Study on the Effect of Porcine Placental Extract Ingestion on Symptoms of Late-onset Hypogonadism Syndrome in Middle-aged and Elderly Men / 日本補完代替医療学会誌

Purpose: An open-label, single-arm study was conducted to evaluate the effectiveness of porcine placental extract (PPE) ingestion to improve late-onset hypogonadism (LOH) syndrome symptoms. In addition, we reported the safety of continuous PPE ingestion in biochemistry tests.Materials and Methods: Test food containing PPE was given daily to healthy male subjects with an Aging Males’ Symptoms (AMS) score of ≥27, who were assessed as having LOH syndrome. AMS score, blood hormone level, and other parameters were measured before the start of PPE ingestion, Week 4, and Week 8. And adverse events that occurred during the ingestion period were accumulated.Results: Excluding one subject who withdrew from the study due to an adverse event unrelated to the test food, 14 subjects were included in the analysis. In terms of AMS score, no improvement was observed at Week 4, whereas a significant improvement was observed at Week 8. AMS scores were further categorized into three subscales (psychological, somatic, and sexual) and analyzed. The results showed a significant improvement from baseline in somatic scores at Week 8. Meanwhile, no significant change in sexual hormone levels was observed. At Week 8, improvements from baseline were observed in renal function and LDL cholesterol level. There were no adverse events related to the test food.Conclusion: The study demonstrated that 8-week PPE ingestion in middle-aged and elderly men can improve LOH syndrome symptoms while having little effect on sexual hormone secretion.

<b>Hochuekkito Efficacy in Late-Onset Hypogonadism (LOH) Patients </b> / 日本東洋医学雑誌

The purpose of this study is to evaluate the efficacy of hochuekkito for late-onset hypogonadism (LOH) patients. We administered hochuekkito 7.5 g/day for 8 weeks to 47 patients with LOH whose AMS scale was more than 27. We assessed the patients' symptom change with the AMS, SHIM, SDS, BDI, and SF-36. We measured their endocrine profiles and levels of their cytokines. At the end of study, 31 of 47 patients were evaluable. No significant difference in subjective symptoms was seen with any questionnaire after 8 weeks hochuekkito administration. However, hochuekkito significantly increased free testosterone and decreased ACTH/cortisol levels. Thus we believe hochuekkito is beneficial for the treatment of LOH.

Genomic instability at the 13q31 locus and somatic mtDNA mutation in the D-loop site correlate with tumor aggressiveness in sporadic Brazilian breast cancer cases

OBJECTIVE: Genomic instability is a hallmark of malignant tissues. In this work, we aimed to characterize nuclear and mitochondrial instabilities by determining short tandem repeats and somatic mitochondrial mutations, respectively, in a cohort of Brazilian sporadic breast cancer cases. Furthermore, we performed an association analysis of the molecular findings and the clinical pathological data. METHODS: We analyzed 64 matched pairs of breast cancer and adjacent non-cancerous breast samples by genotyping 13 nuclear short tandem repeat loci (namely, D2S123, TPOX, D3S1358, D3S1611, FGA, D7S820, TH01, D13S317, D13S790, D16S539, D17S796, intron 12 BRCA1 and intron 1 TP53) that were amplified with the fluorescent AmpFlSTR Identifiler Genotyping system (Applied Biosystems, USA) and by silver nitrate staining following 6% denaturing polyacrylamide gel electrophoresis. Somatic mtDNA mutations in the D-loop site were assessed with direct sequencing of the hypervariable HVI and HVII mitochondrial regions. RESULTS: Half of the cancer tissues presented some nuclear instability. Interestingly, the D13S790 locus was the most frequently affected (36%), while the D2S123 locus presented no alterations. Forty-two percent of the cases showed somatic mitochondrial mutations, the majority at region 303-315 poly-C. We identified associations between Elston grade III, instabilities at 13q31 region (p = 0.0264) and mtDNA mutations (p = 0.0041). Furthermore, instabilities at 13q31 region were also associated with TP53 mutations in the invasive ductal carcinoma cases (p= 0.0207). CONCLUSION: Instabilities at 13q31 region and the presence of somatic mtDNA mutations in a D-loop site correlated with tumor aggressiveness.

