OBJECTIVETo study the clinicopathologic features, immunophenotype, histological diagnosis and prognosis of hepatic epithelioid angiomyolipoma.

METHODSClinical data of 25 cases of hepatic epithelioid angiomyolipoma were collected along with follow-up study of the patients. The pathological features were documented and immunohistochemical study of various markers was performed with an emphasis on diagnosis and differential diagnosis.

RESULTSHepatic epithelioid angiomyolipoma was more commonly found in young women without characteristic clinical symptoms. Its morphological features were characterized by marked cytological atypia, relatively rare mitotic figures; radial distribution of tumor cells around the thin-walled blood vessels or muscular vessels; and the presence of common multinucleated giant cells and large ganglion-like tumor cells. The tumor cells expressed both melanoma cell markers (HMB45, MART-1) and smooth muscle cell markers (SMA). Tumor cells expressed various other markers including ER 16% (4/25), PR 32% (8/25), TFE3 24% (6/25) and p53 60% (15/25).

CONCLUSIONSHepatic epithelioid angiomyolipoma has variable morphological features and characteristic immunohistochemical phenotype. The differential diagnoses include a variety of tumors. The biological behavior of the tumor tends to be benign.

Age Factors ; Angiomyolipoma ; genetics ; immunology ; metabolism ; pathology ; Biomarkers, Tumor ; metabolism ; Diagnosis, Differential ; Female ; Follow-Up Studies ; Gastrointestinal Neoplasms ; Giant Cells ; pathology ; Humans ; Immunohistochemistry ; Immunophenotyping ; Liver Neoplasms ; genetics ; immunology ; metabolism ; pathology ; MART-1 Antigen ; metabolism ; Melanoma-Specific Antigens ; metabolism ; Muscle, Smooth ; metabolism ; Prognosis

OBJECTIVETo study the clinicopathologic features, immunophenotype and genetic changes of perivascular epithelioid cell neoplasms (PEComa).

METHODSA total of 25 cases of PEComa located in various anatomic sites were selected for immunohistochemical staining (SP or EnVision method). TFE3 fluorescence in-situ hybridization was also performed to determine the TFE3 gene status.

RESULTSThe age of patient ranged from 21 to 61 years (mean = 43 years). The male-to-female ratio was 1: 1.3. Histologically, 22 cases represented conventional angiomyolipomas, composed of a mixture of adipose tissue, spindle element, epithelioid smooth muscle cells and abnormal thick-walled blood vessels in various proportions. Three cases involving lung, soft tissue and broad ligament had subtle but distinctive morphologic features. Nested or sheet-like architecture with epithelioid or spindle cells was observed. Immunohistochemical study showed that HMB 45, melan A, smooth muscle actin and cathepsin K were expressed in 80% (20/25), 88% (22/25), 88% (22/25) and 100% (25/25) of PEComa, respectively. Within positive cases, the average proportion of positive tumor cells was 36%, 41%, 35% and 90% respectively for HMB 45, melan A, smooth muscle actin and cathepsin K. TFE3 was negative in all of the 22 renal and hepatic PEComa studied, while it was positive in the 3 cases of extra-hepatorenal PEComa. None of the 25 cases exhibited evidence of TFE3 gene fusion or amplification.

CONCLUSIONSExtra-hepatorenal PEComa have distinctive morphologic features and are associated with TFE3 overexpression. Cathepsin K immunostaining demonstrates high sensitivity and specificity in PEComa, better than other commonly employed immunomarkers. This marker is thus useful in diagnosis of PEComa and distinction with other neoplasms.

