Gastrointestinal stromal tumors (GISTs) of the gallbladder are representative of an extremely rare group of tumors. We have encountered a patient with a malignant GIST of the gallbladder and presented it with a review of some articles. A 72-yr-old woman initially presented with right upper quadrant abdominal pain, fever and chills. Emergency cholecystectomy was performed under the impression of gallbladder empyema. Liver metastasis was found at 7 months postoperatively and the patient expired 9 months after the surgery. At the time of cholecystectomy, the gallbladder showed a necrotic serosal surface with an irregular thickened wall. A mass, 6 cm in length and 3 cm in width, encircled the whole wall of the neck and upper body of the gallbladder. Microscopic findings revealed frequent mitotic figures (more than 20/50 HPF) and tumor necrosis. Hyperchromatic, pleomorphic and spindle shaped neoplastic cells that were arranged in a pattern of short fascicles infiltrated the entire layer of the gallbladder. The tumor cells were immunoreactive for CD117 antigen (c-kit protein) and vimentin. They were negative for desmin, smooth muscle actin and S-100 protein. Mutations of the c-kit proto-oncogene were not found in this case. These findings were sufficient to provide enough clinical, histopathological and immunohistochemicalevidence in diagnosing our case as a malignant GIST.
Aged ; Female ; Gallbladder Neoplasms/genetics/metabolism/*pathology ; Gastrointestinal Stromal Tumors/genetics/metabolism/*pathology ; Humans ; Proto-Oncogene Protein c-kit/genetics/metabolism

CA-19-9 Antigen ; metabolism ; Carcinoembryonic Antigen ; metabolism ; Carcinoma ; pathology ; Carcinoma in Situ ; etiology ; genetics ; metabolism ; pathology ; Chromosomes, Human, Pair 17 ; Chromosomes, Human, Pair 5 ; Diagnosis, Differential ; Gallbladder Diseases ; pathology ; Gallbladder Neoplasms ; etiology ; genetics ; metabolism ; pathology ; Humans ; Loss of Heterozygosity ; Microsatellite Instability ; Polyps ; pathology ; Precancerous Conditions ; etiology ; genetics ; metabolism ; pathology

Antigens ; immunology ; Antigens, Neoplasm ; drug effects ; immunology ; metabolism ; Gallbladder Neoplasms ; immunology ; metabolism ; pathology ; Gene Expression Regulation, Neoplastic ; Humans ; Male ; Neoplastic Stem Cells ; immunology ; Octamer Transcription Factor-3 ; genetics ; metabolism ; Prostatic Neoplasms ; genetics ; metabolism ; pathology ; Tumor Cells, Cultured

OBJECTIVE: To examine the expressive level of enhancer of zesle homolog 2 (EZH2) and phosphatase and tension homolog (PTEN), and to explore its clinicopathological significance in benign and malignant lesion of gallbladder. METHODS: EnVision immunohistochemical method was used to detect the expressive levels of EZH2 and PTEN in routinely paraffin-embedded sections in the resected specimens of gallbladder adenocarcinoma (n = 108), peritumoral tissues (n = 46), adenomatous polyp(n = 15), and chronic cholecystitis (n = 35). RESULTS: The positive rate of EZH2 was significantly higher in gallbladder adenocarcinoma than that in peritumoral tissues (chi(2) = 24.49, P < 0.01), adenomatous polyp(chi(2) = 11.68, P < 0.01), and chronic cholecystitis (chi(2) = 31.62, P < 0.01). The benign lesions in the positive cases of EZH2 and (or) the negative ones of PTEN showed the moderately- or severely- atypical hyperplasia of gallbladder epithelium. The positive rate of PTEN was significantly lower in gallbladder adenocarcinoma than that in peritumoral tissues(n = 20.20, P < 0.01), adenomatous polyp(chi(2)=10.81, P<0.01), and chronic cholecystitis (n = 29.83, P < 0.01).The positive rates of EZH2 were significantly lower in the highly-differentiated adenocarcinoma, the maximal diameter of mass < 2 cm, non-metastasis of lymphnodes, and non-infiltration of regional tissues than those in the moderately or low-differentiated adenocarcinoma, the maximal diameter > or = 2 cm, metastasis of lymphnode, and infiltration of regional tissues (P < 0.05 or P < 0.01). High inconsistency was found between the expression of EZH2 and PTEN in gallbladder adenocarcinoma (P < 0.05). CONCLUSION Expression of EZH2 and/or PTEN might be important biological markers in the carcinogenesis, progression, biological behaviors and prognosis of gallbladder adenocarcinoma.
Adenocarcinoma ; genetics ; pathology ; Adult ; Biomarkers, Tumor ; DNA-Binding Proteins ; biosynthesis ; Enhancer of Zeste Homolog 2 Protein ; Female ; Gallbladder Neoplasms ; genetics ; metabolism ; Gene Expression Regulation, Neoplastic ; Humans ; Male ; Middle Aged ; PTEN Phosphohydrolase ; biosynthesis ; Polycomb Repressive Complex 2 ; Prognosis ; Transcription Factors ; biosynthesis