Effects of mutation in Cosmc non-coding region on the transcription level of Cosmc mRNA in Tn antigen positive tumor cells / 中华微生物学和免疫学杂志

Objective To explore the mutation in corel β3-galactosyl-transferase specific molecular chaperone(Cosmc、no-coding region and it's effects on the transcription level of Cosmc in Tn antigen positive tumor cells.Methods The Tn antigen positive(Tn+)and negative(Tn-)cells were separated from tumor tissues by immune magnetic bead,then the genomic DNA(gDNA),total RNA were prepared by Qiagen AllPrep DNA/RNA mini kit. In these cells.the transcription levels of T-synthase and Cosmc mRNA were tested by RT-PCR.the DNA of Cosmc non-coding region was amplified by PCR,the mutation in Cosmc non-coding region were further detected by sequencing.Results There are no mutation appearing in Tn-cells,one or more mosaic sequence allele appearing in portion of patient's Tn-cells.Almost of the Tn+cells which separated from tumor tissues and Jurkat T cell exists mutation.but the mutation style and mutation point were not saine in different tumor.Thtee patient's Tn+cells have loss of hetemzygosity(LOH),four patient's Tn+cells and Jurkat T cell have point mutation.Although no difference of transcription level of T-synthase mRNA in Tn+ and Tn-cells.but the transcription level of Cosmc mRNA in Tn+ cell was much lower than that in Tn-cell.The ratio of T-synthase/Cosmc mRNA in Tn+ tumor cells was hiigher than that in Tn-cell.Conclusion The tumor Tn antigen arise from mutation in Cosmc non-coding region maybe result from transcription level decreased of Cosmc mRNA.

Genetic instability of P57~(kip2) misexpression in human hepatocellular carcinoma / 基础医学与临床

Objective To study the expressions of P57~(kip2) mRNA and genetic instability of P57~(kip2) in human hepatocellular carcinoma (HCC).Methods In situ hybridization(ISH) was used to detect the expression of P57~(kip2) mRNA in HCC.MSI and LOH were detected by PCR-polyacrylamide gel electmphoresis-silver staining method.Results There was no expression of P57~(kip2) mRNA found in normal liver tissue.The expression rate of P57~(kip2) mRNA in both pericancerous cirrhosis and hepatocellular carcinoma was 26.7%(8/30).LOH was not identified in 30 cases on three microsatellite loeies;there were 2 microsatellite loeies showing MSI,the total rate of MSI was 16.7%.There was correlation found between the MSI of D11S1760 locies and the expression of P57~(kip2) mRNA(P＜0.05).Conclusion The disorder expression of P57~(kip2) mRNA indicated that P57~(kip2) might be involved in hepatocarcinogenesis and prognosis.MSI may be one of the reasons that explains the disorder expression of P57~(kip2) mRNA in hepatocarcinogenesis and prognosis.

Absence of PRKAR1A loss of heterozygosity in laser-captured microdissected pigmented nodular adrenocortical tissue from a patient with Carney complex caused by the novel nonsense mutation p. Y21X / Ausência da perda de heterozigose do PRKAR1A em células capturadas por microdissecção a laser de tecido de nódulo pigmentoso adrenocortical de um paciente com complexo de Carney causado por uma nova mutação nonsense

