELOX and FOLFOX6 groups, and more patients suffered from hand-foot syndrome in the XELOX group (P = 0.018). CONCLUSION: Adjuvant XELOX therapy is associated with better survival in patients after D2 dissection, but does not result in a greater survival benefit compared with FOLFOX6 therapy.

Evaluation of skeletonization of the hepatoduodenal ligament for the lower third gastric cancer by propensity score analysis.

BACKGROUND/AIMS: Effects and indications of no. 12b and 12p nodes dissection for gastric cancer are not determined yet. Here we retrospectively evaluated the effect of no. 12b and 12p nodes dissection for treatment of lower third gastric cancer (LTGC). METHODOLOGY: Between 2001 and 2010, 110 LTGC patients with no. 12b and 12p nodes dissection (SHDL group) and 138 patients without no. 12b and 12p nodes dissection (non-SHDL group) were enrolled in this study. Clinicopathological features and prognostic data were compared between the two groups. RESULTS: The nodal metastatic rate was 8.2% of no. 12b and 10.9% of no. 12p. The 5-year survival rate was 62.9% in the SHDL group and 51.4% in the non-SHDL group (p = 0.16). Multivariate analysis with and without propensity score adjustment showed that SHDL was a significantly prognostic factor. The hazard ratio for death after D2 surgery plus SHDL was 0.457 (95% CI: 0.25 to 0.821; p = 0.0085) compared to D2 surgery alone. More patients in the non-SHDL group had only lymph node recurrence compared to the SHDL group (4.3% vs. 0%, p = 0.035). CONCLUSIONS: Skeletonization of the hepatoduodenal ligament is associated with superior outcomes for LTGC patients especially for those with involved local hepatoduodenal nodes.