Pathogenesis Of Gallbladder Cancer Research Articles

Targeting gallbladder cancer: a pathway based perspective.

Gallbladder cancer (GBC) has a poor prognosis with a 5-year survival rate suggesting the need for more effective treatment strategies. Studying the cross-talk of several pathways involved in crucial cellular and biological processes such as cell growth, proliferation, migration and apoptosis would prove beneficial in identifying key players of GBC progression and targeting them. This review highlights several pathways known to be dysregulated in GBC onset and progression and describes known and potential targets. Within these pathways, there are proteins involved in the signalling cascade, which may be targeted as potential biomarkers and drug targets. Furthermore, the cross-talk of these pathways is investigated in the context of GBC and the implications thereof. A better understanding of the pathways involved in GBC pathogenesis will aid clinicians in the prognosis, diagnosis and treatment of patients. There are significant clinical implications of GBC pathway-based studies as they permit the understanding of onset and progression of the disease.

Overexpression of EIF5A2 is associated with poor survival and aggressive tumor biology in gallbladder cancer.

Gallbladder cancer (GBC) is a malignant tumor of the biliary tract. The main problem affecting the treatment of gallbladder cancer is late diagnosis and poor prognosis. EIF5A2 is one of two isoforms of the EIF5A family and is reported to be a new oncogenic protein in many human cancers. In this study, our results showed for the first time that EIF5A2 was overexpressed in GBC samples compared with non-tumor tissue. Overexpression of EIF5A2 was associated with lymph node metastasis, tumor differentiation, UICC (Union for International Cancer Control) staging, histological type, metastasis, and tumor size. Overexpression of EIF5A2 in gallbladder carcinoma tissues is also associated with poor prognosis in patients. The interference of EIF5A2 significantly inhibited the proliferation, cell cycle, migration and colony formation of GBC-SD cells in vitro. Our results suggest that EIF5A2 is a target oncogene and may be an important prognostic biomarker in the pathogenesis of gallbladder cancer.

Identification of messenger and long noncoding RNAs associated with gallbladder cancer via gene expression profile analysis.

The present study aimed to investigate the long noncoding RNAs (lncRNAs) and messenger RNAs (mRNAs) involved in the progression of gallbladder cancer and explore the potential physiopathologic mechanisms of gallbladder cancer in terms of competing endogenous RNAs (ceRNAs). The original lncRNA and mRNA expression profile data (nine gallbladder cancer tissues samples and nine normal gallbladder samples) in GSE76633 was downloaded from the Gene Expression Omnibus database. Differentially expressed mRNAs and lncRNAs between gallbladder cancer tissue and normal control were selected and the pathways in which they are involved were analyzed using bioinformatics analyses. MicroRNAs (miRNAs) were also predicted based on the differentially expressed mRNAs. Finally, the co-expression relation between lncRNA and mRNA was analyzed and the ceRNA network was constructed by combining the lncRNA-miRNA, miRNA-mRNA, and lncRNA-mRNA pairs. Overall, 373 significantly differentially expressed mRNAs and 47 lncRNAs were identified between cancer and normal tissue samples. The upregulated genes were significantly enriched in the extracellular matrix (ECM)-receptor interaction pathway, while the downregulated genes were involved in the complement and coagulation cascades. Altogether, 128 co-expression relations between lncRNA and mRNA were obtained. In addition, 196 miRNA-mRNA regulatory relations and 145 miRNA-lncRNA relation pairs were predicted. Finally, the lncRNA-miRNA-gene ceRNA network was constructed by combining the three types of relation pairs, such as XLOC_011309-miR-548c-3p-SPOCK1 and XLOC_012588-miR-765-CEACAM6. mRNAs and lncRNAs may be involved in gallbladder cancer progression via ECM-receptor interaction pathways and the complement and coagulation cascades. Moreover, ceRNAs such as XLOC_011309-miR-548c-3p-SPOCK1 and XLOC_012588-miR-765-CEACAM6 can also be implicated in the pathogenesis of gallbladder cancer.

Epidemiology of gallbladder cancer in India.