OBJECTIVE: Primary pigmented nodular adrenocortical disease (PPNAD) is the main endocrine manifestation of Carney complex, a multiple neoplasia syndrome caused by PRKAR1A gene mutations. The presence of PRKAR1A loss of heterozygosity (LOH) in adrenocortical tumorigenesis remains controversial. The aim of the present study is to investigate the presence of PRKAR1A LOH in adrenocortical cells in a patient with Carney complex. METHODS: The LOH was investigated using a PRKAR1A informative intragenic marker by GeneScan software analysis in DNA obtained from laser-captured microdissected cells of several adrenal nodules. Patients: A young adult male patient with Carney complex and his family were studied. RESULTS: A novel heterozygous mutation (p. Y21X) was identified at PRKAR1A in blood DNA of the male proband and his relatives. No PRKAR1A LOH was evidenced in the laser-captured microdissected cells from PPNAD tissue by different methodologies. CONCLUSION: We identified a new PRKAR1A nonsense mutation and in addition we did not evidence PRKAR1A LOH in laser-captured nodules cells, suggesting that adrenocortical tumorigenesis in PPNAD may occurs apart from the second hit.

OBJETIVO: A doença adrenocortical nodular pigmentosa primária (PPNAD) é uma das manifestações do complexo de Carney, uma neoplasia endócrina múltipla causada por mutações no PRKAR1A. A perda de heterozigose (LOH) do PRKAR1A na tumorigenese adrenal permanece controversa dada à possibilidade de contaminação com o tecido normal. Nosso objetivo foi investigar a presença de LOH no PRKAR1A a partir de células do nódulo adrenal de um paciente com complexo de Carney. MÉTODOS: A pesquisa da LOH do PRKAR1A foi realizada através do estudo de um marcador intragênico em DNA de células do nódulo adrenal microdissecadas a laser, evitando contaminação com o tecido normal. Pacientes: Um paciente com PPNAD e cinco familiares foram estudados. RESULTADOS: A nova mutação (p. Y21X) foi identificada no PRKAR1A sem evidência de LOH no tecido adrenal. CONCLUSÃO: Identificamos uma nova mutação no PRKAR1A e não evidenciamos LOH nas células dos nódulos adrenocorticais, sugerindo que a PPNAD possa ocorrer na ausência de um segundo evento molecular.

The Expression of Human Tumor Suppressor Gene beclin 1 is Down-regulated in Gastric and Colorectal Cancer / 生物化学与生物物理进展

Human tumor suppressor gene beclin 1 regulates cell growth through autophagy. The mRNA expression of beclin 1 was reported to be down-regulated in breast cancer with high frequency of loss of heterozygosity (LOH). However, there was no report about the expression levels or the regulatory mechanisms of beclin 1 in gastric and colorectal cancer. Both the mRNA and protein expression levels of beclin 1 was detected in the tissues of gastric and coiorectai cancers, as well as the aberrant DNA methylation and LOH related to the expression of beclin 1. By comparing with normal tissues adjacent to the tissue of these tumors, it was found that beclin 1 mRNA expression levels were significantly decreased in gastric tumor tissue. Furtherly by explorating the 5' region of beclin 1 gene sequence, a large and dense CpG island was discovered and meanwhile methylations in the promoter and the intron 2 regions of beclin 1 were found in both gastric and colorectal tumors. And LOH was found in gastric tumors. These findings suggested that aberrant DNA methylation, as well as LOH, were involved in the regulation of beclin 1 expression in gastric and colorectal cancer.

Loss of heterozygosity of E-cadherin in endometrial carcinomas and endometrial hyperplasias / 대한산부인과학회지

