Nevin Stage Research Articles

Novel immune scoring dynamic nomograms based on B7-H3, B7-H4, and HHLA2: Potential prediction in survival and immunotherapeutic efficacy for gallbladder cancer.

BackgroundGallbladder cancer (GBC) is a mortal malignancy with limited therapeutic strategies. We aimed to develop novel immune scoring systems focusing on B7-H3, B7-H4, and HHLA2. We further investigated their potential clinical effects in predicting survival and immunotherapeutic efficacy for GBC.MethodsThis was a retrospective cohort study in a single center that explored the expression characteristics of B7-H3, B7-H4, and HHLA2. The immune scoring nomograms for prognostic were developed via logistic regression analyses. Their performance was evaluated using the Harrell concordance index (C-index) and decision curves analysis (DCA), and validated with calibration curves.ResultsB7-H3, B7-H4, and HHLA2 manifested with a relatively high rate of co-expression patterns in GBC tissues. They were associated with worse clinicopathological stage, suppression of immune microenvironment, and unfavorable prognosis in postoperative survival. B7 stratification established based on B7-H3, B7-H4, and HHLA2 was an independent prognostic predictor (p<0.05 in both groups). Moreover, immune stratification was also successfully constructed based on B7 stratification and the density of CD8+ TILs (all p<0.001). The prediction models were developed based on B7-/or immune stratification combined with the TNM/or Nevin staging system. These novel models have excellent discrimination ability in predicting survival and immunotherapeutic efficacy for GBC patients by DCA and clinical impact plots. Finally, dynamic nomograms were developed for the most promising clinical prediction models (B7-TNM model and Immune-TNM model) to facilitate prediction.ConclusionsImmune scoring systems focusing on B7-H3, B7-H4, and HHLA2 may effectively stratify the prognosis of GBC. Prognostic nomograms based on novel immune scoring systems may potentially predict survival and immunotherapeutic efficacy in GBC. Further valid verification is necessary.

HHLA2 promotes tumor progression by long non‑coding RNA H19 in human gallbladder cancer.

Advanced gallbladder cancer (GBC) is one of the most malignant of all types of biliary tract cancers that is associated with poor prognosis and high mortality. Accumulating evidence suggest that the B7 family of proteins serve an essential role in various types of cancers, including GBC. However, the potential function and regulatory mechanism of human endogenous retrovirus-H long terminal repeat-associating protein 2 (HHLA2; also known as B7-H7 or B7H5) in GBC remain poorly understood. In the present study, immunohistochemistry was used to examine the expression pattern of HHLA2 in samples from 89 patients with GBC. The possible association between HHLA2 expression and the clinicopathological parameters, including prognosis, were then assessed. Using lentiviruses, overexpression of HHLA2 plasmid or short-hairpin RNA (shRNA) of HHLA2 were transfected into GBC-SD cells to overexpress or knock down HHLA2 expression, respectively. The effects of HHLA2 overexpression and knockdown on the epithelial-mesenchymal transition (EMT) process on GBC-SD cells were measured by the western blotting and immunofluorescence staining of collagen I, N-cadherin, E-cadherin, vimentin and α-SMA. By contrast, changes in cell proliferation were measured using EdU assay. Cell invasion and migration were assessed using Transwell and wound-healing assays, respectively. In addition, a xenograft mouse model was established to evaluate the tumorigenic ability of the GBC cell line in vivo after stable transfection with lentivirus for HHLA2 overexpression or shRNA for HHLA2 knockdown. The regulatory relationships among TGF-β1, long non-coding RNA (lncRNA) H19 (H19) and HHLA2 were then investigated. The mRNA expression of lncRNA H19 were assessed using reverse transcription-quantitative PCR, whereas the expression levels of HHLA2 were detected by western blotting and immunofluorescence staining. HHLA2 expression was found to gradually increase as the stages of the GBC samples become more advanced. In addition, the expression level of HHLA2 was calculated to be positively associated with the Nevin stage, American Joint Committee on Cancer stage, tumor invasion and regional lymph node metastasis but was negatively associated with the overall patient survival (OS). In vitro experiments demonstrated that overexpression of HHLA2 promoted GBC migration, invasion, proliferation and EMT, whereas in vivo experiments found a promoting role of HHLA2 overexpression on GBC tumor growth. After transfection with lentiviruses encoding the overexpression plasmid of lncRNA H19, GBC migration, invasion, proliferation and EMT were increased. By contrast, knocking down HHLA2 expression suppressed TGF-β1- or lncRNA H19 overexpression-induced GBC migration, invasion, proliferation and EMT. In addition, HHLA2 knockdown significantly reduced the sizes of the GBC tumors in vivo. These results suggest that HHLA2 overexpression can promote GBC progression. Conversely, ablation of HHLA2 expression inhibited both TGF-β1- and lncRNA H19-induced GBC progression, suggesting that HHLA2 is a potential therapeutic target for this disease.