India is a high incidence area for gallbladder cancer (GBC) and contributes to about 10% of the global GBC burden. Within India, the incidence is high in North, North-East, Central and Eastern India, and less common in South and West India. The incidence has been on a steady rise in both genders. The presentation is often with advanced disease and carries dismal prognosis. GBC in India usually affects younger patients in the 5th and 6th decade in contrast to the west. Gallstones are present in 80% of the Indian patients with GBC and its presence increases the vulnerability of the GB to mucosal injury. The incidence of GBC is out of proportion to the prevalence of gallstones in the country. Additional co-factors such as older age, lower socio-economic status, chronic Salmonella typhi (S. typhi) infection, Helicobacter pylori (H. pylori) infection, exposure to pollutants, heavy metals, chemicals, adulterated mustard oil and smoking in patients with gallstones have been identified which promote carcinogenesis. These risk factors act in tandem in an additive manner resulting in higher incidence of GBC as well as hasten the development of GBC. Environmental risk factors such as soil and water contamination by industrial wastes, agricultural effluents and human sewage have been identified as putative risk factors. Combination of a toxic environment, vulnerable GB and a susceptible host play a key role in the pathogenesis of GBC in the country. Large multicentric comprehensive studies are required in India to assess the attributable risk of each of the identified putative risk factors. This will help in formulating cost effective national strategies in preventing GBC related mortality in the country. Meanwhile a high index of suspicion to pick up incidental GBC, and improved access to healthcare facilities to manage GS appropriately will help in reducing GBC related mortality.

Characteristics of a novel cell line ZJU-0430 established from human gallbladder carcinoma

BackgroundGallbladder cancer is the most common malignant neoplasm of the biliary tract, responsible for 80–95% of cases. Appropriate models are required for investigating the molecular pathogenesis of gallbladder cancer.MethodsIn this study, we aimed to establish a gallbladder cancer cell line from primary tumour. Single cell RNA sequencing, Light and electron microscopy, DNA content analysis, cytogenetic analysis, short tandem repeat (STR) DNA fingerprint analysis, immunophenotypic characterization, and xeno-transplantation were utilized to characterize the novel ZJU-0430 cell line in vitro and in vivo.ResultsThe cell line showed multiple cell shapes and characteristic epithelial morphologies under the microscope, but no too much heterogeneity by scRNA-Seq, with a population doubling time (PDT) of 19.81 h, which was shorter than that for GBC-SD cells. An immunophenotypic analysis revealed that ZJU-0430 cells were positive for CD24, CD44, CD29 and CD133 expression, and partially positive for CD184, and CD326 expression, and negative for CD34, CD90, CD117, and CD338 expression, similar to the primary cancer cells. A pathological analysis confirmed the origination of cell line from gallbladder tumour. ZJU-0430 cells had higher migration, invasion and proliferation properties than GBC-SD cells in vitro, and showed in vivo tumorigenicity in nude mouse xenograft settings.ConclusionsThe results confirm the potential utility of ZJU-0430 cell line as a representative model of gallbladder cancer and suggest that it could be used in the in vitro and in vivo studies of gallbladder cancer pathogenesis and to develop new therapeutics.

Pathogenesis of gallbladder cancer and current status of laparoscopic cholecystic carcinoma surgery

Gallbladder carcinoma is a kind of malignant tumor induced by many factors with high mortality. The pathogenesis of gallbladder cancer is still unclear. Surgical resection is the first choice of treatment. With the development of minimally invasive technology, laparoscopy is more and more widely used in the treatment of gallbladder carcinoma with its unique advantages. However, the indication of laparoscopic surgery in patients with gallbladder cancer hasn′t been standardized around the world. By reviewing the relevant literature in recent years, the causes of gallbladder carcinoma and the problems related disputes in laparoscopic surgery are summarized and discussed in order to provide guidance and basis for the future laparoscopic cholecystic carcinoma surgery. Key words: Gallbladder neoplasms; Laparoscopes; Surgical procedures, operative