OBJECTIVE: Microsatellite alteration such as loss of heterozygosity (LOH) has been reported to be a novel mechanism for the inactivation of tumor suppressor gene and related to carcinogenesis in many malignant tumors. E-cadherin protein coded by gene on chromosome 16q22.1 may play a principal role for tumor suppression. However LOH of E-cadherin has been rarely studied in endometrial carcinoma. The purpose of this study was to investigate the loss of heterozygosity (LOH) of E-cadherin in endometrial carcinoma and endometrial hyperplasia and to correlate their results with various clinicopathological factors. METHODS: LOH of E-cadherin on the chromosome 16q22.1 analysis was performed by using polymerase chain reaction (PCR) for three polymorphic microsatellite markers (D16S419, D16S3106, D16S498) and automatic laser fluorescent DNA sequencer in 30 cases of endometrioid endometrial adenocarcinomas and in 20 cases of endometrial hyperplasias. The relationship between LOH of E-cadherin and clinical profile was analyzed. RESULTS: Increased LOH of E-cadherin was found in endometrial carcinomas (50%) compared to endometrial hyperplasias (5%) (P=0.01, Fisher's exact test). The incidence of LOH of E-cadherin in endometrial carcinomas also showed significantly lower in stage below Ia. (P=0.034, Fisher's exact test) LOH of E-cadherin was not associated with histologic grade and lymph node metastasis. (P=0.42, P=0.5, Fisher's exact test) CONCLUSIONS: These results suggests that LOH of E-cadherin may contribute to the development of endometrial carcinoma, especially in above stage Ib.

Loss of Heterozygosity of Major Tumor Suppressor Genes in Invasive Ductal Carcinomas / 한국유방암학회지

PURPOSE: Breast cancer is one of the most frequent malignant tumors in Korea. The major tumor suppressor genes (TSGs) such as p16, Rb, E-cadherin and p53 may play important roles in cell cycle regulation, apoptosis and the regulation of the expression of other genes as well as tumor suppression. Microsatellite alteration such as loss of heterozygosity (LOH) have been reported to be a novel mechanism of carcinogenesis and a useful prognostic factor for many malignant tumors. Also, LOH is also known to be related with allelic loss of various TSGs. This study evaluated LOH of 4 TSGs in invasive ductal carcinomas (IDCs) and we correlated these results with the clinicopathological factors. METHODS: LOH analysis was carried out using a polymerase chain reaction with 12 polymorphic microsatellite markers of 4 TSGs in 50 surgically resected tumors and their non-tumorous counterparts. RESULTS: There was no detectable LOH in the normal tissue. LOH was detected in 86% of the 50 cases of IDCs. LOH was detected on all chromosomes and this showed a statistical difference between benign tumor and malignant tumor. LOH of p16, Rb, E-cadherin and p53 TSGs was detected in 36%, 26%, 54% and 60% of the tumors, respectively. LOH of the p16 and Rb genes was inversely correlated with tumor grade 1. The low rate of detecting LOH on the E-cadherin gene was noted in T1 tumor and stage I disease. LOH of the p53 gene correlated well with the tumor size and stage. The LOH-High results correlate well with the tumor size and stage and the LOH-High results are similar to those of the p53 gene LOH. CONCLUSION: These results suggest that LOH of the 4 major TSGs may contribute to the development and invasion of IDCs. Also, the combined use of various LOH markers may help in deciding the prognosis of IDCs.

Genetic alteration and expression of ING1 in human lung carcinoma / 肿瘤

Objective: To detect the loss of heterozygosity (LOH) of ING1 gene microsatellite and the expression of p33ING1b protein in lung carcinoma and to investigate their association with the carcinogenesis of lung cancer. Methods: Seventy cases of fresh-frozen lung cancers and 217 cases formalin-fixed, paraffin-embedded specimens were examined. The LOH frequency of ING1 gene microsatellite was detected by polymerase chain reaction-single strand conformation polymorphisms (PCR-SSCP) method. The expression of p33ING1b protein was determined by immunohistochemical method and tissue microarray technology. Results: The total LOH frequency of four microsatellites of ING1 gene was 55.7% (39/70) in lung carcinomas. The LOH had higher frequency if it was closer to the ING1 locus. But the LOH frequency had no correlation with clinicopathological parameters. The total LOH frequency of p33ING1b protein was 47.0% (102/217) in lung carcinomas. The positivity of p33ING1b in squamous carcinoma was significantly higher than that in adenocarcinoma (AC), adenosquamous carcinoma (AdCa), and bronchioloalveolar carcinoma (BAC) (P <0.05), while there was no significant difference between AC, AdCa, and BAC. The expression of p33ING1b protein was not associated with the clinicopathological characteristics but a negative correlation with LOH frequency was observed (P <0.05). Conclusion: There existes higher frequency of low expression or deletion of ING1 protein in lung carcinoma tissues. The LOH of ING1 gene microsatellites also have a higher frequency. LOH is one of the important reasons for the abnormal expression of ING1 gene, which causes the down-regulation of ING1 gene and inactivation of ING1 protein. Finally ING looses its inhibitory effect on the growth of tumor cells and results in carcinogenesis of lung cancer.