BACKGROUNDThe precise molecular mechanisms underlying the gallbladder carcinoma (GBC) metastasis has not been fully elucidated.

METHODSIn the present study, metastasis-associated proteins were identified by comparative proteomic analysis. The functional study of the candidate protein vimentin was further investigated. First, a pair of higher and lower metastatic sublines (termed GBC-SD/M3 and GBC-SD, respectively), originated from the same parental cell line, was screened by spontaneous tumorigenicity and metastasis in vivo in animal study and further characterized by metastatic phenotypes analysis in vitro. Subsequently, a proteomic approach comprised two-dimensional gel electrophoresis analysis and mass spectroscopy was used to identify and compare the protein expression patterns between higher metastatic GBC-SD/M3 and lower metastatic GBC-SD cell lines. Then twenty-six proteins were identified.

RESULTSAmong the 26 proteins identified, fourteen proteins were up-regulated and 12 proteins were down-regulated in GBC-SD/M3. Vimentin was identified and found to be overexpressed in GBC-SD/M3 as compared with GBC-SD. This result was further confirmed by quantitative PCR and Western blotting analysis. Furthermore, the cell migration and invasion potency of GBC-SD/M3 in vitro was remarkably suppressed after small interference RNA-mediated knockdown of vimentin. Moreover, immunoblot and immunohistochemical analysis on 12 human GBC specimens showed consistently increased vimentin expression in metastases compared with primary tumors.

CONCLUSIONTumor vimentin level may reflect the pathological progression in some GBC and may be a useful marker for predicting tumor metastasis and a therapeutic target for the treatment of GBC patients with metastases.

Animals ; Blotting, Western ; Cell Line, Tumor ; Cell Movement ; genetics ; physiology ; Electrophoresis, Gel, Two-Dimensional ; Gallbladder Neoplasms ; genetics ; metabolism ; pathology ; Humans ; Immunohistochemistry ; Mice ; Mice, Nude ; Neoplasm Metastasis ; genetics ; pathology ; RNA Interference ; Reverse Transcriptase Polymerase Chain Reaction ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; Vimentin ; genetics ; metabolism

OBJECTIVETo examine the expression of tissue factor pathway inhibitor-2 (TFPI-2) in gallbladder cancer (GBC) and to investigate the anti-cancer activities of TFPI-2 against the growth of GBC.

METHODSTFPI-2 expression in gallbladder normal tissues, gallbladder polyp (GBP) tissues and GBC tissues were examined by reverse transcriptase polymerase chain reaction (RT-PCR), Western blot and immunohistochemical staining. Adenovirus carrying human TFPI-2 gene (Ad5-TFPI-2) were constructed and its anti-cancer effects were investigated in xenograft tumors. Xenograft tumors were constructed by injection of GBC-SD and SGC-996 cells into the flank of nude mice and the volume of xenograft tumors was measured every 3 days until the sacrifice of mice. The apoptosis index of xenograft tumors was examined by TUNEL assay. The status of Bax, Bcl-2 and caspase-3 was examined by Western blot assay.

