PURPOSE: Although breast cancer the most common cancer for women remains a significant health problem, it has not been systematically studied until now. In an attempt to identify novel genes implicated in breast cancer development, we performed a suppression subtraction hybridization (SSH) with human breast cancer tissues, as well as with cloned genes, that are expressed more than in normal tissue. METHODS: After the identification of a novel gene, RT-PCR was performed to determine its mRNA expression in human breast cancers. In order to learn more about the expression profile of this gene, PCR was performed using various commercially available normal or carcinoma cell lines. The novel gene was found to be strongly expressed in breast cancer tissues and other carcinoma cell lines. To determine whether this novel gene was associated with cell cycle regulation, normal WI-38 fibroblast cells were stimulated with media containing 0.1% FBS for 48hours. RESULT: From the experimental results of the SSH, a novel clone, "Clone 135" which was strongly expressed in tumor compared to matched normal tissue, has been found. The novel clone was identified as being expressed in several tumor tissues and carcinoma cell lines. The time-course expression of this novel gene in the WI-38 (8PDL) normal lung cell line indicated a significant increase for G1-phase arrest. CONCLUSION: We have used a suppression subtractive hybridization (SSH) to generate a profile of genes overexpressed in human breast cancer. We have screened novel genes, of which "Clone 135" scored as a candidate oncogene that was overexpressed in tumor compared to matched normal tissue.
Breast Neoplasms* ; Breast* ; Cell Cycle ; Cell Line ; Clone Cells ; Female ; Fibroblasts ; Humans* ; Lung ; Oncogenes ; Polymerase Chain Reaction ; RNA, Messenger

PURPOSE: Although breast cancer the most common cancer for women remains a significant health problem, it has not been systematically studied until now. In an attempt to identify novel genes implicated in breast cancer development, we performed a suppression subtraction hybridization (SSH) with human breast cancer tissues, as well as with cloned genes, that are expressed more than in normal tissue. METHODS: After the identification of a novel gene, RT-PCR was performed to determine its mRNA expression in human breast cancers. In order to learn more about the expression profile of this gene, PCR was performed using various commercially available normal or carcinoma cell lines. The novel gene was found to be strongly expressed in breast cancer tissues and other carcinoma cell lines. To determine whether this novel gene was associated with cell cycle regulation, normal WI-38 fibroblast cells were stimulated with media containing 0.1% FBS for 48hours. RESULT: From the experimental results of the SSH, a novel clone, "Clone 135" which was strongly expressed in tumor compared to matched normal tissue, has been found. The novel clone was identified as being expressed in several tumor tissues and carcinoma cell lines. The time-course expression of this novel gene in the WI-38 (8PDL) normal lung cell line indicated a significant increase for G1-phase arrest. CONCLUSION: We have used a suppression subtractive hybridization (SSH) to generate a profile of genes overexpressed in human breast cancer. We have screened novel genes, of which "Clone 135" scored as a candidate oncogene that was overexpressed in tumor compared to matched normal tissue.
Breast Neoplasms* ; Breast* ; Cell Cycle ; Cell Line ; Clone Cells ; Female ; Fibroblasts ; Humans* ; Lung ; Oncogenes ; Polymerase Chain Reaction ; RNA, Messenger

OBJECTIVE: A variety of genetic alterations in human glioblastoma comprises signal transduction and cell cycle arrest control of cellular processes. Subtractive hybridization is potentially a faster method for identifying differentially expressed genes associated with a particular disease state. Using the technique of subtraction, we isolated novel genes that are overexpressed in glioblastoma tissue as compared to normal brain tissue. METHODS: We evaluated the differential expression of genes in each of hybridizing tester and driver cDNAs to digested 130 clones. After sequencing of 130 clones and homology search, this study performed to determine mRNA expression of the unknown gene, "clone 47", in brain tissue, glioblasoma, and several cancer cell lines by reverse transcription-polymerase chain reaction (RT-PCR). To test the time course for G0-phase arrest, serum stimulation and expression at various times for RT-PCR performed. RESULTS: We identified 23 novel genes by BLAST of the digested 130 clones. The expressions of "clone 47" mRNA of glioblastoma and several cancer lines were significantly higher than normal brain tissues and several normal cell lines. We confirmed the mRNA expression of "clone 47" was up-regulation for 0.5~1hr of WI-38 cell differentiation. CONCLUSION: The novel gene, "Clone 47" is upregulated in glioblastoma tissue and several cancer cell lines. This gene is time dependent activation during time course of serum stimulation. This result suggests that "clone 47" play a role in brain tumorigenesis and the activation of this "clone 47" may be necessary for the development of cancer.
Brain ; Carcinogenesis ; Cell Cycle Checkpoints ; Cell Differentiation ; Cell Line ; Clone Cells ; DNA, Complementary ; Glioblastoma* ; Humans ; RNA, Messenger ; Signal Transduction ; Up-Regulation