Estudos sobre a aplicabilidade da citopatologia no diagnóstico precoce do câncer bucal / Studies on the applicability of cytopathology in the early diagnosis of oral cancer

O câncer bucal é uma doença que afeta principalmente homens acima de 40 anos com hábito de fumar e beber. Em geral, esses pacientes têm o diagnóstico da enfermidade realizado tardiamente, acarretando tratamentos complexos, mutilantes e de alto custo tanto financeiro como social e de prognóstico sombrio a ponto do câncer bucal apresentar uma taxa de sobrevida em 5 anos inferior a 50%. O diagnóstico precoce representa uma alternativa importante para alterar esse panorama. Os recursos de que os profissionais da área da saúde, principalmente os cirurgiões-dentistas, dispõem são o diagnóstico clínico de lesões estabelecidas acompanhado da biópsia com exame histopatológico.Outra alternativa para o diagnóstico precoce dos carcinomas espinocelulares ocorre quando estes são precedidos de lesões cancerizáveis. Nas duas últimas décadas, a utilização da citopatologia como método de diagnóstico de danos celulares prévios ao aparecimento de lesões clínicas possibilitou sua utilização em mucosa bucal. Os estudos apresentados tratam especificamente desta aplicação clínica, abordando: a) os aspectos genéticos do câncer bucal; b) a correlação histocitopatológica de lesões cancerizáveis em câncer bucal e na mucosa normal exposta aos carcinógenos; c) a validação das amostras citopatológicas; d) a padronização da técnica de extração de DNA em células esfoliadas da mucosa bucal...

Oral cancer affects mainly men over 40 years who are exposed to tobacco and alcohol. These patients usually receive a late diagnosis, which results in complex, mutilating treatments with high financial and social costs, poor prognosis, and, therefore, a 5-year survival rate lower than 50%. Early diagnosis is critical to change this situation. The resources currently available to healthcare professionals, particularly dentists, are the clinical diagnosis of lesions and biopsies for histopathologic examination. Another alternative for the early diagnosis of squamous cell carcinomas is the identification of precursor lesions. In the last two decades, cytopathology has been used as a method to diagnose cell damage that precedes the appearance of clinical lesions, and this use can be extended to the oral mucosa. The studies reported here deal specifically with this clinical application, and discuss: A) the genetic factors in oral cancer; B) the cytohistologic correlation of cancer precursor lesions and the normal mucosa exposed to carcinogens; C) the validation of cytopathologic samples; D) the standardization of the technique to extract DNA of cells exfoliated from the oral mucosa...

Expression and Mutation Analysis of RKIP (Raf-1 Kinase Inhibitor Protein) in Human Gastric Cancer