Development and validation of a prognostic nomogram for gallbladder cancer patients after surgery.

BackgroundGallbladder cancer is associated with late diagnosis and poor prognosis. Current study aims to develop a prognostic nomogram for predicting survival of gallbladder cancer patients after surgery.MethodsTwo large cohorts were included in this analysis. One consisted of 1753 gallbladder cancer patients from the Surveillance, Epidemiology, and End Results (SEER) database, and the other consisted of 239 patients from Shanghai Renji hospital. Significant prognostic factors were identified and integrated to develop the nomogram. Then the model was subjected to bootstrap internal validation and external validation.ResultsUnivariate and multivariate analysis indicated that age, tumor histology, T-stage, N-stage and M-stage were significant prognostic factors, which were all included to build the nomogram. The model showed good discrimination, with a concordance index (C-index) of 0.724 (95% CI, 0.708–0.740), and good calibration. Application of the nomogram in the validation cohort still presented good discrimination (C-index, 0.715 [95% CI 0.672–0.758]) and good calibration. In the primary cohort, the C-index of the nomogram was 0.724, which was significantly higher than the Nevin staging system (C-index = 0.671; P < 0.001) and the 8th TNM staging system (C-index = 0.682; P < 0.001). In the validation cohort, the C-index of the nomogram was 0.715, which was also higher than the Nevin staging system (C-index = 0.692; P < 0.05) and the 8th TNM staging system (C-index = 0.688; P = 0.06).ConclusionsThe proposed nomogram resulted in more-accurate prognostic prediction for patients with gallbladder cancer after surgery.

A Novel Prognostic Nomogram for Gallbladder Cancer after Surgical Resection: A Single-Center Experience.

Background Gallbladder cancer (GBC), which accounts for more than 80% of biliary tract malignancies, has a poor prognosis with an overall 5-year survival less than 10%. The study aimed to identify risk factors and develop a predictive model for GBC following surgical resection. Methods 98 GBC patients who underwent surgical resection from Guangdong Provincial People's Hospital were enrolled in the study. Cox-regression analysis was performed to identify significant prognostic factors. A nomogram was constructed and Harrell's concordance index, calibration plot, and decision cure analysis were used to evaluate the discrimination and calibration of the nomogram. Results Liver resection, tumor size, perineural invasion, surgical margin, and liver invasion were identified as independent risk factors for overall survival (OS) in GBC patients who underwent surgical resection. Based on the selected risk factors, a novel nomogram was constructed. The C-index of the nomogram was 0.777, which was higher than the American Joint Committee on Cancer (AJCC) staging system (0.724) and Nevin staging system (0.659). Decision cure analysis revealed that the nomogram had a better net benefit and the calibration curves for the 1-, 3-, and 5-year survival probabilities were also well matched with the actual survival rates. Lastly, high-risk GBC were stratified based on the scores of the nomogram and we found high-risk GBC were associated with both worse OS and disease-free survival (DFS). Conclusion We developed a nomogram showing a better predictive capacity for patients' survival of resected GBC than the AJCC staging systems. The established model may help to stratify high-risk GBC and facilitate decision-making in the clinic.

Clinical significance of serum miR-187 and miR-143 in the diagnosis of gallbladder cancer