Advances in comprehensive diagnosis and treatment of gallbladder cancer

Gallbladder cancer is a highly malignant tumor of the digestive system, and its incidence in China has been increasing year by year. Due to the lack of specific symptoms in the early stage, early diagnosis is difficult for gallbladder cancer and it is often at advanced stage once discovered. Surgery is still the only method to cure gallbladder cancer. Therefore, early diagnosis is particularly important for radical resection and improving overall prognosis of gallbladder cancer patients. However, there is still no specific molecular diagnostic target for gallbladder cancer. The continuous improvement of research and surgical skills have made the operational procedure of gallbladder cancer more standardized. With the increasing understanding on the epidemiology and pathogenesis of gallbladder cancer and the improvement of imaging, radiotherapy and chemotherapy technology, the comprehensive diagnosis and treatment of gallbladder cancer has also developed rapidly in recent years. However, high-quality clinical research is still needed. Comprehensive treatment model including radical surgery, adjuvant radiotherapy and chemotherapy, targeted therapy and immunotherapy will be the trend of gallbladder cancer treatment in the future. This article summarizes the advances in comprehensive diagnosis and treatment of gallbladder cancer by reviewing literatures. Key words: Gallbladder neoplasms; Early diagnosis; Neoadjuvant therapy; Comprehensive diagnosis and treatment

The emerging roles of long non-coding RNA in gallbladder cancer tumorigenesis.

Accumulating evidence suggests that long non-coding RNAs (lncRNAs) have important regulatory functions in gallbladder cancer (GBC) tumorigenesis and can serve as potential novel markers and/or targets for GBC. In this review, we critically discuss the emerging alteration of lncRNAs in GBC, the lncRNAs induced epigenetic regulation, the interaction of lncRNAs with microRNAs and lncRNAs effects on tumor-related signaling pathways. Additionally, contributions of lncRNAs in epithelial-mesenchymal transition process and energy metabolism reprogramming in GBC are also addressed. This may pave new ways towards the determination of GBC pathogenesis and lead to the development of new preventive and therapeutic strategies for GBC.

C‑Jun suppresses the expression of WNT inhibitory factor 1 through transcriptional regulation and interaction with DNA methyltransferase 1 in gallbladder cancer.

WNT inhibitory factor 1 (WIF-1) is involved in the tumorigenicity and progression of several types of tumor, which has been attributed to aberrant hypermethylation of its promoter. However, the role of WIF-1 in the pathogenesis of gallbladder cancer (GBC) remains to be fully elucidated, and the data available are insufficient to identify the upstream molecular mechanisms involved. In the present study, the methylation status of the WIF-1 promoter was investigated using methylation-specific polymerase chain reaction (PCR) and bisulfate sequencing PCR in GBC cells. Immunohistochemistry, reverse transcription-quantitative PCR and western blotting were used to analyze the expression of WIF-1 and c-Jun. In addition, a co-immunoprecipitation assay was designed to determine the DNA methyltransferase that was implicated in WIF-1 methylation. The results revealed that the expression of WIF-1 was low in GBC, and that this was caused by aberrant DNA hypermethylation. However, there were no significant correlations between the expression of WIF-1 and certain key clinicopathological characteristics of GCB. Subsequently, a negative correlation was found between the protein expression of c-Jun and WIF-1 in 50 GBC specimens using immunohistochemistry. The demethylation and re-expression of WIF-1 was observed when the expression of c-Jun was silenced. Finally, it was found that the knockdown of c-Jun downregulated the expression of DNA methyltransferase 1 (DNMT1) and that c-Jun interacted with DNMT1. Taken together, the present study suggested that c-Jun suppressed the expression of WIF-1 through transcriptional regulation and interaction with DNMT1 in GBC. These findings provide an alternative pathogenesis of GBC, which may be promising as a novel reference for early diagnosis or future treatment.