Actins ; metabolism ; Adult ; Angiomyolipoma ; metabolism ; pathology ; Basic Helix-Loop-Helix Leucine Zipper Transcription Factors ; genetics ; metabolism ; Cathepsin K ; metabolism ; Female ; Humans ; Immunohistochemistry ; Kidney Neoplasms ; metabolism ; pathology ; Liver Neoplasms ; metabolism ; pathology ; MART-1 Antigen ; metabolism ; Male ; Melanoma-Specific Antigens ; metabolism ; Middle Aged ; Perivascular Epithelioid Cell Neoplasms ; metabolism ; pathology ; Young Adult

Adenoma, Acidophil ; metabolism ; Adult ; Angiomyolipoma ; genetics ; metabolism ; pathology ; Carcinoma, Renal Cell ; metabolism ; Codon ; Diagnosis, Differential ; Exons ; Female ; Follow-Up Studies ; Gene Deletion ; Genes, p53 ; Humans ; Kidney Neoplasms ; genetics ; metabolism ; pathology ; Male ; Melanoma-Specific Antigens ; metabolism ; Middle Aged ; Retrospective Studies ; Tumor Suppressor Protein p53 ; genetics ; metabolism

OBJECTIVETo prepare IL-2-anchored and tumor-derived exosomes vaccine, and investigate the antitumor efficiency of the special cytotoxic T-lymphocytes induced by Ex/GPI-IL-2.

METHODSTo construct pEGFP-N1-IL2gpi plasmid coding a fusion gene of a DNA oligo encoding GPI-anchor signal sequence attaching to human IL-2 cDNA. Then T24 cell lines stably expressing GPI-IL-2 proteins (T24/GPI-IL-2) were established. Ex/GPI-IL-2 were isolated and purified by ultrafiltration and sucrose gradient centrifugation, and the morphology and molecule markers were analyzed. The mixed lymphocyte reaction study and cytotoxic study were performed to determine the proliferative effect of T lymphocytes and the cytotoxicity induced by Ex/GPI-IL-2.

RESULTSThe pEGFP-N1-IL2gpi plasmid was successfully constructed, and cell lines stably expressing GPI-IL-2 fusion proteins were established. Ex/GPI-IL-2 were small vesicular and saucer-shaped in diameter of 30-90 nm, containing heat shock protein 70, intercellular adhesion molecule-1, MAGE-1 and GPI-IL-2. Ex/GPI-IL-2-pulsed could dendritic cells induce proliferation of T cells and cytotoxic immune response more efficiently (P<0.05).

CONCLUSIONSGPI-IL-2 gene-modified tumor cells can make the exosomes containing GPI-IL-2 with an increased anti-tumor effect. Our study provides a feasible approach for exosome-based tumor immunotherapy of bladder transitional cell tumors.

Cancer Vaccines ; immunology ; Carcinoma, Transitional Cell ; immunology ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Exosomes ; genetics ; metabolism ; Glycosylphosphatidylinositols ; metabolism ; HSP70 Heat-Shock Proteins ; metabolism ; Humans ; Intercellular Adhesion Molecule-1 ; metabolism ; Interleukin-2 ; metabolism ; Melanoma-Specific Antigens ; metabolism ; Plasmids ; Recombinant Fusion Proteins ; metabolism ; T-Lymphocytes, Cytotoxic ; immunology ; Transfection ; Urinary Bladder Neoplasms ; immunology ; metabolism ; pathology

BACKGROUND: The 3q29 microdeletion syndrome is a genomic disorder characterized by mental retardation, developmental delay, microcephaly, and slight facial dysmorphism. In most cases, the microdeletion spans a 1.6-Mb region between low-copy repeats (LCRs). We identified a novel 4.0- Mb deletion using oligonucleotide array comparative genomic hybridization (array CGH) in monozygotic twin sisters. METHODS: G-banded chromosome analysis was performed in the twins and their parents. Highresolution oligonucleotide array CGH was performed using the human whole genome 244K CGH microarray (Agilent Technologies, USA) followed by validation using FISH, and the obtained results were analyzed using the genome database resources. RESULTS: G-banding revealed that the twins had de novo 46,XX,del(3)(q29) karyotype. Array CGH showed a 4.0-Mb interstitial deletion on 3q29, which contained 39 genes and no breakpoints flanked by LCRs. In addition to the typical characteristics of the 3q29 microdeletion syndrome, the twins had attention deficit-hyperactivity disorder, strabismus, congenital heart defect, and gray hair. Besides the p21-activated protein kinase (PAK2) and discs large homolog 1 (DLG1) genes, which are known to play a critical role in mental retardation, the hairy and enhancer of split 1 (HES1) and antigen p97 (melanoma associated; MFI2) genes might be possible candidate genes associated with strabismus, congenital heart defect, and gray hair. CONCLUSIONS: The novel 4.0-Mb 3q29 microdeletion found in the twins suggested the occurrence of genomic rearrangement mediated by mechanisms other than nonallelic homologous recombination. Molecular genetic and functional studies are required to elucidate the contribution of each gene to a specific phenotype.
Adaptor Proteins, Signal Transducing/genetics ; Adolescent ; Attention Deficit Disorder with Hyperactivity/genetics ; Basic Helix-Loop-Helix Transcription Factors/genetics ; *Chromosome Deletion ; Chromosome Disorders/*genetics ; *Chromosomes, Human, Pair 3 ; Comparative Genomic Hybridization/*methods ; Diseases in Twins/*genetics ; Female ; Homeodomain Proteins/genetics ; Humans ; In Situ Hybridization, Fluorescence ; Melanoma-Specific Antigens/genetics ; Membrane Proteins/genetics ; Oligonucleotide Array Sequence Analysis ; Syndrome ; Twins ; p21-Activated Kinases/genetics

