Good progress has been made in the researches on correlating genes of breast cancer in recent years. Quite a few kinds of genes such as susceptibility gene, oncogene and tumor suppressor genes have been found with implications for diagnosis, therapy and prognosis. Abnormality of breast cancer susceptibility gene (BRCA) is of great significance, especially in the development of breast cancer.
BRCA1 Protein ; genetics ; BRCA2 Protein ; genetics ; Breast Neoplasms ; genetics ; Cyclin-Dependent Kinase Inhibitor p16 ; genetics ; Cyclin-Dependent Kinase Inhibitor p21 ; Cyclins ; genetics ; Female ; Humans ; Mutation ; Neoplasm Proteins ; genetics ; Proto-Oncogene Proteins ; genetics ; Tumor Suppressor Protein p53 ; genetics

The aim of this study was to investigate the nucleotide sequence of one distinct fusion transcript of sil-tal1 in childhood T-ALL. The PCR product was cloned into plasmid vector and then sequenced. Genomic DNA was analyzed with PCR using the designed primer pairs representing distinct sequences. The product was sequenced and analyzed with database. The results indicated that 4 different fusion transcripts were detected at cDNA level, in which a part of exons or introns of sil are reserved respectively, and some additions and deletions existed. After analyzing genomic DNA sequence of leukemic cells, the breakpoint in gene sil of this case was proved to be different at DNA level from references. Hence, the sil-tal1 rearrangement was defined to be a new type. It is concluded tal1 rearrangement of leukemic cells in this case is a new type, which expresses classical and at least 3 variant fusion transcripts, presumably caused by extraordinary mechanisms of splicing and transcription in leukemic stem cells.
Base Sequence ; Basic Helix-Loop-Helix Transcription Factors ; genetics ; Child ; DNA Breaks ; DNA, Neoplasm ; genetics ; Humans ; Leukemia-Lymphoma, Adult T-Cell ; genetics ; Male ; Oncogene Proteins, Fusion ; genetics ; Proto-Oncogene Proteins ; genetics ; T-Cell Acute Lymphocytic Leukemia Protein 1 ; Transcription Factors

OBJECTIVETo study the effects of anti-ABL tyrosine kinase intrabody on the growth of human chronic myelogenous leukemia (CML) cells in nude mice.

METHODSA recombinant retroviral vector MSCV-ibE-IRES-eGFP was constructed to express intracellular single-chain antibody (intrabody) against ABL tyrosine kinase domain in CML cells. K562 cells were transduced with the retrovirus, eGFP+ cells were then selected by fluorescence-activated cell sorting (FACS). The intrabody mRNA expression was determined by reverse transcription (RT)-polymerase chain reaction (PCR). BCR/ABL and c-ABL protein tyrosine kinase (PTK) activity in the cells was examined. Transduced cells and control group K562 cells were transplanted into nude mice respectively and the tumor sizes were dynamically observed.

RESULTSK562-ibE cell was obtained. Expression of the BCR/ABL and c-ABL protein tyrosine kinase activity of harvested K562-ibE cells were markedly inhibited. At 14, 21 and 28 days after cell injection, the tumor volumes of experimental mice were obviously smaller than that of control mice, about one half of the control groups (P < 0.05).

CONCLUSIONThe growth of K562-ibE cells was significantly inhibited in vivo. It is possible that inhibition of the BCR/ABL protein tyrosine kinase activity by the intrabody blocked BCR/ABL signal transduction pathway, promoted apoptosis and reduced tumorigenicity of K562 cells in vivo.

Animals ; Antibodies ; genetics ; Apoptosis ; Cell Division ; Fusion Proteins, bcr-abl ; biosynthesis ; genetics ; immunology ; Genetic Vectors ; Humans ; K562 Cells ; Mice ; Mice, Nude ; Neoplasm Transplantation ; Protein-Tyrosine Kinases ; immunology ; metabolism ; Proto-Oncogene Proteins c-abl ; genetics ; immunology ; Retroviridae ; genetics

