BACKGROUND: Abnormalities of genomic methylation patterns have been shown to play a role in the development of carcinoma, and the silencing of tumor suppressor genes is related to local de novo methylation. METHODS: Using methylation specific arbitrarily primed-Polymerase Chain Reaction (Ms AP-PCR), we identified a 322 bp sequence that contained a 5' un-translated and exon1 regions of the TPEF gene. To evaluate the inactivation of the TPEF gene through hypermethylation in hepatocellular carcinoma (HCC), we investigated the correlation between methylation patterns and TPEF expression in tumor tissues of human HCC and cell lines via a Combined Bisulfite Restriction Assay (CoBRA) and RT-PCR. RESULTS: A dense methylation pattern of the TPEF was detected in most cell lines, as well as in 10 of the 14 (71.4%) HCC tissues. In addition, loss of heterozygosity (LOH) from the TPEF gene was observed in 5 of the 14 (36%) HCC tissues. Furthermore, RT-PCR analysis revealed TPEF expression in 5 of 8 (62.5%) cell lines. Finally, treatment with a demethylating agent, 5-Aza- 2'-deoxycitidine (5-AzaC), increased the expression of TPEF mRNA. CONCLUSION: These results indicate that inactivation of the TPEF gene through hypermethylation may be a mechanism by which tumorigenesis occurs in HCC.
Humans ; Carcinoma, Hepatocellular ; Cell Transformation, Neoplastic ; Genes, Tumor Suppressor

PURPOSE: Irinotecan, in combination with leucovorin/ 5-fluorouracil (FU) or with cisplatin, is known to be active for treating advanced gastric cancer (AGC). This pilot study evaluated a novel three-drug combination of irinotecan, leucovorin/FU and cisplatin as a first-line treatment of AGC. The primary endpoint was to assess the feasibility in anticipation of conducting a larger phase II study. MATERIALS AND METHODS: Chemotherapy-naive AGC patients received irinotecan 150 mg/m2 on day 1, and leucovorin 200 mg/m2 and a 22-h infusion of FU 1000 mg/m2 on days 1 and 2. Cisplatin 30 mg/m2 was administered on day 2. Treatment was repeated every 2 weeks until disease progression or unacceptable toxicity. RESULTS: Of the 17 eligible patients, two patients had an ECOG performance status of 2 and their median age was 48 years (range: 31 to 69). A total of 117 chemotherapy cycles were delivered (median: 6, range: 1 to 12). The causes of treatment discontinuation were disease progression in 9 patients (53%), refusal (35%) and toxicity (12%). Although grade 3 or 4 neutropenia (41% of patients) was the major toxicity that required dose adjustments, only one episode of febrile neutropenia occurred. Grade 3 or 4 nausea and vomiting, diarrhea and fatigue were observed in 35%, 35% and 29% of patients, respectively. None of the patients died of toxicity during treatment. Of the 16 patients who were evaluable for response, 7 (44%) experienced a partial response. CONCLUSION: This novel multi-drug combination was tolerated well in patients with AGC. Based on the encouraging efficacy and tolerability, a randomized phase II study is ongoing in this disease setting.
Cisplatin* ; Diarrhea ; Disease Progression ; Disulfiram ; Drug Therapy ; Drug Therapy, Combination* ; Fatigue ; Febrile Neutropenia ; Fluorouracil* ; Humans ; Leucovorin* ; Nausea ; Neutropenia ; Pilot Projects* ; Stomach Neoplasms* ; Vomiting