Regional differences in gallbladder cancer pathogenesis: Insights from a comparison of tumor mutations

Regional differences in gallbladder cancer pathogenesis: Insights from a comparison of tumor mutations

Histologic and Molecular Pathogenesis of Gallbladder Cancer

담낭암의 ëŒ€í‘œì ì¸ 조직형은 담낭 ì ë§‰ì˜ 상피세포에서 기원하는 ì„ ì•”(adenocarcinoma)이며, í˜•íƒœì  ìœ ì‚¬ì„±ì— 따라 담도성(biliary), 장관성(intestinal), 위의 소와세포성(foveolar), 혹은 편평상피성(adenosquamous) 등으로 ë¶„ë¥˜í• ìˆ˜ 있다. ì„ ì•”ì˜ ì „ì•”ì„± 병변은 3가지가 있으며 1) ì„ ì¢ (adenoma), 2) ë‹´ë„ìƒí”¼ë‚´ì¢ ì–‘(BilIN), 3) 담낭내 ìœ ë‘ìƒ ì¢ ì–‘(intracystic papillary neoplasm)으로 êµ¬ë¶„í• ìˆ˜ 있다. 이런 ì „ì•”ì„± 병변은 ê³µí†µì ìœ¼ë¡œ ì„¸í¬í•™ì  이형성을 갖는 ì ë§‰ 상피 세포의 증식이라는 특징을 ê°–ê³ ìžˆìœ¼ë‚˜, ì¢ ì–‘ 세포가 ì ë§‰ 아래 간질을 침습하지 않은 상태의 병변이다. 내강 내로 돌출된 폴립 모양의 병변이나 ì ë§‰ì´ 과립상으로 비후된 경우, ì„ ì¢ í˜¹ì€ ë‹´ë‚­ë‚´ìœ ë‘ìƒ ì¢ ì–‘ì˜ 경우가 많으며, 육안상 확인되지 않으나 í˜„ë¯¸ê²½ì ìœ¼ë¡œ 상피 세포의 이형성이 ê´€ì°°ë ê²½ìš° ë‹´ë„ìƒí”¼ë‚´ì¢ ì–‘ì— 해당한다. 이런 ì „ì•”ì„± 병변은 이형성의 ì •ë„ì— 따라 ì €ë“±ê¸‰(low), 중등급(moderate), ê³ ë“±ê¸‰ 이형성(high grade dysplasia)의 3등급으로 í‰ê°€í•˜ê³ ìžˆìœ¼ë©°, ê³ ë“±ê¸‰ì˜ 경우 상피내암(carcinoma in situ)ê³¼ 동일한 병변으로 ì·¨ê¸‰í•˜ê³ ìžˆë‹¤. ì„ ì¢ ê³¼ 담낭내 ìœ ë‘ìƒ ì¢ ì–‘ì€ í˜•íƒœí•™ì  특징이 중복되는 부분이 있어, 진단의 재현성을 높이기 위한 í˜•íƒœí•™ì  기준 ë§ˆë ¨ì´ 필요한 병변이다. 담낭 ì„ ì•”ì—ì„œ ë³´ê³ ëœ 가장 ëŒ€í‘œì ì¸ ë¶„ìžìœ ì „í•™ì  변이는 KRAS, TP53ê³¼ CDKN2A가 ì•Œë ¤ì ¸ ìžˆê³ ERBB2의 증폭도 ì•Œë ¤ì ¸ 있으나, 담도암에 íŠ¹ì´ì ì´ë©´ì„œ 변이율이 높은 ìœ ì „ìžëŠ” 많지 않다. ì „ì•”ì„± 병변에서 ì„ ì•”ìœ¼ë¡œ 이어지는 암발생 ê³¼ì •ì—ì„œ ë‹¨ê³„ì ìœ¼ë¡œ 발견되는 ë¶„ìžìœ ì „ì  변이 또한 ì „ì•”ì„± 병변의 ì¢ ë¥˜ë‚˜ 만성 담낭염, 췌담도 기형 등의 위험인자 ìœ ë¬´ì— 따라 ë³´ê³ ìœ¨ì´ 서로 달라, 한 가지 ê¸°ì „ìœ¼ë¡œ ì„¤ëª í•˜ê¸°ëŠ” ì–´ë µë‹¤. 만성 담낭염을 ì„ í–‰ì¸ìžë¡œ 갖는 경우 ì ë§‰ 상피의 증식 혹은 화생의 초기 단계에서 COX-2 과발현이나 TP53 변이, ë§ˆì´í† ì½˜ë“œë¦¬ì•„ DNA 손상이 ë°œìƒí•˜ê³ , 초기 이형성 단계에서 3p, 8p 염색체의 ì´í˜•ì ‘í•©ì„± 소실(loss of heterozygosity, LOH) 및 HER2 증폭이 ë³´ê³ ë˜ê³ ìžˆê³ , ìƒí”¼ë‚´ì¢ ì–‘ì—ì„œëŠ” 9p, 18q, 22q, 17p 염색체의 LOH 및 CDK2A 변이가 관찰되는 것으로 ì•Œë ¤ì ¸ 있다. Keywords: 담낭암, ì„ ì•”, ì „ì•”ì„± 병변, 변이, ë°œì•”ê³¼ì •

