OBJECTIVETo research the effects of bile acids on the expression of interleukin-6 (IL-6) and the cell viability in QBC939 cell line.

METHODSHuman cholangiocarcinoma cells were stimulated with 800 µmol/L bile acid (CA), 100 µmol/L deoxycholate (DCA), 100 µmol/L chenodeoxycholic acid (CDCA), 1200 µmol/L gly acid (GCA), 200 µmol/L glycodeoxycholic acid (GDCA) and 300 µmol/L gly chenodeoxycholic acid (GCDCA).MTT assay and ELISA were used to detect the cell viability and the expression of IL-6 at 24 h, 48 h and 72 h.

RESULTSTreated by DCA, CDCA and GCDCA for 48 hours, the cell viability ratios changed to 0.61, 0.58 and 1.26, which were significant differences between control group and treated groups. And after 72 hours, the viability ratios of group CA, group DCA, group CDCA, group GCA, group GDCA and group GCDCA turned into 0.48, 0.50, 0.42, 1.29, 1.30 and 1.41. The differences of cell viability between bile acid-treated groups and control group were significant (P < 0.05). The expression of IL-6 in control group at 48 h and 72 h was (198 ± 32) ng/L and (323 ± 34) ng/L, while treated by CA, DCA, CDCA, GCA, GDCA and GCDCA respectively for 48 hours, the expression of IL-6 altered to (106 ± 33) ng/L, (88 ± 29) ng/L, (116 ± 54) ng/L, (413 ± 21) ng/L, (587 ± 32) ng/L and (366 ± 30) ng/L. After 72 hours, the expression of IL-6 of each bile acid-treated groups as above was (123 ± 66) ng/L, (45 ± 21) ng/L, (74 ± 45) ng/L, (792 ± 13) ng/L, (1310 ± 22) ng/L and (845 ± 18) ng/L, respectively. The differences between each bile acid-treated group and control group were significant (P < 0.05).

CONCLUSIONSFree bile acids (CA, DCA and CDCA) can inhibit the expression of IL-6 and the cell viability, while glycine conjugates (GCA, GDCA and GCDCA) can promote the expression of IL-6 and the cell viability. Bile acids can change tumor cell viability via IL-6 pathway.

Bile Acids and Salts ; pharmacology ; Bile Duct Neoplasms ; metabolism ; pathology ; Cell Line, Tumor ; Cell Survival ; drug effects ; Humans ; Interleukin-6 ; metabolism

OBJECTIVETo study the correlation between clinicopathological features and serum carbohydrate antigen 19-9 (CA19-9)/carcinoembryonic antigen (CEA) in patients with extrahepatic cholangiocarcinoma (ECC).

METHODSThe clinicopathological data of 126 cases of extrahepatic cholangiocarcinoma treated in our department from Jan. 1999 to Dec. 2012 were collected and analyzed in this study. The correlation between clinicopathological features and sensitivity of CA19-9/CEA was analyzed by chi-square test. The correlation of clinicopathological features and value of serum CA19-9/CEA was analyzed by t test and F test.

RESULTSThe average value of CA19-9 before surgery in the 126 patients was 595.3 U/ml. The values of CA19-9 in 91 patients were abnormal and the sensitivity of CA19-9 was 72.2%. The average value of CEA before surgery was 12.6 U/ml. The value of CEA in 26 patients were abnormal and the sensitivity of CEA was 20.6%. The values of combined detection of serum CA19-9 and CEA before surgery were abnormal in a total of 97 cases with a sensitivity of 77.0%. There was no significant correlation between clinicopathological features and sensitivity of CA19-9 (P > 0.05). The location of tumor was significantly correlated to the diagnostic sensitivity of CEA. The sensitivity of CEA to distal ECC was only 15.4%. The value of CA19-9 was relatively high in patients >60-year old or with neural invasion, while CEA was higher when tumor was located in the middle of bile duct (P < 0.05). There was no significant difference of serum CA19-9 before and after jaundice reduction (P > 0.05).