PURPOSE: This report introduces a new method for tensioning the remnant PCL with a reconstruction of the anterolateral (AL) bundle of the PCL using a modified tibial inlay technique with an assessment of the outcome of this method in chronic PCL injury. MATERIALS AND METHODS: From January 1998 to August 2003, eighty six patients was underwent tensioning of a laxed remnant PCL with a reconstruction of the anterolateral bundle of the PCL. Of these, fifty two patients who were followed up for more than 2 years were evaluated. Tensioning was performed using a distal transfer of the tibial attachment with the posteromedial approach in the supine position. The AL bundle of the PCL was reconstructed with 4 bundles of a hamstring autograft or tibialis anterior tendon allograft. The stability was assessed objectively using stress radiographs with the Telos(R) device and the maximal manual test with the KT-1000 arthrometer. The clinical results were assessed by the IKDC (International Knee Documentation Committee) and OAK (Orthopadische Arbeitsgruppe Knie) scores. The posterior drawer test, varus stress test, posterolateral drawer test and dial test in 30 degrees and 90 degrees flexion were performed for a physical examination. RESULTS: The average side to side difference of the posterior tibial translation in stress radiographs with the Telos(R) device decreased from 10.4+/-2.0 mm to 2.2+/-1.0 mm. The average side to side difference in the maximal manual test with the KT-1000 arthrometer also decreased from 8.2+/-1.5 mm to 1.9+/-1.0 mm. The final IKDC score was A in eleven (21.2%), B in thirty-five (67.3%) and C in six (11.5%) patients. The average OAK score improved from 64.3+/-8.9 to 90.8+/-7.2. CONCLUSION: Tensioning of the laxed remnant PCL with a reconstruction of the AL bundle in chronic PCL injuries showed good clinical results and excellent posterior stability.
Allografts ; Autografts ; Exercise Test ; Humans ; Inlays ; Knee ; Physical Examination ; Posterior Cruciate Ligament* ; Supine Position ; Tendons

While endovascular aneurysm repair (EVAR) is prevailing for the treatment of abdominal aortic aneurysm (AAA) in modern vascular practice, PURPOSE: we conducted nationwide questionnaire survey to investigate the current status of AAA treatment and their results in Korea. METHOD: We reviewed the replies from 28 hospitals (33 departments) to the questionnaire inquiring annual number, clinical features, mode of treatment and results of AAA patients during the period from Jan. 2000 to Jul. 2004. Results: 980 AAA patients were reported including 292 ruptured AAA (29.8%) and 688 non-ruptured AAA (70.2%). For treatment of AAA, 834 (85.1%) surgical repairs (SRs) and 111 (11.3%) endovascualr aneurysm repairs (EVARs) were performed while 35 patients (3.6%) died of AAA rupture before operation. The locations of AAA were infrarenal in 889 (90.7%), juxtarenal in 62 (6.3%), and suprarenal in 29 patients (3.0%). Among 834 patients undergoing SR, 577 patients (69.2%) had non-ruptured AAAs and 257 patients (30.8%) had ruptured AAAs. Mean operative mortality rate was 4.1% after elective SRs, 30.7% after SR for ruptured AAAs, and 2.3% after EVARs. The reported brand name of stent graft devices were various including domestic custom-made in 56 (50.5%), imported brand in 18 (16.2%) while 37 (33.3%) stent grafts were not reported their brand name. The frequencies of type I and III endoleaks after EVAR were reported 5.8% and 5.8% respectively in 86 patients with an available data. CONCLUSION: SR has been used as a major treatment option in Korea for the treatment of AAA patients while EVAR is increasing. The mortality rate of SR of AAA was comparable to western multi-center trial reports but mortality or morbidity rates of EVAR were unable to know in this questionnaire survey.
Aneurysm ; Aortic Aneurysm ; Aortic Aneurysm, Abdominal* ; Blood Vessel Prosthesis ; Endoleak ; Humans ; Korea* ; Mortality ; Questionnaires* ; Rupture ; Treatment Outcome