Objective To investigate the clinical significance of serum miR-187 and miR-143 in the development and diagnosis of gallbladder cancer. Methods 75 serum samples in patients with gallbladder cancer were selected as gallbladder cancer group. 75 serum samples in patients with gallbladder benign disease and 45 serum samples in healthy physical examinations at same period were selected as the benign gallbladder disease group and healthy control group. Quantitative RT-PCR was used to detect the serum miR-187 and miR-143 expression in each group, and the expression of those related with the clinicopathological factors, the proliferation and migration of gallbladder cancer cells, and the efficacy in diagnosis of gallbladder cancer was observed. Results The serum miR-187 expression in gallbladder cancer group was significantly higher than that in benign gallbladder disease and healthy control; the serum expression of that in benign gallbladder disease was significantly higher than that in healthy control; after surgery , the expression of that was significantly lower than that before treatment (all P 0.05), and were significantly correlated with Nevin stage, TNM stage, differentiation and lymphatic metastasis (all P<0.05). Furthermore, it was confirmed that miR-187 promoted the proliferation and migration of gallbladder cancer cells in vitro, while miR-143 inhibited the proliferation and migration. In the diagnosis of gallbladder, the diagnostic efficacy of miR-187 and miR-143 was significantly better than that of CA199 and CA242 (both P<0.05). Combined detection could further improve the efficacy in diagnosis of gallbladder cancer. Conclusions miR-187 and miR-143 are involved in the development of gallbladder cancer. Combined detection of serum miR-187 and miR-143 in gallbladder cancer has a high diagnostic efficiency in the diagnosis of gallbladder cancer. Key words: Gall bladder neoplasm; MiR-187; MiR-143; Diagnosis; Real-time -PCR

Clinical Analysis of 15 Cases of Gallbladder Neuroendocrine Carcinoma and Comparison with Gallbladder Adenocarcinoma Using a Propensity Score Matching.

PurposeThis study aimed to investigate the clinicopathological features and prognosis of gallbladder neuroendocrine carcinoma (GB-NEC).Patients and MethodsFifteen patients with GB-NEC and 171 patients with gallbladder adenocarcinoma (GB-ADC) treated in two tertiary medical centers between 2009 and 2015 were included. The clinicopathological features and prognostic risk factors of GB-NEC were analyzed retrospectively. A propensity score matching in a 1:2 ratio was used to compare the prognosis of GB-NEC and GB-ADC.ResultsFor patients with GB-NEC, the median age of patients was 58.4 years (range 26–75), with a M:F ratio of 7:8. Based on 2010 WHO classification, ten cases were pathologically confirmed as NECs and five cases as MANECs. For TNM staging, eleven patients were stage III or above; while for Nevin staging, seven patients were stage IV or above. The 1-, 2-, and 3-year overall survival (OS) of GB-NEC were 60.0%, 38.8% and 31.1%, respectively, and the median survival time was 20.4 months. Patients with lymph node metastasis had significantly shorter survival than those without (OS: 10.4 vs 26.0 months, p<0.05). Accordingly, patients of Nevin stage III had better OS than those of Nevin stage IV (p<0.05), but other potential risk factors including gender, age, clinical symptoms, TNM stage, histopathologic subtype and treatment showed no significance. After the propensity score matching, the baseline variables had no significant difference between 15 patients with GB-NEC and 30 patients with GB-ADC, survival analysis showed GB-NEC had worse prognosis (3-year overall survival rate: 31.1% vs 63.8%, p<0.01).ConclusionNevin staging helps classify patients of GB-NEC with different prognosis and the lymph node metastasis is a strong negative prognostic factor for OS. The propensity score analysis revealed even with the similar stage and treatment, GB-NEC still had worse OS than GB-ADC.

Epidemiologic analysis of gallbladder cancer

Background: Gallbladder cancer is characterized by poor prognosis and late diagnosis. Objective: The aim of our study is to describe the epidemiological variables of gallbladder cancer at our centre and to compare them with data from the worldwide literature. Material and methods: Patients undergoing cholecystectomy between January 1, 2006, and December 31, 2015. The following variables were analyzed: sex, age, male to female ratio, reason for cholecystectomy, urgent or scheduled. In case of gallbladder cancer, sex, age, histopathology, cell differentiation, gallbladder layer invasion, lymphovascular, cystic lymph node, adjacent tissues and liver invasion; and cancer staging according to Nevin staging system and the Union for International Cancer Control (UICC). Results: Ninety-two gallbladder neoplasms were found, representing 1.26% of all cholecystectomies. Mean age of patients with gallbladder cancer was 57.9 years (range, 22-88 years) of whom 69.5% (n=64) were women, with the same female to men ratio of 2:1 described for cholecystectomy. The histopathology of the 92 gallbladder cancers corresponded to adenocarcinoma, 93.4% (n= 86); squamous cell carcinoma, 4.4% (n= 4); and undifferentiated malignancy with elements of epithelial and mesenchymal origin, 2.2% (n= 2). There were two cases of adenocarcinoma with squamous metaplasia, two mucinous carcinomas, one papillary adenocarcinoma and one case of antral-type and intestinal metaplasia. Sixty tumors (65.4%) were moderately differentiated; 12 (13%) well differentiated; 11 (11.9%) poorly differentiated; and 4 (4.3%) were undifferentiated. In 5 cases (5.4%) the type of differentiation was not described. Conclusion: In gallbladder cancer diagnosed after a cholecystectomy, it is more frequent in the female sex, with adenocarcinoma being the most common histological type. Interestingly, the number of young patients with gallbladder cancer is noteworthy.