OBJECTIVE: To investigate the effects of recombinant human thrombopoietin (rhTPO) to proliferation and apoptosis of acute myeloid leukemia (AML) cell lines. METHODS: After the treatment of different concentrations of rhTPO (0, 50, 100 ng/ml) for different time (24,48,72 h)，the cell proliferation rates of the AML cell lines (Kasumi-1, Skno-1, HEL, HL-60, THP-1) were determined by CCK-8 method. Apoptosis rate of each cell line cocultured with rhTPO was detected by Annexin V/PI method. The relative expression of TPO receptor c-MPL (myeloproliferative clonal antibody) mRNA in AML cell lines was detected by Q-PCR. The expression of c-MPL protein in each cell line was detected by Western blot. The expression of c-MPL antigen in HL-60 cells treated by different concentrations of rhTPO was detected by Flow cytometry. RESULTS: RhTPO showed no promotion to the proliferation of Kasumi-1, Skno-1, HEL, HL-60, THP-1 cell lines，however，it showed inhibitory effect to cell proliferation (72 h 0 ng/ml vs 100 ng/ml, P= 0.029) and pro-apoptotic (48 h 0 ng/ml vs 50 ng/ml, P=0.0143) in HL-60 cells. In Kasumi-1, Skno-1, HEL and THP-1 cells, there showed no statistically significant differences in apoptosis rate among each groups treated by different concentrations of rhTPO. Each AML cell line showed different levels of c-MPL gene and c-MPL protein expression, but HEL cells showed the highest expression in both of them. After HL-60 cells were treated by different concentrations of rhTPO for 48 hours, there showed no statistical difference in c-MPL antigen expression among each groups. CONCLUSION RhTPO can not promote the proliferation of Kasumi-1, Skno-1, HEL, HL-60 and THP-1 leukemia cell lines. On the contrary, rhTPO can inhibit HL-60 cell proliferation and promote its apoptosis, and this effect is not related to c-MPL gene expression or protein expression.
Apoptosis ; Cell Proliferation ; Humans ; Leukemia, Myeloid, Acute ; Neoplasm Proteins ; Proto-Oncogene Proteins ; Receptors, Cytokine ; Thrombopoietin

Resistance to thyroid hormone (RTH) is an autosomal dominant disorder that's characterized by inappropriate normal or elevated TSH levels despite of the elevated thyroid hormone levels. RTH is distinguished from the TSH secreting pituitary adenoma by performing the TRH stimulation test, TSH alpha subunit measurement and sellar MRI. A 23 year old woman visited our hospital complaining of fatigue, palpitation and heat intolerance and she had an anterior neck mass. She had elevated total T3, free T4 and TSH levels. The serum TSH levels were increased during the TRH stimulation test before and after T3 suppression. The serum TSH alpha subunit showed a normal response and the serum TSH alpha subunit/TSH molar ratio did not increase over 1.0 with TRH stimulation. Thyroid hormone receptor beta gene mutation was identified. Although a left pituitary microadenoma was revealed on sellar MRI, the patient was diagnosed as having pituitary RTH with a nonfunctioning pituitary microadenoma. We report here on a patient with pituitary RTH and a nonfunctioning pituitary microadenoma, and this is the first such case in Korea.
Fatigue ; Female ; Glycoprotein Hormones, alpha Subunit ; Hot Temperature ; Humans ; Molar ; Neck ; Pituitary Neoplasms ; Thyroid Gland ; Thyroid Hormone Receptors beta

OBJECTIVE: To determine the expression of apoptosis related gene PDCD5 in multiple myeloma (MM), and to analyze the relation between PDCD5 and BCL-2. METHODS: The expressions of PDCD5 and BCL-2 protein and mRNA were determined by immunohistochemical staining method, flow cytometry (FCM) and reverse transcription polymerase chain reaction (RT-PCR) method in bone marrow mononuclear cells. We also analyzed the relation between PDCD5 and BCL-2. RESULTS: Immunohistochemical staining showed that PDCD5 protein positive cell percentage, staining intensity index (SII) of PDCD5 protein, BCL-2 protein positive cell percentage, and SII of BCL-2 protein were (34.75 +/- 6.49)%, (281.16 +/- 75.33), (29.97 +/- 5.57)%, and (224.94 +/- 57.72) in the MM group and (52.98 +/- 5.84)%, (462.84 +/- 39.77), (5.56 +/- 1.95)%, and (27.84 +/- 9.75) in the control group (all P < 0.05). Results of FCM showed that PDCD5 protein positive percentage and mean fluorescence intensity of PDCD5 were (78.11 +/- 21.63)% and (61.73 +/- 11.04) in the MM group and (89.46 +/- 9.98)% and (353.04 +/- 123.26) in the control group (all P < 0.05). RT-PCR showed that relative expression of PDCD5 and BCL-2 mRNA were (0.33 +/ -0.07) and (0.33 +/- 0.08) in the MM group and (0.53 +/- 0.05) and (0.12 +/- 0.02) in the control group (all P < 0.05). The positive cell percentage of PDCD5 and BCL-2 protein was negative correlation (r = -0.86, P < 0.05); the expression of PDCD5 and BCL-2 mRNA was the same status (r = -0.90, P < 0.05). CONCLUSION The expressions of PDCD5 protein and mRNA in MM patients are down-regulated, but the expressions of BCL-2 protein and mRNA are up-regulated. The mRNA and protein expression of PDCD5 and BCL-2 has negative correlation.
Adult ; Aged ; Apoptosis ; genetics ; Apoptosis Regulatory Proteins ; biosynthesis ; genetics ; Bone Marrow Cells ; metabolism ; pathology ; Female ; Humans ; Male ; Middle Aged ; Multiple Myeloma ; genetics ; metabolism ; pathology ; Neoplasm Proteins ; biosynthesis ; genetics ; Proto-Oncogene Proteins c-bcl-2 ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics

PURPOSE: Dermatofibrosarcoma protuberans (DFSP) carries a translocation resulting in the collagen type I alpha 1 (COL1A1)-platelet-derived growth factor beta (PDGFB) fusion gene, which is responsible for PDGFB activation. The purpose of this study is to evaluate the clinicopathological, genetic, and therapeutic features of DFSP in Korean patients. MATERIALS AND METHODS: Clinicopathological features of 37 patients with DFSP were reviewed. Multiplex reverse transcriptase-polymerase chain reaction (PCR) was carried out in 16 patients using formalin-fixed, paraffin-embedded tissues and specific primers for COL1A1 and PDGFB. RESULTS: The mean age of 37 patients was 37.4 years old. The most common tumor location was the trunk. All patients were treated primarily with surgery: 34 (91.7%) cases with Mohs micrographic surgery (MMS) and 3 (8.3%) cases with wide local excision. The median follow-up time was 33.7 months. Two patients, one in each treatment group, demonstrated local recurrence during the follow-up period. The COL1A1-PDGFB fusion gene was expressed in 14 (87.5%) cases, demonstrated by reverse transcriptase PCR analysis. No association was found among the different COL1A1-PDGFB fusion transcripts, the various histological subtypes and clinical features. CONCLUSION: Our results support the effectiveness of MMS in treating DFSP. The COL1A1-PDGFB fusion transcript was observed in 87.5% of patients. Therefore, COL1A1-PDGFB is a useful and accurate tool in diagnosing DFSP in Koreans.
Adolescent ; Adult ; Asian Continental Ancestry Group/*genetics ; Collagen Type I/*genetics ; DNA Primers ; Dermatofibrosarcoma/ethnology/*genetics/*pathology/surgery ; Female ; Humans ; Male ; Middle Aged ; Mohs Surgery ; Multiplex Polymerase Chain Reaction ; Neoplasm Recurrence, Local ; Oncogene Proteins, Fusion/*genetics ; Proto-Oncogene Proteins c-sis/*genetics ; Republic of Korea ; Reverse Transcriptase Polymerase Chain Reaction ; Skin Neoplasms/ethnology/*genetics/*pathology/surgery ; Treatment Outcome

PURPOSE: Dermatofibrosarcoma protuberans (DFSP) carries a translocation resulting in the collagen type I alpha 1 (COL1A1)-platelet-derived growth factor beta (PDGFB) fusion gene, which is responsible for PDGFB activation. The purpose of this study is to evaluate the clinicopathological, genetic, and therapeutic features of DFSP in Korean patients. MATERIALS AND METHODS: Clinicopathological features of 37 patients with DFSP were reviewed. Multiplex reverse transcriptase-polymerase chain reaction (PCR) was carried out in 16 patients using formalin-fixed, paraffin-embedded tissues and specific primers for COL1A1 and PDGFB. RESULTS: The mean age of 37 patients was 37.4 years old. The most common tumor location was the trunk. All patients were treated primarily with surgery: 34 (91.7%) cases with Mohs micrographic surgery (MMS) and 3 (8.3%) cases with wide local excision. The median follow-up time was 33.7 months. Two patients, one in each treatment group, demonstrated local recurrence during the follow-up period. The COL1A1-PDGFB fusion gene was expressed in 14 (87.5%) cases, demonstrated by reverse transcriptase PCR analysis. No association was found among the different COL1A1-PDGFB fusion transcripts, the various histological subtypes and clinical features. CONCLUSION: Our results support the effectiveness of MMS in treating DFSP. The COL1A1-PDGFB fusion transcript was observed in 87.5% of patients. Therefore, COL1A1-PDGFB is a useful and accurate tool in diagnosing DFSP in Koreans.
Adolescent ; Adult ; Asian Continental Ancestry Group/*genetics ; Collagen Type I/*genetics ; DNA Primers ; Dermatofibrosarcoma/ethnology/*genetics/*pathology/surgery ; Female ; Humans ; Male ; Middle Aged ; Mohs Surgery ; Multiplex Polymerase Chain Reaction ; Neoplasm Recurrence, Local ; Oncogene Proteins, Fusion/*genetics ; Proto-Oncogene Proteins c-sis/*genetics ; Republic of Korea ; Reverse Transcriptase Polymerase Chain Reaction ; Skin Neoplasms/ethnology/*genetics/*pathology/surgery ; Treatment Outcome

BACKGROUND AND OBJECTIVEPrimary renal synovial sarcoma is rare and might be misdiagnosed as another renal tumor. This study demonstrates the clinicopathologic and immunohistochemical features, differential diagnosis, and prognosis of such tumors.