The hypermethylation of the CpG islands is a common mechanism for the inactivation of tumor-related genes. In the present study, we analyzed the methylation status of genes for cell repair such as hMLH1, MGMT, and GSTP1, and a gastric cancer-specifically methylated DNA fragment, MINT 25 in gastric cancer cases and control groups. The study population consisted of 100 gastric cancer patients (50 distal and 50 proximal carcinomas), and 238 healthy controls. All genes showed more frequent hypermethylation in the cases than in the control group (p<0.0001). We investigated the association between promoter hypermethylation and relevant parameters including age, gender, alcohol consumption, smoking, and family history. There was a common hypermethylation of hMLH1 (p=0.008), MGMT (p= 0.0001), and GSTP1 (p=0.0003) in females. This study also demonstrates that hypermethylation was strongly associated with non-drinkers (MGMT, p=0.046 and MINT 25, p=0.049) and non-smokers (hMLH1, p=0.044; MGMT, p=0.0003; MINT 25, p=0.029). Moreover, the frequency of MINT 25 hypermethylation increased with age (p=0.037), and MGMT methylation was frequently detected in distal gastric cancer than in proximal type (p=0.038). Our study suggested that promoter hypermethylation of the genes involved in cell repair system and MINT 25 is associated strongly with some subgroups of primary gastric carcinoma.
Adult ; Aged ; *DNA Methylation ; Female ; Glutathione Transferase/genetics ; Humans ; Isoenzymes/genetics ; Male ; Middle Aged ; Neoplasm Proteins/genetics ; Nuclear Proteins/genetics ; O(6)-Methylguanine-DNA Methyltransferase/genetics ; Promoter Regions (Genetics) ; Research Support, Non-U.S. Gov't ; Stomach Neoplasms/*genetics

Hypermethylation of CpG island is a common mechanism for the inactivation of tumor-related genes. In the present study, we analyzed 13 genitourinary cancer cell lines for aberrant DNA methylation of 5 tumor-related genes using methylation- specific polymerase chain reaction (MSP). GSTP1 was methylated in 5 (38.5%), E-cadherin in 1 (8%), VHL in 1 (8%), and MGMT and hMLH1 in none (0%). Six out of thirteen genitourinary cancer cell lines had methylation of at least one of five genes; 5 had one gene methylated, and, 1 had two genes methylated. Methylation of these 5 genes was not detected in any of the bladder cancer cell lines. GSTP1 was methylated in all of the 3 prostate cancer cell lines. We conclude that aberrant hypermethylation may be an important mechanism for the inactivation of cancer-related genes in kidney and prostate cancer cell lines.
Bladder Neoplasms/genetics ; Cadherins/genetics ; *DNA Methylation ; DNA Primers ; Genetic Screening/methods ; Glutathione Transferase/genetics ; Human ; Isoenzymes/genetics ; Kidney Neoplasms/genetics ; Ligases/genetics ; Male ; Neoplasm Proteins/genetics ; O(6)-Methylguanine-DNA Methyltransferase/genetics ; Polymerase Chain Reaction ; Prostatic Neoplasms/genetics ; Tumor Cells, Cultured ; Urogenital Neoplasms/*genetics

Germ-line mutations at DNA repair loci confer susceptibility to colon cancer in hereditary non-polypopsis colorectal cancer. Somatic loss of DNA mismatch repair gene has been reported in a large variety of other tumor types. Replication errors(RERs) judged by microsatellite instability(MSI) and its associated mutations have been recognized as an important mechanism in various tumor types. To investigate associations between MSI and oral squamous cell carcinoma, the frequency of MSI using 12 microsatellite markers were analyzed for the series of oral tumors. Of 17 tumors, 8 cases(47%) did not show instability at any of the 12 loci; 5(29%) showed instability at 2~3 loci; and 4(24%) showed instability above 4 loci. The 4 cases showing widespread MSI did not differ from those without evidence of instability in terms of age at diagnosis, degree of differentiation, metastasis to lymph node, tumor location or the presence of mutations in the p53 tumor suppressor gene. DCC and D17S 796 were the most frequently detected in MSI analysis. There were no correlation between smoking and MSI frequency, instead, smoking was suggested to increase the mutation rate of p53 and development of oral carcinomas.
Carcinoma, Squamous Cell* ; Colonic Neoplasms ; Colorectal Neoplasms ; Diagnosis ; DNA Mismatch Repair ; DNA Repair ; Genes, p53* ; Genes, Tumor Suppressor ; Germ-Line Mutation ; Lymph Nodes ; Microsatellite Instability* ; Microsatellite Repeats* ; Mutation Rate ; Neoplasm Metastasis ; Smoke ; Smoking