Kindlin-2 promotes gallbladder cancer metastasis and invasion by inducing epithelial-mesenchymal transition

Objective: To investigate the effects of Kindlin-2 on malignant phenotypes of human gallbladder cancer cells and discuss the mechanisms. Methods: The expression level of Kindlin-2 in 30 cases of gallbladder cancer tissues and adjacent non-tumoral tissues collected from the First Affiliated Hospital of Zhengzhou University between September 2012 and May 2013 was assessed by real-time PCR and immunohistochemistry.Lentivirus-mediated Kindlin-2 overexpression was used in gallbladder cancer cell lines GBC-SD and SGC-996.Transwell assay and adhesion assay were investigated to explore the functional role of Kindlin-2 on gallbladder cancer cells.Western Blot was used to test the protein change of epithelial-mesenchymal transition(EMT) characteristics. The t-test was used to analyzed results. Results: The RNA and protein levels of Kindlin-2 in gallbladder cancer tissues were higher than in the non-tumoral tissues (t=4.372, P=0.001; t=7.477, P=0.000). The expression level of Kindlin-2 in gallbladder cancer tissues was correlated with Nevin stage(χ(2)=5.932, P=0.035). Compared with control groups, the cell-matrix adhesion ability of GBC-SD and SGC-996 with Kindlin-2 overexpression was obviously promoted(1.66±0.03 vs. 1.07±0.22, t=2.710, P=0.041; 2.66±0.24 vs. 1.03±0.02, t=6.610, P=0.020). The number of GBC-SD and SGC-996 cells with Kindlin-2 overexpression passing through the Transwell chamber matrix increased significantly compared with the control groups(116.1±13.9 vs. 54.7±8.4, t=3.781, P=0.019; 136.3±7.5 vs. 64.3±6.4, t=7.302, P=0.002). The wound healing rate of GBC-SD with Kindlin-2 overexpression at 12-hour and 24-hour was higher than that of the group ((42.9±2.2)% vs. (29.7±1.7)%, t=4.690, P=0.009; (65.0±2.4)% vs.(40.4±2.0)%, t=7.945, P=0.001). The wound healing rate of SGC-996 with Kindlin-2 overexpression at 12-hour and 24-hour was also higher than that of the group ((32.9±1.3)% vs. (24.1±1.5)%, t=4.518, P=0.011; (51.3±1.1)% vs. (39.2±1.1)%, t=8.001, P=0.001). The characteristics of EMT were induced in gallbladder cancer cells with Kindlin-2 overexpression, including the up-regulation of N-cadherin, Vemintin and the down-regulation of E-cadherin. Conclusion: The expression of Kindlin-2 is up-regulated in gallbladder cancer tissues and Kindlin-2 promoted the malignant phenotypes of gallbladder cancer cells partially by epithelial-mesenchymal transition.

High ubiquitin-conjugating enzyme E2s expression is an independent factor of unfavorable prognosis in patients with gallbladder cancer

Objective To investigate the expression of ubiquitin-conjugating enzyme E2s (UBE2S) in gallbladder cancer (GBC) tissues and its clinical significance. Methods The expression of UBE2S was examined in 85 GBC, 28 pre-malignant lesion and 15 benign lesion tissues of the gallbladder by immunohistochemistry, and was verified by western blotting. The relationship between UBE2S expression and clinicopathological characteristics and prognostic factors of GBC patients was analyzed. Results Immunohistochemistry showed that UBE2S was highly expressed in GBC tissues compared to pre-malignant lesion tissues [stain index (SI): 4.082±3.413 vs. 2.607±2.025, P=0.021)] and benign lesion tissues (SI: 4.082±3.413 vs. 2.067±1.335, P=0.001), and western blotting showed that UBE2S protein was highly expressed in GBC tissues compared to benign lesion tissues (grey-scale value: 0.734±0.181 vs. 0.197±0.099, P=0.002). UBE2S was positively correlated with tumor differentiation degree (χ2=6.978, P=0.031) and liver metastasis (χ2=8.155, P=0.004). Cox model showed that UBE2S expression (P=0.024) as well as tumor differentiation degree (P=0.000), Nevin stage (P=0.040) and R0 resection (P=0.011) were prognostic factors of GBC patients. High UBE2S expression patients with GBC have a shorter survival time (P=0.011). Conclusion High UBE2S expression is an independent factor of unfavorable prognosis in GBC patients. Key words: Ubiquitin-conjugating enzyme E2s; Gallbladder neoplasm; Prognosis