Dysregulated Expression and Subcellular Localization of Base Excision Repair (BER) Pathway Enzymes in Gallbladder Cancer.

Base excision repair (BER) pathway is one of the repair systems that has an impact on radiotherapy and chemotherapy for cancer patients. The molecular pathogenesis of gallbladder cancer is not known extensively. In the present study we investigated whether the expression of AP endonuclease 1 (APE1) and DNA polymerase β (DNA pol β), key enzymes of BER pathway has any clinical significance with gallbladder carcinogenesis. 41 gallbladder cancer, 27 chronic cholecystitis, and 3 normal gallbladder specimens were analyzed for the expression of APE1 and DNA polymerase β by western blotting, and subcellular localization studied by immunohistochemistry. The enzymatic activity of APE1 was also studied. The correlations with expression of the above proteins with clinical-pathological characteristics of gallbladder cancer patients were analyzed. The integrated density value ratio (relative expression) of total APE1 (37 kDa + 35 kDa variant) analyzed in the three groups of tissues, was 0.76±0.03 in normal gallbladder, 0.91±0.08 in chronic cholecystitis, and 1.12±0.05 in gallbladder cancer. APE1 was found to be up-regulated in 80% of gallbladder carcinoma samples (P = 0.01). A positive trend of APE1 expression with tumor stage and lymph node positivity was observed. The enzymatic activity of APE1 was found higher in gallbladder cancer samples in comparison with chronic cholecystitis. The integrated density value ratio of DNA polymerase β for normal gallbladder, chronic cholecystitis and gallbladder cancer tissue samples were 0.46±0.03, 0.7±0.06 and 1.33±0.1, respectively. DNA polymerase β was found to be upregulated in almost all gallbladder carcinoma samples (P =0.0001), and its expression was negatively correlated with age (P=0.02). DNA polymerase β expression showed a positive trend with tumor stage and nuclear differentiation of gallbladder cancer. It may be concluded that alteration of these BER pathway proteins may be the causal factors for carcinogenesis of gallbladder, and has targeted therapeutic potential.

LncRNA-PAGBC acts as a microRNA sponge and promotes gallbladder tumorigenesis.

Long noncoding RNAs (lncRNAs) play roles in the development and progression of many cancers; however, the contributions of lncRNAs to human gallbladder cancer (GBC) remain largely unknown. In this study, we identify a group of differentially expressed lncRNAs in human GBC tissues, including prognosis-associated gallbladder cancer lncRNA (lncRNA-PAGBC), which we find to be an independent prognostic marker in GBC Functional analysis indicates that lncRNA-PAGBC promotes tumour growth and metastasis of GBC cells. More importantly, as a competitive endogenous RNA (ceRNA), lncRNA-PAGBC competitively binds to the tumour suppressive microRNAs miR-133b and miR-511. This competitive role of lncRNA-PAGBC is required for its ability to promote tumour growth and metastasis and to activate the AKT/mTOR pathway. Moreover, lncRNA-PAGBC interacts with polyadenylate binding protein cytoplasmic 1 (PABPC1) and is stabilized by this interaction. This work provides novel insight on the molecular pathogenesis of GBC.

Gallbladder cancer epidemiology, pathogenesis and molecular genetics: Recent update.