PURPOSE: RKIP (Raf kinase inhibitor protein) is a novel candidate tumor suppressor, known to inhibit the MAPK signaling by interfering with the MEK phosphorylation by Raf-1. The aim of this study was to investigate the expression of RKIP and analyze the pattern of inactivation and mutation of the RKIP gene in human gastric cancer. METHODS: To explore if RKIP inactivation is implicated in gastric tumorigenesis, an expression analysis on the transcription and protein expression levels and a mutational analysis of RKIP were performed in 15 human gastric cancer cell lines and 92 primary carcinoma tissues. RESULTS: Abnormal reduction of the level of RKIP expression was frequently detected in the cancer cell lines and primary tumor tissues, at both the transcript and protein levels. Moreover, the expression level of RKIP in the tumor cells was inversely correlated with the level of Erk phosphorylation, indicating that RKIP plays a key role in the regulation of the Raf-MEK-Erk signaling pathway in human gastric cells. While the expression of the RKIP transcript was not re-activated in low expressor cells by treatment with the demethylating agent 5'Aza-dC, the genomic RKIP was detected at low levels in many cancer cell lines, suggesting that an abnormal reduction of level of RKIP expression in tumors might be caused by allelic deletion of the gene rather than transcriptional silencing due to aberrant DNA hypermethylation. A loss of heterozygosity study, using an intragenic polymorphic marker, revealed that approximately 21% of the gastric cancers harbored allelic loss of the RKIP gene. CONCLUSION: Collectively, this study has demonstrated that RKIP is a tumor suppressor, whose expression is frequently downregulated by allelic deletion in human gastric cancers. This study also suggests that an altered expression of RKIP might contribute to the development of gastric cancer via abnormal elevation of the Raf-Erk signaling pathway.

EBP50:a Novel Cancer Suppressor Protein / 生物化学与生物物理进展

EBP50(ERM-binding phosphoprotein-50),a multifunctional adapter protein with 358 amino acids and two PDZ domains, regulates cell growth and migration. Lines of evidences indicate that it is a potential cancer suppressor protein. Loss of heterozygosity (LOH) and intragenic mutation of the ebp50 gene have been found in both primary breast tumors and breast cancer cell lines. EBP50 suppresses the breast cancer cell proliferation via its interaction with many tumor suppressor protein including PTEN, SYK, MERLIN, etc. Here the molecular structure of EBP50, signal pathway regulated by EBP50, and the relationship between breast cancer development and EBP50 are discussed.

Loss of Heterozygosity of E-Cadherin Gene and Protein Expression in Invasive Ductal Carcinomas / 한국유방암학회지

PURPOSE: The E-cadherin gene, located on chromosome 16q22, may play principal roles in cell adhesion with the loss of E-cadherin expression leading to a propensity for a great number of malignant properties. The loss of heterozygosity (LOH) on 16q22 has rarely been studied in invasive ductal carcinomas. Our objectives were to evaluate the LOH of E-cadherin and the protein expression in invasive ductal carcinomas and their correlation with various clinicopathological factors. METHODS: The LOH analysis was performed using polymerase chain reactions with three polymorphic microsatellite markers (D16S419, D16S3106 and D16S498) in 50 surgically resected tumors and their non-tumorous counterparts. The E-cadherin protein expression was studied using immunohistochemistry. RESULTS: The LOH and loss of protein expression were detected in 54% and 46% of the tumors, respectively. There was no LOH or protein loss detected in the non-tumor lesions. The LOH results were well correlated with the tumor size and lymph node metastasis. The protein loss results were well correlated with tumor histological grade. No correlation was found between LOH and protein loss. CONCLUSION: These results suggest that the LOH of E-cadherin may be associated with tumor metastasis and tumor progression and E-cadherin protein loss may be related with the dedifferentiation in some portions of invasive ductal carcinomas. We propose the LOH of E-cadherin and protein loss may contribute to tumor progression by independent mechanism.

A Study of the Oncological Characteristics Affecting the Growth Patterns of the Gastric Cancer