PURPOSE: This study was done to review our clinical experience of fluorodeoxyglucose (FDG)-positron emission tomography (PET) in biliary-pancreatic malignancies, and evaluated its diagnostic accuracy and usefulness. METHODS: FDG-PET was performed in 15 patients with biliary-pancreatic malignancies, to determine the extent of the disease (n=9) and to detect the recurrence (n=6). RESULTS: Of the 9 patients with a suspected loco-regional spread, 6 patients were disclosed by FDG-PET as having multiple metastasis to the regional lymph nodes and extraperitoneal organs, but the other 3 showed no evidence of regional or distant metastasis. Thus, FDG-PET performed preoperatively to estimate the extent of the disease, showed sensitivity and specificity of 100%, respectively. However, for the detection of a recurrent disease only 3 out of 5 with recurrent diseases were detected by FDG-PET (sensitivity: 60%). FDG-PET revealed one suspected recurrent lesion, but a computed tomography (CT) scan showed no evidence of a recurrence. Due to the 2 false-negative results, the overall sensitivity and specificity of this study were 81.8 (9/11) and 100% (4/4), respectively. CONCLUSION: Unnecessary surgical procedures were avoided in 6 cases, and 3 curative surgeries aborted through the use of FDG-PET. Despite the lack of cases, FDG-PET was useful in estimating the extent of a disease, but was unsatisfactory in detecting a recurrent disease compared to CT. Coupled with conventional imaging, FDG-PET may also provide more helpful information for biliary-pancreatic malignancies as well.
Humans ; Lymph Nodes ; Neoplasm Metastasis ; Recurrence ; Sensitivity and Specificity

PURPOSE: Second-generation tyrosine kinase inhibitors (second TKIs) such as nilotinib and dasatinib control the activity of most ABL kinase domain mutations observed in patients with imatinib resistance. Although in vitro data show that both agents can inhibit all mutations except T315I, some discrepancies have been observed in a small subset of mutation clones. Cytogenetic clonal evolution is the important resistance mechanism of chronic myeloid leukemia (CML). Accordingly, we observed the clinical significance of coexisting with clonal evolution and BCR-ABL mutant in CML patients treated with second TKIs. MATERIALS AND METHODS: We monitored BCR-ABL transcript kinetics, interrelationship of clones expressing non-mutated and mutant transcripts and clonal aberrations within Philadelphia (Ph) positive and negative clones, respectively, in eight patients with CML receiving dasatinib or nilotinib for 3~41 months. RESULTS: Clinical responses were correlated with in vitro sensitivity of the BCR-ABL mutants to the second TKIs in four patients. Four patients showed resistance to the second TKIs as compared to in vitro observations; three of them developed chromosomal abnormalities in the Ph chromosome positive or negative metaphases. Another patient lost the original mutation but acquired a more resistant new mutation and became resistant to the second TKI. CONCLUSION: Cytogenetic clonal evolution is an independent poor prognostic factor in CML, which could explain the onset of mechanisms for second TKIs resistance to ABL kinase domain mutations. The results indicate that an additional evaluation of chromosomal abnormalities is warranted when BCR-ABL mutants are more resistant than indicated by in vitro data.
Benzamides ; Chromosome Aberrations ; Clonal Evolution ; Clone Cells ; Cytogenetics ; Dasatinib ; Humans ; Hydrogen-Ion Concentration ; Kinetics ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; Metaphase ; Philadelphia ; Phosphotransferases ; Piperazines ; Protein-Tyrosine Kinases ; Pyrimidines ; Thiazoles ; Tyrosine ; Imatinib Mesylate