Gallbladder cancer is a malignancy of biliary tract which is infrequent in developed countries but common in some specific geographical regions of developing countries. Late diagnosis and deprived prognosis are major problems for treatment of gallbladder carcinoma. The dramatic associations of this orphan cancer with various genetic and environmental factors are responsible for its poorly defined pathogenesis. An understanding to the relationship between epidemiology, molecular genetics and pathogenesis of gallbladder cancer can add new insights to its undetermined pathophysiology. Present review article provides a recent update regarding epidemiology, pathogenesis, and molecular genetics of gallbladder cancer. We systematically reviewed published literature on gallbladder cancer from online search engine PubMed (http://www.ncbi.nlm.nih.gov/pubmed). Various keywords used for retrieval of articles were Gallbladder, cancer Epidemiology, molecular genetics and bullion operators like AND, OR, NOT. Cross references were manually searched from various online search engines (http://www.ncbi.nlm.nih.gov/pubmed,https://scholar.google.co.in/, http://www.medline.com/home.jsp). Most of the articles published from 1982 to 2015 in peer reviewed journals have been included in this review.

Long non-coding RNA H19 regulates FOXM1 expression by competitively binding endogenous miR-342-3p in gallbladder cancer.

BackgroundLong non-coding RNA (lncRNA) H19 has been reported to involve in many kinds of human cancers and functions as an oncogene. Our previous study found that H19 was over-expressed in gallbladder cancer (GBC) and was shown to promote tumor development in GBC. However, the competing endogenous RNA (ceRNA) regulatory network involving H19 in GBC progression has not been fully elucidated. We aim to detect the role of H19 as a ceRNA in GBC.Methods and ResultsIn this study, the expression of H19 and miR-342-3p were analyzed in 35 GBC tissues and matched normal tissues by using quantitative polymerase chain reaction (qRT-PCR). We demonstrated H19 was overexpressed and negatively correlated with miR-342-3p in GBC. By dual-luciferase reporter assays, RNA-binding protein immunoprecipitation (RIP) and RNA pull-down assays, we verified that H19 was identified as a direct target of miR-342-3p. QRT-PCR and Western-blotting assays demonstrated that H19 silencing down-regulated, whereas over-expression enhanced the expression of miR-342-3p targeting FOXM1 through competitively ‘sponging’ miR-342-3p. Furthermore, transwell invasion assays and cell cycle assays indicated that H19 knockdown inhibited both cells invasion and proliferation, but this effects was attenuated by co-transfection of siRNA-H19 and miR-342-3p inhibitor in GBC cells. In vivo, tumor volumes were decreased significantly in H19 silenced group compared to the control group, but was attenuated by co-transfection of shRNA-H19 and miR-342-3p inhibitor, which were stablely constructed through lenti-virus vector.ConclusionOur results suggest a potential ceRNA regulatory network involving H19 regulates FOXM1 expression by competitively binding endogenous miR-342-3p in GBC. This mechanism may contribute to a better understanding of GBC pathogenesis and provides potential therapeutic strategy for GBC.

Abstract 3441: Associations between lipopolysaccharide (LPS) and LPS pathway biomarkers and gallbladder cancer are modulated by markers of systemic inflammation