Relationships between peripheral circulating tumor cells in patients with gallbladder carcinoma detected by a novel nano microfluidic chip with clinicopathology and prognosis

Objective To establish a novel method using a nano microfluidic chip to detect circulating tumor cells (CTCs) in peripheral blood in gallbladder carcinoma, and to study the relationship between CTCs with clinicopathology and prognosis in these patients. Methods The peripheral blood samples of 51 patients with gallbladder carcinoma were collected from June 2014 to January 2017. The CTCs from peripheral blood samples were detected by a novel nano microfluidic chip. This study aimed to study the correlation between CTCs with the clinical and pathological features. The significance of CTCs on prognosis in patients with gallbladder carcinoma was also analyzed. Results The positive rate of CTCs in the peripheral blood of gallbladder carcinoma patients was 43.1% (22/51). There were significant correlations between CTCs with liver metastasis (P 0.05). Conclusions Peripheral blood CTCs in patients with gallbladder carcinoma were efficaciously detected by a novel nano microfluidic chip. Peripheral blood CTCs was closely related to the Nevin staging and liver metastasis. CTCs could serve as a potential prognostic indicator in patients with gallbladder carcinoma. Key words: Gallbladder carcinoma; Microfluidic chip, nano; Circulating tumor cells (CTCs); Prognosis

Ring finger protein 125, as a potential highly aggressive and unfavorable prognostic biomarker, promotes the invasion and metastasis of human gallbladder cancers via activating the TGF- β1-SMAD3-ID1 signaling pathway.

Human gallbladder cancer (GBC) is a lethal aggressive malignant neoplasm. Identification of potential molecular biomarkers and development of targeted therapeutics for GBC patients is very necessary. In this study, we firstly investigated the correlation between ring finger protein 125 (RNF125) expression and the metastasis and prognosis of GBC, and the underlying molecular mechanism. RNF125 expression in a cohort of GBC tissues was examined; its correlation with clinicopathological and prognostic factors of GBC patients was analyzed. Moreover, the metastasis-related difference expressed genes in highly and lowly aggressive GBC cell lines were identified; and the influence of RNF125 knockdown on the metastatic phenotypes and characteristic EMT markers in highly aggressive GBC NOZ cells was detected. Furthermore, the underlying molecular mechanism of RNF125 effect was explored. The results showed that RNF125 was highly expressed in GBC tissues and related with aggressive characteristics such as Nevin stage (P = 0.041) etc. and unfavorable prognosis of GBC patients (P = 0.023, log-rank test). And, RNF125 was proved to a positive metastasis-related gene in vitro. RNF125 knockdown inhibited the invasion and migration, enhanced the adhesion, upregulated E-cadherin and β-catenin expression, and downregulated vimentin and N-cadherin expression (all P < 0.001) of NOZ cells in vitro. RNF125 promoting effect on GBC tumor progression was identified to relate with the activation of TGF-β1-SMAD3-ID1 signaling pathway. These findings firstly confirm that high RNF125 expression is related with aggressive characteristics and unfavorable prognosis of GBC patients; RNF125 promotes the invasion and metastasis of human GBCs via activating the TGF-β1-SMAD3-ID1 signaling pathway.

Lysine-specific demethylase 1 promotes tumorigenesis and predicts prognosis in gallbladder cancer.