The objective of this study was to characterise the pattern of p53 mutations in bladder tumor. In this study, 25 bladder transitional cell carcinomas were analyzed by immunohisochemistry (IHC) for p53 nuclear overexpression, and the results were compared with those of polymerase chain reaction-single strand conformational polymorphism (PCR-SSCP) analysis in exon 5-8 of the p53 gene and DNA sequencing analysis. 15 out of 20 cases (75%) showed p53 nuclear immunoreactivities on IHC. On PCR-SSCP analysis, 10 out of 25 cases (40%) had abnormal shifts in mobility. 62% of the mutations were in exon 8. Direct DNA sequencing analysis were performed in these 10 cases to confirm the presence of mutated p53 genes and to determine the type of mutations. Sixteen point mutations were detected in 10 cases. Two specimens had double mutations and another two had triple mutations. G:C-->A:T transitions were the most frequent patterns (62.5%). One mutation was a premature stop codon and two were silent mutations. Three out of 10 had a point mutation at codon 285 (GAG/Glu-->AAG/Lys) and two had at codon 280 (AGA/Arg-->AAA/Lys). One of 16 mutations was transition at hot spot codon 273 with CpG site. These results suggest that altered expressions and point mutations of p53 occured in all grade of bladder cancer, but are more associated with high grade bladder tumors. To elucidate the carcinogenesis of bladder cancer, further studies should be carried out.
Carcinogenesis ; Carcinoma, Transitional Cell* ; Codon ; Codon, Nonsense ; Exons ; Genes, p53 ; Humans* ; Point Mutation* ; Sequence Analysis, DNA ; Urinary Bladder Neoplasms ; Urinary Bladder*

Although bladder transitional cell carcinoma (TCC) is common, the underlying molecular events remain ill-defined. So we attempted to define the role of tumor suppressor genes in the pathogenesis of bladder tumor through a molecular genetic study. For 15 bladder TCC (6 gradeII, 1 gradeIII, and 8 grade IV), we performed the restriction fragment length polymorphism (RFLP) analysis for 6 loci of suspected or established tumor suppressor regions (3p21, 3p24-25, llp15, 13q14, and 17p13). Our data confirm that allelic losses are highly common in bladder tumors. We found that alleles from each of the four chromosomal arms tested were lost in most of the tumors. Reduction of allele occured at 3p21 (13%), 3p24~25 (50%), and 13q14 (38%). However, the greatest frequency of allelic loss was seen for 17p 13 (100% of informative cases) and llp15.5 (87% informative cases). Severe allelic losses of chromosome 17p and pADJ762 on lip were seen only in grade IV, not in grade II. Amplification of 3p21 was seen six out of eight. Amplification of 3p21 has not been previously observed on the other study. Addition to this, we observe the loss of H-ras allele on 11p in one case which was associated with duplication of the retained allele as was demonstrated in Wilms'tumors. The results of out study suggest that deletions of pADJ762 on chromosome 11p and 17p13 occur in high grade bladder tumor and may contribute to the progression of this disease. But, there was no apparent correlation between tumor grade and the loss of 3p or 13q14 alleles although they had some deletions. The role of these genetic alterations in the prognosis of bladder transitional cell carcinoma will require additional follow-up and further studies.
Alleles ; Arm* ; Carcinoma, Transitional Cell ; Chromosomes, Human, Pair 11* ; Genes, Tumor Suppressor ; Lip ; Loss of Heterozygosity ; Molecular Biology* ; Polymorphism, Restriction Fragment Length ; Prognosis ; Urinary Bladder Neoplasms* ; Urinary Bladder*

No abstract available.
Erythrocytes*