OBJECTIVETo develop a sensitive and specific RT-PCR assay using the mRNA of MAGE-1 and MAGE-3 genes as specific tumor markers for the detection of the tumor cells in the peripheral blood of patients with gastric cancer.

METHODSPeripheral blood was obtained from 40 patients with gastric cancer and from 20 healthy volunteers. The mRNA of MAGE-1 and MAGE-3 genes in the peripheral blood mononuclear cells (PBMC) was detected by RT-PCR. The expressions of MAGE-1 and MAGE-3 mRNA in the tumor tissues of these gastric cancer patients were also detected by RT-PCR. Meanwhile,CEA expression by nested RT-PCR in PBMC of 40 gastric cancer patients was also detected.

RESULTSOf 40 gastric cancer patients, MAGE-1 and MAGE-3 mRNA were positive in 47.5% (19/40) and 25% (10/40) of PBMC respectively, and in 62.5% (25/40) and 30% (12/40) of gastric cancer tissues respectively. As a whole, in the PBMC of 40 gastric cancer patients, 25 (62.5%) samples were found to express at least one type of MAGE mRNA. In the patients whose tumors did not express MAGE-1 and/or MAGE-3 genes, the corresponding MAGE mRNA was also undetected in their PBMC. There was no expression of MAGE-1 or MAGE-3 gene in the PBMC from the 20 healthy donors. The positive rate of MAGE mRNA in PBMC was closely correlated with the tumor stage and lymph node metastasis (P <0.05). Positive rate of CEA gene expression was 32.5% (13/40) in the PBMC of 40 gastric cancer patients, 29 (72.5%)samples were detected to express at least one type of MAGE gene and CEA gene mRNA.

CONCLUSIONSMAGE-1, MAGE-3 and CEA mRNA are specifically detected with high percentage in the PBMC of gastric cancer patients by RT-PCR. They could be used as specific tumor markers for the detection of the circulating gastric cancer cells, and the detection results may be helpful to evaluate the prognosis of gastric cancer patients.

Adult ; Aged ; Antigens, Neoplasm ; blood ; genetics ; Biomarkers, Tumor ; blood ; Carcinoembryonic Antigen ; blood ; Female ; Humans ; Male ; Melanoma-Specific Antigens ; Middle Aged ; Neoplasm Proteins ; blood ; genetics ; RNA, Messenger ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Sensitivity and Specificity ; Stomach Neoplasms ; blood ; genetics ; pathology

OBJECTIVETo explore the possibility of using melanoma antigen (MAGE)-1 and MAGE-3 gene encoding proteins as an index of potential target for immunotherapy in intrahepatic cholangiocarcinoma (IHCC) patients.

METHODSThe expressions of MAGE-1 and MAGE-3 genes in tumor tissues and tumor adjacent non-IHCC liver tissues were examined by RT-PCR method. The relationship between positive expression rates of MAGE-1 and MAGE-3 genes and clinical data including sex, age, tumor diameters, tumor envelope, tumor nodules number, and hepatitis B virus surface antigen were determined.