Abstract Little is known about gallbladder cancer (GBC) etiology. Gallstones (GS) are a critical factor in GBC pathogenesis. Bacterial infections, such as Salmonella Typhi and Helicobacter pylori, also may contribute to the development of GBC. We previously observed strong associations between inflammation markers and GBC vs. GS patients and hypothesized that bacterial infections may explain in part associations between GBC and inflammation. In the current study of 41 GBC cases and 124 GS patients from Shanghai, China, we evaluated relationships between GBC and the bacterial infection-related proteins plasma lipopolysaccharide (LPS) and two LPS-pathway proteins, soluble CD14 (sCD14) and lipid binding protein (LBP), as measured by ELISA. Further, we explored the extent to which these associations were mediated through systemic inflammation. Logistic regression was conducted to assess associations of GBC with plasma LPS, sCD14, and LBP levels. To evaluate mediation through inflammation, an inflammation score was calculated by summing the value of 15 previously measured GBC-associated serum inflammation markers (β-coefficient multiplied by 1 if above the median or 0 if below the median) and included in the models. sCD14 and LBP levels (above vs. below the median) were strongly associated with GBC [Odds Ratio (95% Confidence Interval): 2.75 (1.27-6.29) for sCD14 and 2.83 (1.32-6.46) for LBP]. The highest vs. lowest tertiles of sCD14 and LBP were associated with a five-fold increased risk of GBC. These associations were attenuated when the inflammation score was entered into the model. Plasma LPS demonstrated a borderline association after adjustment for inflammation score [OR (95% CI): 2.46 (0.96-6.56)]. In contrast, the ORs for inflammation score remained >20 after adjusting for each bacteria-related marker. These findings suggest that bacterial infections may contribute to GBC etiopathogenesis through inflammatory pathways. LPS, sCD14, and LBP and Gallbladder Cancer Relative to Gallstone ControlsLPSSex & Age Adjusted OR (95% CI)Sex, Age, & Inflammation Score Adjusted OR (95% CI)Detectable Level1.001.00Undetectable Level1.81 (0.85-3.80)2.46 (0.96-6.56)sCD14Sex, Age, & Batch Adjusted OR (95% CI)Sex, Age, Batch, & Inflammation Score Adjusted OR (95% CI)Below Median1.001.00Above Median2.75 (1.27-6.29)1.05 (0.39-2.83)1st Tertile1.001.002nd Tertile1.42 (0.45-4.71)0.44 (0.10-1.80)3rd Tertile5.27 (1.98-16.06)1.37 (0.38-5.04)p-trend0.00060.25LBPSex, Age, & Batch Adjusted OR (95% CI)Sex, Age, Batch, & Inflammation Score Adjusted OR (95% CI)Below Median1.001.00Above Median2.83 (1.32-6.46)1.63 (0.64-4.28)1st Tertile1.001.002nd Tertile1.95 (0.61-6.99)1.53 (0.37-7.01)3rd Tertile5.01 (1.84-16.25)2.09 (0.58-8.71)p-trend0.00080.25 Citation Format: Alison L. Van Dyke, Troy J. Kemp, Amanda F. Corbel, Bin Zhu, Yu-Tang Gao, Bingsheng Wang, Asif Rashid, Ming-Chang Shen, Allan Hildesheim, Ann W. Hsing, Ligia A. Pinto, Jill Koshiol. Associations between lipopolysaccharide (LPS) and LPS pathway biomarkers and gallbladder cancer are modulated by markers of systemic inflammation. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3441.

RNA sequencing-based analysis of gallbladder cancer reveals the importance of the liver X receptor and lipid metabolism in gallbladder cancer.

Gallbladder cancer (GBC) is an aggressive malignancy. Although surgical resection may be curable, most patients are diagnosed at an advanced unresectable disease stage. Cholelithiasis is the major risk factor; however the pathogenesis of the disease, from gallstone cholecystitis to cancer, is still not understood. To understand the molecular genetic underpinnings of this cancer and explore novel therapeutic targets for GBC, we examined the key genes and pathways involved in GBC using RNA sequencing. We performed gene expression analysis of 32 cases of surgically-resected GBC along with normal gallbladder tissue controls. We observed that 519 genes were differentially expressed between GBC and normal GB mucosal controls. The liver X receptor (LXR)/retinoid X receptor (RXR) and farnesoid X receptor (FXR) /RXR pathways were the top canonical pathways involved in GBC. Key genes in these pathways, including SERPINB3 and KLK1, were overexpressed in GBC, especially in female GBC patients. Additionally, ApoA1 gene expression suppressed in GBC as compared with normal control tissues. LXR and FXR genes, known to be important in lipid metabolism also function as tumor suppressors and their down regulation appears to be critical for GBC pathogenesis. LXR agonists may have therapeutic value and as potential therapeutic targets.

IL17 producing γδT cells induce angiogenesis and are associated with poor survival in gallbladder cancer patients.