PURPOSE: In general, the depth of tumor invasion correlates with lymph node metastasis in gastric cancer, but some cases show a discrepancy between the tumor depth and nodal status. Therefore, this study was designed to investigate the factors affecting tumor growth patterns including the depth of invasion and lymph node metastasis. METHODS: The clinico-pathological characteristics of 1048 gastric cancer, from 1993 to 1999, were investigated. These cancer cases were divided into three groups: the early gastric cancer with lymph node metastasis (EP), the serosa infiltrated advanced cancer with no lymph node metastasis (SN), and the serosa infiltrated advanced gastric cancer with lymph node metastasis (SP) groups. The expression status of the proteins related with tumor growth, including matrix metalloproteinase-2 and 9 (MMP-2 and MMP-9), VEGF, nm23 and p53, were compared using immunohistochemical stain, p53 gene mutation, Microsatellite instability (MSI) and Loss of heterozygosity (LOH). RESULTS: There were 35 (3.4%) of the total cases in the EP group, and 10.1% in Early gatric cancer cases. The female ratio in the EP group was higher than in the SN group but this was not statistically significant. The most common gross types in the EP and SN groups were IIc and Borrmann type III, respectively, and the mean tumor size was largest in the SN group. The cell differentiation of the SN group was better than that in the EP group (68.6 vs. 41.2%). The levels of MMP-2 and -9 were higher in the SN and SP groups than those in the EP group. However, the levels of VEGF and nm23 between the groups were no different. The EP group had the highest degree of p53 mutation. There was no difference in the MSI and LOH expression status between the groups. CONCLUSION: There were some different growth patterns in the gastric cancers between the lymph node metastasis dominant (EP) and depth of infiltration dominant (SN) groups. MMP-2 and -9, tumor differentiation and mutated p53 gene exon may correlated with the tumor growth pattern. Further study is suggested to find the difference between the EP and SN groups, which could provide information on which factors determine nodal metastasis or the depth of a infiltration dominant growth pattern.

Loss of heterozygosity of microsatellite DNA on 6q in bladder tumor / 中国康复理论与实践

@#ObjectiveTo investigate the loss of heterozygosity (LOH) on 6q in bladder tumor.MethodsD6S404 and D6434 microsatellite markers near 6q21 were tested by PCR-SSLP-stain method on tumor DNA from 31 cases of bladder tumor.ResultsAmong these 31 cases of bladder tumor,LOH was detected in tumor tissues on site for D6S404 (35.5%) and D6S434(22.6%).ConclusionOne or more tumor suppressor gene near 6q21 maybe relevant for the development of bladder tumor.

Identification of Tumor Suppressor Loci on the Long Arm of Chromosome 15 in Primary Small Cell Lung Cancer

Small cell lung cancer (SCLC) frequently shows a loss of heterozygosity (LOH) on chromosome 15q. In order to define the commonly affected region on chromosome 15q, we tested 23 primary SCLCs by microsatellite analysis. By analyzing 43 polymorphic microsatellite markers located on chromosome 15q, we found that 14 (60.8%) of 23 tumors exhibited a LOH in at least one of the tested microsatellite markers. Two (14.3%) of the 14 tumors were found to have more than a 50% LOH on chromosome 15q. LOH was observed in five commonly deleted regions on 15q. Of those regions, LOH from D15S1012 to D15S1016 was the most frequent (47.8%). LOH was also observed in more than 20-30% of tumors at four other regions, from D15S1031 to D15S1007, from D15S643 to D15S980, from D15S979 to D15S202, and from D15S652 to D15S642. Four of the 23 tumors exhibited shifted bands in at least one of the tested microsatellite markers. Shifted bands occurred in 3.2% (29 of 914) of the loci tested. Our data suggests the presence of at least five tumor suppressor loci on chromosome 15q in SCLC, and further that these may play an important role in SCLC tumorigenesis.

Identification of Tumor Suppressor Loci on the Short Arm of Chromosome 16 in Primary Small Cell Lung Cancers / 결핵