Gallbladder Cancer (GBC), characterized by invasive growth and infiltrative dissemination, is difficult to diagnose and has poor prognosis. Emerging evidence demonstrates that Lysine-Specific Demethylase 1 (LSD1) has important roles in carcinogenesis, proliferation and metastasis. We studied the roles and molecular mechanisms of LSD1 in GBC. We examined LSD1 expression in 109 paired samples of GBC and normal gallbladder tissues. We found GBC tissues had upregulated LSD1 compared with normal gallbladder tissues (P = 0.003), and its high expression was associated with tumor-node-metastasis stage (P < 0.0001), Nevin's stage (P = 0.0093) and distant metastases (P = 0.0070). We found positive correlations between LSD1 expression and other proteins: epithelial-mesenchymal transition markers, C-myc and cyclin-related proteins. Inhibiting LSD1 expression in vitro impaired the proliferation and invasiveness of GBC cells and also downregulated c-myc expression and consequently inhibited GBC cell proliferation. LSD1 overexpression promotes GBC development and may be a predictor for a worsened prognosis. LSD1 may be a novel therapeutic target and prognostic tool for gallbladder cancer.

Prognostic significance of preoperative neutrophil/lymphocyte ratio and platelet/lymphocyte ratio in patients with gallbladder carcinoma.

Neutrophil/lymphocyte ratio (NLR) and platelet/lymphocyte ratio (PLR) were immune response-related indicators. Preoperative NLR and PLR had been considered to be related to the prognosis of various cancers. The objective of this study was to evaluate the prognostic significance of NLR and PLR in patients with gallbladder carcinoma (GBC). From 2001 to 2013, 145 patients with GBC were recruited in this retrospective study. Cutoff values of NLR and PLR were determined by receiver operating characteristic curves (ROC). The correlation of clinical data, including tumor differentiation, nevin stage, TNM stage, operation margin, operation mode, NLR, PLR, hemoglobin, C reactive protein (CRP), carcinoembryonic antigen (CEA), and carbohydrate antigen 199 (CA199) with median survival period of patients was analyzed by univariate survival analysis. The multivariate prognosis analysis was performed to select the independent prognostic factors. The cutoff values of NLR and PLR were 1.94 and 113.34, respectively. Compared with low NLR and low PLR group, the 5-year survival rates in high NLR and high PLR group were reduced (P < 0.05). The degree of tumor differentiation, nevin stage, TNM stage, operation mode, NLR, PLR, CA199, total bilirubin, CRP and CEA were associated with the median survival period of patients (P < 0.01). The multivariate prognosis analysis showed that NLR, nevin stage, operation mode and hemoglobin were independent prognostic factors (P < 0.05). Preoperative NLR and PLR were closely related to prognosis of patients with GBC and might be useful for the evaluation of prognosis of patients with GBC.

Prognostic significance of preoperative platelet count in patients with gallbladder cancer.

To investigate the prognostic value of preoperative platelet count (PLT) in patients with primary gallbladder cancer (GBC). The clinical data of 223 GBC patients after surgery was retrospectively reviewed. A receiver operating characteristic (ROC) curve was plotted to verify the optimum cutoff point for PLT. Univariate and multivariate survival analyses were performed to identify the factors associated with the prognosis. The ROC curve showed that the optimum cutoff point for PLT was 178 × 10(9)/L, and the entire cohort was stratified into group A with PLT > 178 × 10(9)/L and group B with PLT ≤ 178 × 10(9)/L. Group A had a better survival than group B (P < 0.001). There was an obvious difference between the two groups in terms of the differentiation degree, advanced tumor stage, lymph node metastasis (P < 0.001) and pathological type (P < 0.05). The univariate analysis demonstrated that tumor location, differentiation degree, TNM stage, Nevin stage, lymph node metastasis and PLT were associated with overall survival (P < 0.001). In the multivariate analysis, PLT (P = 0.032), lymph node metastasis (P = 0.007), tumor location (P < 0.001) and TNM stage (P = 0.005) were independent prognostic factors. PLT is closely correlated with GBC prognosis and could be used to identify the population with a poorer prognosis after surgery.

Prognostic implications of SLIT and ROBO1 expression in gallbladder cancer.