Despite conventional treatment modalities, gallbladder cancer (GBC) remains a highly lethal malignancy. Prognostic biomarkers and effective adjuvant immunotherapy for GBC are not available. In the recent past, immunotherapeutic approaches targeting tumor associated inflammation have gained importance but the mediators of inflammatory circuit remain unexplored in GBC patients. In the current prospective study, we investigated the role of IL17 producing TCRγδ(+) (Tγδ17), CD4(+) (Th17), CD8(+) (Tc17) and regulatory T cells (Tregs) in pathogenesis of GBC. Analysis by multi-color flow cytometry revealed that compared to healthy individuals (HI), Tγδ17, Th17 and Tc17 cells were increased in peripheral blood mononuclear cells (PBMCs) and tumor infiltrating lymphocytes (TIL) of GBC patients. Tregs were decreased in PBMCs but increased in TILs of GBC patients. The suppressive potential of Tregs from GBC patients and HI were comparable. Serum cytokines profile of GBC patients showed elevated levels of cytokines (IL6, IL23 and IL1β) required for polarization and/or stabilization of IL17 producing cells. We demonstrated that Tγδ17 cells migrate toward tumor bed using CXCL9-CXCR3 axis. IL17 secreted by Tγδ17 induced productions of vascular endothelial growth factor and other angiogenesis related factors in GBC cells. Tγδ17 cells promote vasculogenesis as studied by chick chorioallantoic membrane assay. Survival analysis showed that Tγδ17, Th17 and Treg cells in peripheral blood were associated with poor survival of GBC patients. Our findings suggest that Tγδ17 is a protumorigenic subtype of γδT cells which induces angiogenesis. Tγδ17 may be considered as a predictive biomarker in GBC thus opening avenues for targeted therapies.

MicroRNA aberrations: An emerging field for gallbladder cancer management.

Gallbladder cancer (GBC) is infrequent but most lethal biliary tract malignancy characterized by an advanced stage diagnosis and poor survival rates attributed to absence of specific symptoms and effective treatment options. These necessitate development of early prognostic/predictive markers and novel therapeutic interventions. MicroRNAs (miRNAs) are small, non-coding RNA molecules that play a key role in tumor biology by functioning like tumor suppressor- or onco- genes and their aberrant expression are associated with the pathogenesis of several neoplasms with overwhelming clinical implications. Since miRNA signature is tissue specific, here, we focused on current data concerning the miRNAs aberrations in GBC pathogenesis. In GBC, miRNAs with tumor suppressor activity (miR-135-5p, miR-335, miR-34a, miR-26a, miR-146b-5p, Mir-218-5p, miR-1, miR-145, mir-130a) were found downregulated, while those with oncogenic property (miR-20a, miR-182, mir-155) were upregulated. The expression profile of miRNAs was significantly associated with GBC prognosis and prediction, and forced over-expression/ inhibition of these miRNAs was shown to affect tumor growth and development. Further, differential expression of miRNAs in the blood samples of GBC patients suggest miRNAs as promising noninvasive biomarker. Thus, miRNAs represent potential candidate for GBC management, though many hurdles need to be overcome before miRNAs therapy can be clinically applied to GBC prevention and treatment.

Epigenetic regulation of APC in the molecular pathogenesis of gallbladder cancer.

Background & objectives:Loss of function of adenomatous polyposis coli (APC) has been reported in cancer. The two promoters of APC, 1A and 1B also have roles in cancer. But, the epigenetic role of APC promoters is not yet clear in gallbladder cancer (GBC) and gallstone diseases (GSD). We undertook this study to determine the epigenetic role of APC in GBC and GSD.Methods:Methylation-specific (MS)-PCR was used to analyze the methylation of APC gene. The expression of APC gene was studied by semi-quantitative PCR, real-time PCR and immunohistochemistry (IHC) in GBC, GSD and adjacent normal tissues.Results:Of the two promoters, APC 1A promoter was found methylated in 96 per cent GBC (P=0.0155) and 80 per cent GSD (P=0.015). Exon 1 was downregulated in grade II (P=0.002) and grade III (P=0.0001) of GBC, while exon 2 was normally expressed. Scoring analysis of IHC revealed 0 or negativity in 34.48 per cent (P=0.057) and 1+ in 24.14 per cent (P=0.005) GBC cases suggesting loss of APC expression.Interpretation & conclusions:The present findings indicate epigenetic silencing of APC in advanced GBC. The methylation pattern, followed by expression analysis of APC may be suggested for diagnostic, prognostic and therapeutic purposes in GBC in future.