RESULTSTFPI-2 expression was profoundly lower in GBC tissues (87.0%) when compared to normal tissues (23.3%) and GBP tissues (52.2%; χ(2) = 21.104, P = 0.000). Ad-TFPI-2 significantly inhibited the growth of xenograft tumors in nude mice. Ad-TFPI-2 inhibited GBC-SD cell growth through the induction of apoptosis. The means of total apoptotic cells per field were much higher in Ad5-TFPI-2 group than those in PBS and Ad5-GFP groups. Ad5-TFPI-2 elevated the expression of Bax and cleaved caspase-3, while it decreased the expression of Bcl-2.

CONCLUSIONSTFPI-2 gene and protein was down-regulated in GBC and the down-regulation of TFPI-2 may play a role in the tumorigenesis of GBC. Adenovirus-mediated TFPI-2 can inhibit GBC growth through the induction of apoptosis.

Adenoviridae ; genetics ; Aged ; Animals ; Apoptosis ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Female ; Gallbladder Neoplasms ; metabolism ; pathology ; therapy ; Genetic Therapy ; Glycoproteins ; genetics ; metabolism ; Humans ; Male ; Mice ; Mice, Nude ; Middle Aged ; Neoplasm Transplantation ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; bcl-2-Associated X Protein ; metabolism

OBJECTIVETo explore the anti-tumor mechanism of norcantharidin (NCTD) for the implanted tumors of human gallbladder carcinoma in nude mice in vivo.

METHODSAnimal model of implanted tumors of human gallbladder carcinoma in nude mice was established. Mice were randomly divided into control, 5-FU, NCTD and NCTD + 5-FU groups and were taken different treatment. The expressions of PCNA, Ki-67, cyclin D1, p27, Bcl-2, Bax, Survivin, nm23/nm23-H1, MMP2 and TIMP2 proteins or genes in each tissue section of every group were determined by immunohistochemistry and RT-PCR.

RESULTS(1) On proliferation-related gene proteins, the expression of PCNA, Ki-67, cyclin D1 was significantly decreased, with significantly increased expression of p27 protein, in paraffin sections of NCTD group when compared with control group (P < 0.05); The expression of PCNA mRNA, cyclin D1 mRNA was decreased, with significantly increased expression of p27 mRNA in NCTD group. (2) On apoptosis-related gene proteins, the expression of Bcl-2 was significantly decreased in paraffin sections of NCTD group when compared with control group (P < 0.05); The expression of Bcl-2 mRNA, Survivin mRNA was significantly decreased, with significantly increased expression of Bax mRNA in NCTD group. (3) There was significant difference on invasion around tumor and lung metastasis in NCTD group when compared with control group (P < 0.01). On metastasis-related gene proteins, the expression of nm23 and TIMP2 was significantly increased, with significantly decreased expression of MMP2 in paraffin sections of NCTD group when compared with control group (P < 0.05); The expression of nm23-H1 mRNA, TIMP2 mRNA was significantly increased, with significantly decreased expression of MMP2 mRNA in NCTD group.

CONCLUSIONSThe anti-tumor mechanism of NCTD for human gallbladder carcinoma in nude mice might correlated with inhibition of cell proliferation, blockage of cell cycle, induction of cell apoptosis, reducing of cell motility and invasive capability, alteration of the expression of proliferation-, apoptosis- and metastasis-related gene proteins such as PCNA, Ki-67, cyclin D1, p27, Bcl-2, Bax, Survivin, nm23, MMP2 and TIMP2.

Animals ; Apoptosis ; drug effects ; Bridged Bicyclo Compounds, Heterocyclic ; pharmacology ; Cell Proliferation ; drug effects ; Cyclin D1 ; biosynthesis ; genetics ; Gallbladder Neoplasms ; drug therapy ; metabolism ; pathology ; Humans ; Ki-67 Antigen ; biosynthesis ; genetics ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Proliferating Cell Nuclear Antigen ; biosynthesis ; genetics ; RNA, Messenger ; genetics ; bcl-2-Associated X Protein ; biosynthesis ; genetics