BACKGROUND: Loss of the short arm of chromosome 16 is a frequent event in various cancers, which suggests the presence of tumor suppressor gene(s) there. To map precise tumor suppressor loci on the chromosome arm for further positional cloning efforts, we tested 23 primary small cell lung cancers. METHODS: The DNAs extracted from paraffin embedded tissue blocks with primary tumor and corresponding control tissue were investigated. Twenty polymorphic microsatellite markers located in the short arm of chromosome 16 were used in the microsatellite analysis. RESULTS: We found that six (26.1%) of 23 tumors exhibited LOH in at least one of tested microsatellite markers. Two (8.7%) of 6 tumors exhibiting LOH lost a larger area in chromosome 16p. LOH was observed in five common deleted regions at 16p. Among those areas, LOH between D16S668 and D16S749 was most frequent (21.1%). LOH was also observed at four other regions, between D16S3024 and D16S748, D16S405, D16S420, and D16S753. Six of 23 tumors exhibited shifted bands in at least one of the tested microsatellite markers. Shifted bands occurred in 3.3% (15 of 460) of the loci tested. CONCLUSION: Our data demonstrated that at least five tumor suppressor loci might exist in the short arm of chromosome 16 and that they may play an important role in small cell lung cancer tumorigenesis.

DCC Gene and Protein Expression in Colorectal Cancer

PURPOSE: The germline, or somatic, inactivation of tumor suppressor genes, through point mutation, or deletion, plays an important role in carcinogenesis. Several gene alterations, such as adenomatous polyposis coli (APC), deleted in colorectal cancer (DCC) and p53, have been detected in the development of colorectal cancer. Within these genes, a loss of heterozygosity (LOH) at the DCC gene locus was frequently associated with colorectal tumors, and the LOH of the DCC gene, and the expression of the DCC protein, might be related to malignant formation and metastasis. The aim of this study was to determine the DCC LOH and the expression of DCC protein in colorectal cancers, and evaluate their prognostic value and relationship with the clinicopathological data. MTHODE: Fifty colorectal cancer tissues were obtained from resected specimens. Using formalin-fixed paraffin- embedded sections as a source of DNA, we examined the DCC protein in the tissue through immunohistochemical stainings and immunoblotting analysis, the DCC LOH through a polymerase chain reaction (PCR) and single strand conformation polymorphism (SSCP). RESULTS: DCC LOH was observed in 24 of the 50 patients (48.0%). The expression of the DCC protein was decreased in the cancer tissue (62.3 23.6%) compared with the adjacent normal mucosa inform the immunoblotting analysis. A decreased DCC protein expression was also observed from the immunohistochemistry, which coincided with the immunoblotting analysis. However, both the DCC LOH and the decreased DCC protein were not related to the clinical and pathological parameters, such as location of tumor, tumor size, histological type and the venous, and lymphatic invasions. There were significant correlations between the DCC protein expression and tumor progression, and hematogenous metastasis (P<.05). CONCLUSIONS: A decreased expression of the DCC protein was noted in human colorectal cancers, and there was a significant relationship between the expression of the DCC protein and distant metastasis, but there was no correlation between the DCC LOH and distant metastasis. These results suggest that the expression of the DCC protein might be related to tumor progression and metastatic potential, and the DCC protein immunoreactivity may be a useful prognostic factor in patients with colorectal cancers.

Loss of Heterozygosity in Endometriosis / 대한산부인과학회잡지

OBJECTIVE: Endometriosis is a very common gynecological condition in which tissue similar to endometrium proliferates at sites outside the uterine cavity. Although it generally remain a benign condition, malignant transformation has been documented, and it is commonly found in association with endometrioid subtype ovarian carcinoma. In order to identify the genomic change in those areas possibly involved in the pathogenesis of endometriosis, we performed LOH analysis. METHODS: Twenty seven cases of endometriosis were analyzed for the detection of LOH using 5 microsatellite markers. LOH analysis was performed by PCR, capillary electrophoresis and gene scan analysis using DNA from sections of tumor and normal tissue pairs. RESULTS: Twenty two of 27 (81.5%) cases demonstrated LOH at one or more loci. The frequency of LOH was 37.0% (D18S69), 25.9% (D22S274), 14.8% (D22S283), 7.4% (D6S286), 7.4% (D13S160). CONCLUSION: The frequencies of LOH was increased in higher stage of endometriosis. Most notable findings were found at chromosome 18 and 22 loci (D18S69, D22S274). These region might involve the some candidate genes closely related with the pathogenesis of endometriosis.