CONCLUSIONSThe diagnostic sensitivity of CA19-9 is not affected by gender, age, blood type, tumor location, degree of differentiation, tumor size, T stage, vascular tumor thrombus, lymph node metastasis, perineural invasion, and preoperative jaundice. However, the diagnostic sensitivity of CEA is affected by tumor location. The value of CA19-9 is correlated with tumor invasion and is relatively high in patients above 60 years old.

Bile Duct Neoplasms ; metabolism ; pathology ; Bile Ducts, Intrahepatic ; metabolism ; pathology ; Biomarkers, Tumor ; metabolism ; CA-19-9 Antigen ; metabolism ; Carcinoembryonic Antigen ; metabolism ; Cholangiocarcinoma ; metabolism ; pathology ; Humans ; Lymphatic Metastasis

No abstract available.
Aged ; Bile Duct Neoplasms/metabolism/*pathology ; Bile Ducts, Intrahepatic/metabolism/*pathology ; Cholangiocarcinoma/metabolism/*pathology ; Humans ; Immunohistochemistry ; Keratin-19/metabolism ; Male ; Positron-Emission Tomography ; Tomography, X-Ray Computed

Actins ; metabolism ; Antigens, CD ; metabolism ; Antigens, Differentiation, Myelomonocytic ; metabolism ; Bile Duct Neoplasms ; metabolism ; pathology ; surgery ; Bile Ducts, Intrahepatic ; Cholangiocarcinoma ; metabolism ; pathology ; surgery ; Female ; Giant Cells ; metabolism ; pathology ; Humans ; Keratins ; metabolism ; Liver Neoplasms ; metabolism ; pathology ; surgery ; Osteoclasts ; metabolism ; pathology ; Vimentin ; metabolism

OBJECTIVETo clarify the relationship between the loss of expression of the deleted in pancreatic carcinoma locus 4 (DPC4) proteins and the pathogenesis of biliary tract carcinoma.

METHODS71 primary biliary tract carcinoma (BTCa), including 38 common bile duct (CBD) carcinomas, 18 gallbladder carcinomas, 15 hilar bile ducts (HBD) carcinomas were examined by immunohistochemical staining. In addition, the CBD carcinomas were divided into two groups: tumors with metastasis (M(+) group, 27 cases) and tumors without metastasis (M(-) group, 11 cases).

RESULTSThe frequency of loss of the expression of DPC4 protein was 32.8% in BTCa, 47.3% in CBD carcinoma, 11% in gallbladder carcinoma, 13% in HBD carcinoma. Comparison of the frequency of loss expression of DPC4 was significant statistical difference in CBD carcinoma versus gallbladder carcinoma and HBD carcinoma (P < 0.01). The frequency of loss expression of DPC4 was 48.1% in the M(+) group and 45.4% in the M(-) group.

CONCLUSIONThere are a close relationship between pathogenesis of BTCa and inactivation of DPC4 and different frequencies of DPC4 gene alternation in various locations of the biliary tract, which are not significantly increased with tumor metastasis in BTCa.

Bile Duct Neoplasms ; Biliary Tract ; Carcinoma ; DNA-Binding Proteins ; metabolism ; Humans ; Pancreatic Neoplasms ; Smad4 Protein ; Trans-Activators

There has been no report for the expression of cyclooxygenase-2 (COX-2) and its clinicopathologic and biologic significance in ampulla of Vater cancer. This study was aimed for the clarification of COX-2 expression and its biologic roles in ampulla of Vater cancer. Fourty-six patients with ampulla of Vater cancer were enrolled and their COX-2 expression and clinicopathologic features were analyzed. The median age of patients was 60 yr and the mean duration of follow-up was 35 months (range: 14-82 months). Immunohistochemical stainings for COX-2, Ki-67, CD34 and TUNEL staining were performed. The immunoreactive COX-2 expression was present in 24 (52.2%) patients of ampulla of Vater cancer and mainly localized in cytosolic and perinuclear region. There was no significant difference in the length of survival between COX-2 postive and negative group (p=0.9420 by Log Rank test). Also, there were no significant differences of proliferation index (p=0.326), apoptotic index (p=0.764) and microvessel density (p=0.135) between COX-2 positive and negative group. Initial pTNM stage (p=0.0028 by Log Rank test) and blood transfusion over 4 pints during operation (p=0.0254 by Log Rank test) were independent prognostic factor in patients with ampulla of Vater cancer. It is suggested that immunoreactivity of COX-2 is not correlated with clinicopathologic and biologic features of ampulla of Vater cancer.
Adult ; Aged ; Ampulla of Vater*/enzymology ; Ampulla of Vater*/pathology ; Common Bile Duct Neoplasms/enzymology* ; Common Bile Duct Neoplasms/pathology* ; Female ; Human ; Immunoenzyme Techniques ; Isoenzymes/metabolism* ; Male ; Middle Aged ; Prostaglandin-Endoperoxide Synthase/metabolism* ; Statistics ; Survival Rate