RESULTSThe positive expression rates of MAGE-1 (35%) and MAGE-3 genes (45%) were significantly higher in the tumor tissues than in tumor adjacent tissues (0) (P<0.01). The positive expression rates of MAGE-1 and MAGE-3 genes had no relationship with the clinical data (P >0.05), except the morphology of tumor (P <0.05).

CONCLUSIONThe high expression rates of MAGE-1 and MAGE-3 genes in IHCC suggests the MAGE-1 and MAGE-3 gene may be a target for immunotherapy in IHCC patients.

Adult ; Aged ; Antigens, Neoplasm ; genetics ; Bile Duct Neoplasms ; genetics ; Bile Ducts, Intrahepatic ; pathology ; Cholangiocarcinoma ; genetics ; Female ; Humans ; In Vitro Techniques ; Liver Neoplasms ; genetics ; Male ; Melanoma-Specific Antigens ; Middle Aged ; Neoplasm Proteins ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo provide an useful animal model for exploring metastatic biology and anti-metastatic therapy of primary malignant melanoma of the small intestine.

METHODSA 49-year old male patient with malignant melanoma was treated by surgery, and the primary tumor in the small intestine and a metastatic tumor in the liver were removed. The diagnosis of malignant melanoma was confirmed by histopathology. Fresh melanoma tissue fragments taken from the primary intestinal tumor and hepatic metastatic tumor were orthotopically implanted into the mucosal layer of small intestine in nude mice, respectively. The tumor growth rate, invasion and metastasis of the transplanted tumors were observed. Light and electron microscopy, immunophenotype analysis, flow cytometry and karyotype analysis were carried out.

RESULTSFragments of the primary and liver metastatic malignant melanoma were successfully implanted in nude mice. After continuous passages in nude mice, an highly-metastatic model of human primary malignant melanoma of the small intestine (from the primary lesion) in nude mice (termed HSIM-0602) and a liver metastatic model of human primary malignant melanoma of the small intestine (originally from the liver metastatic lesion) in nude mice (termed HSIM-0603) were successfully established. Histological examination of the transplanted tumors revealed a high-grade melanoma of the small intestine. Immunohistochemical stainings of S-100 protein and HMB45 were positive. Many scattered melanosomes and melanin complex were seen in the cytoplasm of tumor cells. Chromosomal modal number was between 55 and 59. DNA index (DI) was 1.59 - 1.71, representing a heteroploid. The HSIM-0602 and HSIM-0603 tumor models had been maintained for 21 and 23 passages in nude mice, respectively. 227 nude mice were used for transplantation. Both the growth rate after transplantation and resuscitation rate from liquid nitrogen cryopreservation were 100%. The HSIM-0602 model exhibited 84.8% lung metastasis, 65.7% liver metastasis and 63.8% lymph node metastasis. However, HSIM-0603 displayed 100% liver metastasis, 46.7% lung metastasis and 71.3% lymph node metastasis. The transplanted tumors actively and invasively grew in the small intestine of nude mice and showed hematogenous and lymphatic metastases.

CONCLUSIONTo our knowledge it is the first time that two strains of spontaneous highly-metastatic nude-mouse model of human primary malignant melanoma of the small intestine have been successfully established in our department. The models are very closely mimic the natural clinicopathologic course of primary small intestinal melanoma in humans and provide ideal animal models for the researches on metastasis biology and anti-metastatic experimental therapy of malignant melanoma of the small intestine.

Animals ; Antigens, Neoplasm ; metabolism ; DNA, Neoplasm ; genetics ; Disease Models, Animal ; Female ; Humans ; Intestine, Small ; Jejunal Neoplasms ; genetics ; pathology ; secondary ; ultrastructure ; Liver Neoplasms ; genetics ; pathology ; secondary ; Lung Neoplasms ; genetics ; pathology ; secondary ; Lymphatic Metastasis ; Male ; Melanoma ; genetics ; pathology ; ultrastructure ; Melanoma-Specific Antigens ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Microscopy, Electron ; Middle Aged ; Neoplasm Proteins ; metabolism ; Neoplasm Transplantation ; Polyploidy ; S100 Proteins ; metabolism

OBJECTIVESTo investigate the correlation between MAGE-A1 mRNA expression and genic demethylation in hepatoma cell lines.