SLIT, a secretory glycoprotein, and its receptor roundabout (ROBO) are expressed in several types of cancer and have been implicated in tumor angiogenesis. The purpose of this study was to determine the prognostic implications of SLIT and ROBO1 expression and their association with clinicopathologic characteristics in gallbladder cancer. A retrospective analysis of 109 consecutive patients who underwent primary gallbladder cancer resection was conducted. Univariate and multivariate models were used to analyze the effect of clinicopathologic factors on survival. Expression of SLIT and ROBO1 was evaluated by immunohistochemistry, and their association with clinicopathologic characteristics was analyzed using mean testing. Multivariate linear regression analysis was also applied to investigate the independent predictors of ROBO1 expression. Seventy-five patients were included in the post-resection survival analysis, with 1-year and 3-year overall survival rates of 60 and 40 %, respectively. Univariate analysis revealed that pN classification, pT classification, pM classification, liver involvement, perineural invasion, TNM staging, Nevin staging, and microscopic resection margins affect prognosis. Multivariate analysis confirmed that pN classification, liver involvement, and perineural invasion are independent prognostic factors. In the mean tests of 109 cases, the mean difference of SLIT immunoreactivity was significant according to the presence of gallstones (P = 0.003) and liver involvement (P = 0.005), while the mean difference of ROBO1 immunoreactivity was significant according to liver involvement (P < 0.001), TNM staging (P < 0.001), and Nevin staging (P < 0.001). Multivariate analysis of ROBO1 immunoreactivity showed that SLIT immunoreactivity and TNM stage (adjusted R (2) = 0.203) or SLIT immunoreactivity and Nevin stage (adjusted R (2) = 0.195) were independent predictors of ROBO1 expression. pN classification, liver involvement, perineural invasion, and pathologic stage are significant prognostic factors for gallbladder cancer survival. SLIT expression is associated with cholelithiasis and liver involvement, and ROBO1 expression is associated with liver involvement and pathologic stage. In addition, SLIT expression and pathologic stage predict ROBO1 expression independently.

Expression of extracellular matrix metalloproteinase inducer CD147 and matrix metalloproteinase-9 in gallbladder carcinoma and precancerous lesions

Objective To investigate the expression of extracellular matrix metalioproteinase inducer CD147 and matrix metalloproteinase-9 (MMP-9) in the gallbladder carcinoma and precancerous lesions and their correlation with occurrence,progress,and prognosis of gallbladder carcinoma.Methods Streptavid-in-peroxidase (SP) immunohistochemistry was used to detect the expression of CD147 and MMP-9 in surgically resected specimens of 42 gallbladder carcinomas,18 gallbladder epithelial atypical dysplasia Ⅱ-Ⅲ tissue samples,13 gallbladder epithelial atypical dysplasia Ⅰ tissue samples,15 gallbladder adenomas and 15 chronic cholecystitis tissue samples.Results The positive rate of CD147 (83.33％)and MMP-9 (88.10％) proteins was higher in gallbladder carcinomas than in gallbladder epithelial atypical dysplasia Ⅰ tissues (38.46％,30.77％),gallbladder adenomas (20.00％,26.67％) and chronic cholecystitis tissues (6.67％,20.00％) (P ＜0.01),but no significant difference was found in comparison to gallbladder epithelial atypical dysplasia Ⅱ-Ⅲ tissues (61.11％,66.67％,P ＞0.05).The expression of CD147 and MMP-9 was positively correlated with the tumor differentiation,Nevin staging and lymph node metastasis (P ＜0.05).The CD147 expression was positively correlated with MMP-9 expression in gallbladder carcinoma (rs =0.427,P ＜0.01).Patients negative for the expression of CD147 and MMP-9 had longer survival time than those with positive expression of CD147 and MMP-9.Conclusion CD147 and MMP-9 play important roles in carcinogenesis of gallbladder mucosa,and invasion,metastasis and prognosis of gallbladder carcinoma. Key words: Gallbladder carcinoma; Atypical dysplasia; CD147; Matrix metalloproteinase

The expression of Nek7, FoxM1, and Plk1 in gallbladder cancer and their relationships to clinicopathologic features and survival

Gallbladder carcinoma (GC) is generally considered as a relatively rare malignancy with poor prognosis. In order to guide clinicians in selecting suitable treatment for GC patients, reliable markers predictive of poor clinical outcome are desirable. This study analyzed the expression of Polo-like kinase 1 (Plk1), Nima related kinases 7 (Nek7) and Forkhead box M1 (FoxM1) in GC tissues and their relationship to clinicopathologic features and survival. We immunohistochemically investigated the 76 specimens of gallbladder carcinoma, pericarcinoma and normal tissues using Nek7, FoxM1 and Plk1 antibodies and analyzed the overall survival time of these 76 patients. There were significant correlations between the high level expression of Nek7, FoxM1 and Plk1 and the tumor differentiation, Nevin staging and metastasis. The high level expression of Nek7, FoxM1 and Plk1 was significantly associated with shorter overall survival time in univariate analysis (log-rank test), also identified as an independent prognostic factor in multivariate analysis. Nek7, FoxM1 and Plk1 were significantly associated with certain clinicopathologic indices in GC. Evaluation of Nek7, FoxM1 and Plk1 expression may be an important factor in identifying a group of poor GC prognosis.