METHODSHistologic slides and clinical data were reviewed for 4 patients with primary renal synovial sarcoma and immunohistochemical staining was performed. Molecular analysis was performed on 2 cases to demonstrate the presence of the SYT-SSX gene fusion transcripts by reverse transcriptase polymerase chain reaction (RT-PCR).

RESULTSThe patients were 2 women and 2 men aged from 32 to 48 years. The tumors were 10.0-15.0 cm in diameter, grey-white and solid, and hemorrhage or necrosis was observed. Microscopically, the tumors consisted of mitotically active, monomorphic plump spindle cells with indistinct cell borders growing in short, intersecting fascicles. Hypocellular myxoid areas and a prominent hemangiopericytomatous pattern were present in all cases. The average mitotic rate was 5-8 mitoses/10 high-power fields. Hemorrhage and tumor necrosis were easily found. Scattered small cysts lined with flat, cuboidal, or hobnailed epithelia were found in 3 cases. Tumor cells are immunoreactive for Vimentin (4/4), Bcl-2 (4/4), CD99 (4/4), and CD56 (3/4), and focally for EMA (3/4) and Cytokeratin (3/4). SYT-SSX1 gene fusion was detected in the 2 cases in which RT-PCR analysis was performed. One patient had tumor metastasis to the lung 6 months after surgery and died 5 months later. Multiple metastasis to the liver occurred in one patient and the patient died 13 months after the initial surgery. The other 2 patients had tumors recur at 8 and 15 months and died at 18 and 21 months, respectively, after the initial operation.

CONCLUSIONPrimary renal synovial sarcoma is rare, with poor prognosis, characterized by SYT-SSX gene fusion, and needs to be differentiated from other renal sarcomas.

12E7 Antigen ; Adult ; Antigens, CD ; metabolism ; CD56 Antigen ; metabolism ; Cell Adhesion Molecules ; metabolism ; Female ; Follow-Up Studies ; Humans ; Keratins ; metabolism ; Kidney Neoplasms ; metabolism ; pathology ; surgery ; Liver Neoplasms ; secondary ; Lung Neoplasms ; secondary ; Male ; Middle Aged ; Mucin-1 ; metabolism ; Neoplasm Recurrence, Local ; Nephrectomy ; Oncogene Proteins, Fusion ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Sarcoma, Synovial ; metabolism ; pathology ; secondary ; surgery ; Survival Rate ; Vimentin ; metabolism

OBJECTIVETo investigate the relationship between topoisomerase II inhibitors and t(8;21) chromosomal translocation.

METHODSThe rearrangements of AML1 and ETO genes were detected by Southern Blot and the AML1-ETO fusion gene by nested RT-PCR combined with sequencing in K562 cells treated with etoposide (Vp16) and doxorubicin (DOX).

RESULTSThe rearrangements of AML1 gene were detectable after DOX treatment at concentrations of 10, 50 and 100 micro mol/L for 16 h, AML1-ETO fusion gene appeared after 50 micro mol/L DOX treatment for 48 h.

CONCLUSIONInduction of AML1 gene rearrangement and fusion by topoisomerase II inhibitors, represents one of the molecular mechanisms of t(8;21) chromosomal translocation.

Apoptosis ; drug effects ; Base Sequence ; Blotting, Southern ; Cell Survival ; drug effects ; Core Binding Factor Alpha 2 Subunit ; DNA, Neoplasm ; genetics ; DNA-Binding Proteins ; genetics ; Doxorubicin ; pharmacology ; Etoposide ; pharmacology ; Gene Rearrangement ; drug effects ; HL-60 Cells ; Humans ; K562 Cells ; Molecular Sequence Data ; Oncogene Proteins, Fusion ; genetics ; Proto-Oncogene Proteins ; genetics ; RNA, Neoplasm ; genetics ; RUNX1 Translocation Partner 1 Protein ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Homology, Nucleic Acid ; Topoisomerase II Inhibitors ; Transcription Factors ; genetics