METHODSTotal RNA and genomic DNA were prepared from 10 human hepatoma cell lines. MAGE-1 mRNA expression was determined with RT-PCR and the level of genome-wide demethylation was evaluated by enzyme digestion and Southern blot assay. The genomic DNA was digested by HpaII, then the promoter of MAGE-A1 gene was amplified with primers CDS21, EDP4 and CDS20. EDP4 and the PCR products were further hybridized with a probe to detect the methylation in the promoter of the MAGE-A1 gene. HLA-A locus was typed using SSP kit.

RESULTSIn cell lines QGY-7703, SMMC-7721, HLE, BEL-7402, BEL-7404 and BEL-7405, MAGE-A1 mRNA expression was positive and cell differentiation was moderate or low. In cell lines of HepG2 2.2.15, HepG2, QGY-7701 and Huh7, MAGE-A1 mRNA expression was negative and cell differentiation was well or moderate. The level of genomic demethylation in MAGE-A1 mRNA positive cell lines was much higher than that in MAGE-A1 mRNA negative cell lines (t = 2.896, P = 0.02). The methylation analysis showed that methylation in the promoters of MAGE-A1 gene of HepG2 2.2.15, HepG2, QGY-7701 and Huh7 was high, and that methylation in those of SMMC-7721, HLE, BEL-7402, BEL-7404, and BEL-7405 was low.

CONCLUSIONThe results suggest that MAGE-A1 mRNA expression in the human hepatoma cell lines is associated with genic hypomethylation.

Antigens, Neoplasm ; Carcinoma, Hepatocellular ; genetics ; immunology ; metabolism ; DNA Methylation ; Humans ; Liver Neoplasms ; genetics ; immunology ; metabolism ; Melanoma-Specific Antigens ; Neoplasm Proteins ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Tumor Cells, Cultured

OBJECTIVETo evaluate the significance of melanoma antigen (MAGE) gene expression in bladder transitional cell carcinoma (TCC).

METHODSMAGE-A1, A2, A3, A4 mRNA expression was detected by reverse transcription polymerase chain reaction (RT-PCR) in 3 clusters bladder TCC cells and 20 samples of bladder TCC patients (T(1) 7 samples, T(2) 5 samples, T(3) 6 samples, T(4) 2 samples, G(1) 1 samples, G(2) 11 samples, G(3) 8 samples). MAGE-A4 protein was detected by immunohistochemistry in 105 samples of bladder TCC patients (T(1) 35 samples, T(2) 12 samples, T(3) 26 samples, T(4) 13 samples, G(1) 13 samples, G(2) 44 samples, G(3) 48 samples).

RESULTSThree clusters bladder TCC cells had MAGE gene mRNA expression. In detection of MAGE mRNA of 20 samples of bladder TCC patients, 12 samples (60%) expressed MAGE-A1, 16 samples (80%) expressed MAGE-A2, 11 samples (55%) expressed MAGE-A3, 18 samples (90%) expressed MAGE-A4, 8 samples (40%) expressed all of MAGE-A1--A4. In 105 bladder TCC samples, 53 samples (50%) expressed MAGE-A4 protein. Strong expression (++ or +++) was significant higher in higher grade (13 samples/48 samples) or stage (14 samples/51 samples) than in lower grade (2 samples/57 samples) or stage (0 samples/35 samples).

CONCLUSIONMAGE gene highly expresses in bladder TCC. Bladder TCC of high grade or high stage has higher MAGE-A4 protein strong expression.

Adult ; Aged ; Aged, 80 and over ; Antigens, Neoplasm ; biosynthesis ; genetics ; Carcinoma, Transitional Cell ; genetics ; metabolism ; Female ; Gene Expression ; Humans ; Male ; Melanoma-Specific Antigens ; Middle Aged ; Neoplasm Proteins ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; Reverse Transcriptase Polymerase Chain Reaction ; Tumor Cells, Cultured ; Urinary Bladder Neoplasms ; genetics ; metabolism