Expression of cyclooxygenase-2 and vascular endothelial growth factor C and its clinicopathological significance in adenoma and adenocarcinoma of gallbladder

Objective To study the expression levels of cyclooxygenase-2 (COX-2) and vascular endothelial growth factor C (VEGF-C) and their clinicopathological significance in benign and malignant lesions of gallbladder.Methods The expression levels of COX-2 and VEGF-C were detected by using immunohistochemistry in 44 cases of gallbladder cancer and 10 cases of gallbladder adenoma.Results The expression rate of COX-2 in gallbladder adenocarcinoma and adenoma was 72.7％ ％ and 40.0％ respectively.The expression rate of COX-2 in gallbladder adenocarcinoma was higher than that in gallbladder adenoma (P ＜ 0.05).The expression rate of VEGF-C in gallbladder adenocarcinoma was higher than that in gallbladder adenoma (63.6％ vs.20.0％,P ＜ 0.05).There was a significant correlation between the expression of COX-2,VEGF-C and Nevin stages and lymph node metastasis of gallbladder carcinoma.There was a significant relationship between COX-2 and VEGF-C.Conclusion The expression of COX-2 and VEGF-C might be closely related to the carcinogenesis and biological behaviors of gallbladder adenocarcino Key words: Gallbladder neoplasms; Cyclooxygenase-2; Vascular endoehelial growth factor C

Gallbladder Carcinoma: Analysis of Prognostic Factors in 132 Cases

To evaluate the prognostic factors of gallbladder carcinoma. Presentation, operative data, complications, and survival outcome were examined for 132 gallbladder carcinoma patients who underwent gallbladder surgery in our unit during 2002-2007, and follow-up results were obtained from every patient for univariate and multivariate survival analysis. The univariate analysis showed that gallbladder lesion history, tumor cell differentiation, Nevin staging, preoperative lymph node metastasis and the surgical approach significantly correlated with the prognosis of the patients (p <0.05). The results of the multivariate analysis (Cox regression) showed that gallbladder lesion history, Nevin staging and the surgical approach were independent predicators with relative risks of 6.9, 4.4, 2.8, respectively (p=0.002, 0.003, 0.008). Gallbladder lesion history, Nevin staging and the surgical approach are independent prognostic factors for gallbladder carcinoma, a rapidly fatal disease. Therefore, early diagnosis, anti-infective therapy and radical surgery are greatly needed to improve the prognosis of gallbladder carcinoma.

Expression differences of Cathepsin B and Cathepsin D in primary gallbladder cancer, gallbladder adenomatous polyps and chronic cholecystitis

Objective To investigate the expression of cathepsin B (CatB) and cathepsin D (CatD) in primary gallbladder carcinoma,adenomatoid polyp without malignant transformation,and chronic cholecystitis tissues and to assess their mutual correlation ships as well as the relationships between their expressions and clinical pathological features.Methods Detected the expressions of CatB and CatD in gallbladder tissues including 32 specimens of primary gallbladder carcinoma,10 specimens of adenomatoid polyp without malignant transformation and 10 specimens of chronic cholecystitis by immunohistochemical method.The expressions were assessed by semi-quantitative analysis.Results The positive rate of CatB,CatD and CCL20 in primary gallbladder carcinoma was 93.75％ (30/32) and 90.63％ (29/32) respectively,being significantly higher than those in adenomatoid polyp without malignant transformation (60％and 60％,P ＜0.05) and chronic cholecystitis groups (60％ and 50％ P ＜0.05).The expression levels of CatB and CatD were not significantly related to the clinical features including age and gender,or pathological features including grades of cell differentiation,pathological types and Nevin stages ( P ＞ 0.05 ).CatB and CatD had significantly positive correlations ( r =0.434,P ＜ 0.05 ).Conclusion ( 1 ) CatB and CatD CCL20 might serve as the histological marker of primary gallbladder carcinoma.(2) The expression levels of CatB and CatD in primary gallbladder carcinoma are not related to the clinical or pathological features.(3) There is positive correlation between the expression levels of CatB and CatD in primary gallbladder carcinoma. Key words: Primary gallbladder carcinoma; Cathepsin B; Cathepsin D; Immunohistochemistry