BACKGROUND/AIMS: Hedgehog protein is an essential molecule for gastrointestinal tract development, and disruption of hedgehog signaling pathway is linked to some gastrointestinal tumorigenesis. Here, we performed hedgehog immunostaining in periampullary cancer to evaluate the differences according to the location type of cancer and the differentiation of adenocarcinoma. METHODS: We retrieved surgical specimens from 43 periampullary cancer patients (15 ampulla of Vater cancer, 12 distal common bile duct cancer, 13 pancreatic head cancer, and 3 combined ampulla of Vater/bile duct cancer). Immunohistochemical stain was performed in both normal and cancerous tissue portions of each case using Sonic hedgehog (H-160) rabbit polyclonal antibody. Immunohistochemical stain results were grouped into three groups according to the percentage of positive cytoplasmic stain in tumor volume (unstained: <5%, weakly stained: 5-50%, and strongly stained: >50%). RESULTS: All of the normal tissue revealed negative immunohistochemical stain while cancerous tissue revealed positivity in 95.3% (41/43 cases). Strongly stained cases were more frequently seen in ampulla of Vater cancers (13/15) and in combined ampulla of Vater/bile duct cancers (3/3) than in distal common bile duct cancers (4/12) and in pancreatic head cancers (3/13) (p=0.002). In addition, strongly stained cases were more frequently seen in well-differentiated adenocarcinoma than the others (p<0.001). CONCLUSIONS: Most of the periampullary cancers show hedgehog protein expression. In addition, hedgehog protein immunostainings shows stronger expression in ampulla of Vater cancers and in well-differentiated adenocarcinoma.
Adenocarcinoma/metabolism ; Aged ; *Ampulla of Vater ; Common Bile Duct Neoplasms/*metabolism ; English Abstract ; Female ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Pancreatic Neoplasms/*metabolism ; Trans-Activators/*metabolism

OBJECTIVETo compare the immunoprofiles of intrahepatic cholangiocarcinoma and metastatic colorectal adenocarcinoma for mucin glycoproteins (including MUC1, MUC2, MUC5AC and MUC6) and cytokeratins (including CK7, CK19 and CK20), and to assess their diagnostic value.

METHODSOne hundred cases of intrahepatic cholangiocarcinoma and 21 cases of metastatic colorectal adenocarcinoma were enrolled into the study. Immunohistochemical study for MUC1, MUC2, MUC5AC, MUC6, CK7, CK19 and CK20 was carried out in all cases by EnVision method.

RESULTSIn intrahepatic cholangiocarcinoma, the expression rates of MUC1, MUC2, MUC5AC and MUC6 were 61.0%, 2.0%, 22.0% and 8.0% respectively, as compared to 57.1%, 47.6%, 19.0% and 23.8% respectively in metastatic colorectal adenocarcinoma. On the other hand, the expression rates of CK7, CK19 and CK20 in intrahepatic cholangiocarcinoma were 73.0%, 53.0% and 15.0% respectively, in contrast to 14.3%, 90.5% and 85.7% respectively in metastatic colorectal adenocarcinoma. The difference in expressions of MUC2, MUC6, CK7 and CK20 carried statistical significance.

