Clinical significance of ERG rearrangement subtype and its association with increased p53 expression in Japanese and German prostate cancer.

UNLABELLED: This study investigated differences in prevalence of the androgen-regulated transmembrane protease serine 2 (TMPRSS2) and ETS transcription factor family member, v-ets erythroblastosis virus E26 oncogene homolog (ERG) fusion gene (TMPRSS2-ERG fusions) in clinically localized prostate cancer Japanese and German patients. A total of 105 specimens, including 69 Japanese and 36 German patients, were collected. The status of TMPRSS2-ERG fusion was determined by fluorescence in situ hybridization, and correlations of the TMPRSS2-ERG fusion with clinicopathological characteristics and immunohistochemistry were studied. Gene fusions were identified in 20% (14/69) of Japanese and 53% (19/36) of German patients (P < 0.001). The difference in the type of gene fusion between the two ethnic groups was statistically significant (P=0.024). Overexpression of ERG protein was significantly associated with gene fusion. Biochemical recurrence was significantly higher in patients with ERG overexpression than in those without, and not related to TMPRSS2-ERG fusion status. Interestingly, two types of gene fusions (deletion and increase of copy number) were significantly associated with increased p53 expression (P = 0.005). Association of specific gene fusions harboring higher genomic alterations with p53 expression levels suggests that p53 mutation might drive more aggressive arrangements of TMPRSS2-ERG fusion in prostate cancer. KEYWORDS: ERG, p53, prostate cancer, TMPRSS2-ERG fusion.

[Complete immunization coverage and reasons for non-vaccination in a periurban area of Abidjan]. / Couverture vaccinale complete des enfants de 12 a 59 mois et raisons de non-vaccination en milieu periurbain abidjanais en 2010.

An immunization coverage survey was conducted among children aged 12-59 months in a suburban neighbourhood in Abidjan. The objective was to determine the complete immunization coverage, the reasons for non-vaccination and factors influencing the immunization status of children. The method of exhaustive sampling enabled us to interview the mothers of 669 children using a questionnaire. Overall vaccination coverage was 68.6% with 1.2%, with 1.2% of children never having received vaccine. The logistic regression analysis showed that the level of education, knowledge of the immunization schedule and the marital status of mothers, as well as the type of habitat, were associated with full immunization of children. These determinants must be taken into account to improve vaccination coverage.

[Anaemia, iron index status and acute phase proteins in malaria (Abidjan, Côte d'Ivoire)]. / Anémie, métabolisme du fer et protéines de la réaction inflammatoire au cours du paludisme (Abidjan, Côte d'Ivoire).

Clinical signs of malaria are the combined expression of several biological mechanisms. During this parasite infection, anaemia can be the consequence of several different pathogenic mechanisms. It can be an acute haemolytic anaemia due to a mechanical and immune action of the parasite or an inflammation. Besides, in Africa malaria matches with iron deficiency area. So, malarial anaemia in tropical area can be a characteristic of iron deficiency The purpose of this survey was to define the features of malarial anaemia and elucidate the link of all biological processes involved. A black population living in tropical urban areas, with fever and diagnosed Plasmodium-infection was assessed. Parasitaemia, haemoglobin, hematocrit, average corpuscular volume and average corpuscular haemoglobin were determined. For each patient, iron index status and acute phase protein were assessed with the plasmatic iron, ferritin, haptoglobin, transferrin and C-reactive protein. Regardless of gender and age, the characteristics of malarial anaemia are microcythaemia and hypochromia. Anaemia occurs as frequently as parasitaemia is high. When parasitaemia is low anaemia gets a haemolytic feature. When parasitaemia is high, anaemia gets haemolytic and inflammatory features. Anaemia occurs more often with a good iron index status.

[Diabetes in Ivory Coast: special epidemiological features]. / Le diabète sucré diagnostiqué en Côte d'Ivoire: des particularités epidémiologiques.

Within less than a quarter century diabetes has become a health problem in developing countries. In Africa this metabolic disorder is found in a wide variety of sometimes atypical forms. The purpose of this study was to highlight the special epidemiological features of medically diagnosed diabetes in Ivory Coast. Data from the files of 10320 African patients who presented at a major national outpatient care centre between January 1, 1991 and December 31, 2000 were compiled and analyzed. Findings showed that morbidity gradually increased from 30 to 49 years then stabilized from 50 to 69 years with a higher rate in males between 30 and 49 years. One of the five national ethnic groups appeared to be most affected and two appeared to be relatively unaffected. On the basis of several criteria, 5968 patients were classified as type 1 in 11.8% of cases, type 2 without excess body weight in 48.7% and type 2 with excess body weight in 39.5%. The second of these identified groups was characterized by intermediate-discovered glycaemia and older age at diagnosis. Epidemiological features included age of occurrence and higher morbidity in young male patients, probable higher premature mortality, likely links with socio-cultural environmental factors and existence of two type 2 subgroups. This profile underlines the challenges of screening, management and prevention of diabetes in Ivory Coast.