CONCLUSIONSThe immunoprofile for mucin glycoproteins and cytokeratins provides important clues in distinguishing between intrahepatic cholangiocarcinoma and metastatic colorectal adenocarcinoma to liver. The immunophenotype of MUC2-/MUC6-/CK7+/CK20- indicates the diagnosis of intrahepatic cholangiocarcinoma, while MUC2+/MUC6+/CK7-/CK20+ suggests the possibility of metastatic colorectal adenocarcinoma.

Adenocarcinoma ; metabolism ; pathology ; Aged ; Bile Duct Neoplasms ; genetics ; metabolism ; pathology ; Bile Ducts, Intrahepatic ; pathology ; Biomarkers, Tumor ; analysis ; Cholangiocarcinoma ; genetics ; metabolism ; pathology ; Colorectal Neoplasms ; metabolism ; pathology ; Female ; Glycoproteins ; metabolism ; Humans ; Keratins ; metabolism ; Male ; Middle Aged ; Mucins ; metabolism ; Neoplasm Staging ; classification

Objective To investigate the expression and the potential roles of long non-coding RNA(lncRNA)cancer susceptibility candidate 2(CASC2)and imprinted gene H19 in extrahepatic cholangiocarcinoma(ECC). Methods Four samples from patients with ECC were collected for high-throughput sequencing which was conducted to reveal the transcriptomic profiles of lncRNA CASC2 and H19.Bioinformatics tools were employed to predict the potential roles of the two genes.Another 22 ECC tissue samples and the cholangiocarcinoma cell lines(RBE,QBC939,HuH-28,and HuCCT1)with different degrees of differentiation were selected for validation.The para-carcinoma tissue and normal human intrahepatic biliary epithelial cell(HIBEC)were used as the control groups.The expression levels of lncRNA CASC2 and H19 in carcinoma tissue,para-carcinoma tissue,and cell lines were determined by real-time quantitative polymerase chain reaction(qRT-PCR).The correlation analysis was carried out for the clinical indicators of patients with the expression levels of the target genes. Results The two target genes showed significantly different expression between carcinoma tissue and para-carcinoma tissue(all P<0.05).Specifically,CASC2 had higher expression level in the carcinoma tissue than in the para-carcinoma tissue(t=1.262,P=0.025),whereas the expression of H19 showed an opposite trend(t=1.285,P=0.005).The expression levels of CASC2 in QBC939(t=8.114,P=0.015)and HuH-28(t=9.202,P=0.012)cells were significantly higher than that in the control group.The expression levels of H19 were significantly lower in RBE(t=-10.244,P<0.001),QBC939(t=-10.476,P<0.001),HuH-28(t=-19.798,P<0.001),and HuCCT1(t=-16.193,P=0.004)cells than in the control group.Bioinformatics analysis showed that CASC2 was mainly involved in the metabolic process and H19 in the development of multicellular organisms.Both CASC2 and H19 were related to catalytic activity.The expression level of lncRNA CASC2 was correlated with pathological differentiation(χ ²=6.222,P=0.022)and lymph node metastasis(χ²=5.455,P=0.020),and that of lncRNA H19 with pathological differentiation(χ²=1.174,P=0.029)and tumor size(χ²=-0.507,P=0.037). Conclusions In the case of ECC,lncRNA CASC2 and H19 have transcription disorders.lncRNA CASC2 is generally up-regulated in the carcinoma tissue,while H19 is down-regulated.Both genes have the potential to become new molecular markers for ECC.
Bile Duct Neoplasms/genetics* ; Bile Ducts, Intrahepatic/metabolism* ; Cholangiocarcinoma/genetics* ; Gene Expression Regulation, Neoplastic ; Humans ; RNA, Long Noncoding/genetics* ; Tumor Suppressor Proteins/genetics*

Carcinosarcoma ; metabolism ; pathology ; surgery ; Common Bile Duct Neoplasms ; metabolism ; pathology ; surgery ; Diagnosis, Differential ; Female ; Humans ; Immunohistochemistry ; Keratins ; metabolism ; Middle Aged ; Vimentin ; metabolism