The relationship of DNA ploidy to chromosomal instability in primary human colorectal cancers.

The aim of this investigation was to corroborate the relationship between DNA ploidy and chromosomal variation in surgically removed colorectal cancers. For 101 specimens from 21 advanced colorectal cancers, the numerical variations in chromosomes 7, 17, and 18 among cells were measured by fluorescence in situ hybridization using DNA probes specific for centromere of each chromosome, and DNA ploidy was determined by laser scanning cytometry or flow cytometry. DNA diploidy (DNA index = 1.0) was linked with minor variation in copy number of chromosomes 7, 17 and 18, whereas DNA aneuploidy (DNA index > or = 1.2) was found exclusively in tumors with large variations in centromere copy number for all chromosomes. There was a significant difference in the degree of intercellular variations in chromosome copy number between diploid and aneuploid clones for all chromosomes examined (P < 0.001). In near-diploid clones (1.0 < DNA index < 1.2), the numerical variation of chromosome 18 was significantly different from that in diploid clones (P < 0.002), but it was not different from that in aneuploid clones. These observations support the hypothesis that chromosomal instability is associated with DNA aneuploidy in colorectal cancers. Additionally, they suggest that near-diploid tumors are also unstable at a lower level than classic aneuploid tumors and that all chromosomes are not affected equally in near-diploid cases.

Global genomic changes induced by two-stage carcinogen exposure are precancerous alterations in non-transformed human liver epithelial THLE-3 cells.

Global genomic changes, including DNA aneuploidy, may be necessary for carcinogenesis; however, such genomic changes in precancerous cells have not been studied extensively. To identify early global genotypic changes associated with precancerous lesions, a non-transformed human liver epithelial cell line, THLE-3, was treated with benzo[a]pyrene or N-methyl-N-nitro-N-nitrosoguanidine, then by 12-O-tetradecanoyl-phorbol-13-acetate, resulting in morphological transformation of cells. We examined genotypic changes of the transformed cells by laser scanning cytometry, fluorescence in situ hybridization, and comparative genomic hybridization. Transformed fusiform cells displayed tetraploidy, chromosomal instability, DNA copy number aberrations. Cells with these changes were still in the precancerous stage. However, it is suggested that these global genomic changes including tetraploidization provide cells with genetic alterations leading to cancer.

Relationship between chromosomal instability and intratumoral regional DNA ploidy heterogeneity in primary gastric cancers.

The purpose of this study was to elucidate the relationship between intratumoral regional heterogeneity in DNA ploidy and chromosomal instability (CIN) in primary gastric adenocarcinomas. In 45 sporadic gastric adenocarcinomas, we measured DNA ploidy and numerical aberrations for chromosomes 7, 11, 17, and 18 by laser scanning cytometry and fluorescence in situ hybridization, respectively, in small tissue specimens taken from 2 to 6 (on the average 4) different portions of the same tumor. A total of 231 specimens including 45 normal control specimens were examined. All 98 tumor specimens with DNA aneuploidy (DNA index > or = 1.2) showed large intercellular variations in chromosome copy number, indicating CIN. In contrast, 85 tumor specimens with (near) diploidy (1.0 < or = DNA index < 1.2) exhibited much small intercellular variations in chromosome copy number as compared with aneuploid specimens (P < 0.0001). The relationship between DNA ploidy and intercellular variation in chromosome copy number was true for tumors consisting of a mixture of (near) diploid and aneuploid subpopulations. These data indicate that DNA aneuploidy is associated with CIN but that (near) diploidy is not. Intratumoral regional DNA ploidy heterogeneity was conspicuous in 33 (92%) of 36 tumors with regions of DNA aneuploidy, and all aneuploid specimens showed great intercellular variation in chromosome copy number. Diploid regions were predominant in early stage cancers (intramucosal and submucosal cancers), and five of eight early cancers contained only diploid population. In contrast, all tumors without (near) diploid regions were advanced cancers. These observations suggest that CIN is a necessary prerequisite for developing intratumoral DNA ploidy heterogeneity with DNA aneuploidy.

Expression of p21(WAF1/CIP1) and p53 proteins in gastric carcinoma: its relationships with cell proliferation activity and prognosis.

The expressions of p21(WAF1/CIP1), p53 proteins, and Ki-67 antigen were investigated immunohistochemically in 190 primary gastric carcinomas. Of the 190 tumors, 40.5% positively expressed p21(WAF1/CIP1) and 42.1% positively expressed p53. The expression of p21(WAF1/CIP1) was significantly associated with clinicopathological factors including gender, tumor size, status of lymph node, and clinicopathological stage (P<0.05 for all), but p21(WAF1/CIP1) expression showed no clear correlation with Ki-67 labeling index. The mean Ki-67 labeling index was significantly higher in p53-positive cases than p53-negative cases (P<0.0001). However, among the clinicopathological factors examined, expression of p53 correlated only with age. Univariate and multivariate survival analyses revealed that clinicopathological stage (P<0.001) and expression status of p21(WAF1/CIP1) (P<0.05) were independent prognostic factors. Neither the expression status of p53 nor the Ki-67 labeling index, however, influenced the prognosis of patients with gastric cancer.

Diffuse cystic renal dysplasia: nonsyndromal familial case.

We report on a family in which three individuals, a male and two females were affected with nonsyndromal diffuse cystic dysplasia of the kidneys. The parents had no renal abnormality. The occurrence of diffuse cystic dysplasia in three sibs born to normal parents suggests autosomal recessive inheritance.

Comparative genomic hybridization analysis reveals 3q gain resulting in genetic alteration in 3q in advanced oral squamous cell carcinoma.

We analyzed DNA sequence copy number aberrations (DSCNAs) in 17 primary oral squamous cell carcinomas (OSCCs) by comparative genomic hybridization. DSCNAs were detected frequently at 3q25-qter (7/17), Xp21 (5/17), and Xq12-q23 and 8q23-q24 (4/17), and losses were detected frequently at 13q21-q22 (5/17), 3p21-pter, 4p15-pter and 17p13 (4/17), and 8p22-pter and 9p21-pter (3/17). Four tumors showed amplifications of seven loci: 3q11-qter, 3q13, 3q26, 7q21-q22, 8q23-qter, 9p22-pter, and 12p11. The total number of DSCNAs was significantly greater in stage III and stage IV tumors than in stage I and stage II tumors (P=.008). Furthermore, 3q gain was detected preferentially in stage III and stage IV tumors (6/8) rather than in stage I and stage II tumors (1/9, P=.013). In our study, all tumors with gain of 3q also contained one or more loss(es) in common regions. On the other hand, all tumors with gain of 9p did not contain 3q gains. These observations indicate that gain of 3q and accumulation of DSCNAs are strongly associated with tumor progression in OSCC. Furthermore, 3q gain and loss of one or more additional loci in common aberration regions appears to be a group of DSCNs associated with dominant genetic pathways of leading to advanced OSCCs.

DNA amplification and chromosomal translocations are accompanied by chromosomal instability: analysis of seven human colon cancer cell lines by comparative genomic hybridization and spectral karyotyping.

Genetic instability in human cancers is classified as chromosomal instability (CIN) or microsatellite instability (MIN). DNA amplification and translocations are observed frequently in various cancers. We used comparative genomic hybridization (CGH) and spectral karyotyping (SKY) to study seven human colon cancer cell lines and investigate the relations among genetic instability, DNA amplification, and chromosomal translocations. DNA amplification was found in five cell lines (COLO320DM, COLO201, WiDr, CoCM-1, and CACO-2), and all were aneuploid. In these five cell lines, segments of chromosomes were translocated to other chromosomes. In contrast, cell lines with MIN, DLD-1, and LoVo did not show DNA amplification. The LoVo cells with MIN were considered near diploid and contained translocations. These findings suggest that DNA amplification and chromosomal translocations are accompanied by CIN.

Cytogenetic analysis of esophageal squamous cell carcinoma cell lines by comparative genomic hybridization: relationship of cytogenetic aberrations to in vitro cell growth.

Cancer is characterized by autonomous growth of cells, and it is widely accepted that cell proliferation is primarily influenced by individual cell genetics. To elucidate the mechanisms of cancer cell proliferation, we studied differences in genetic aberrations for different type of tumors with different proliferation characteristics. We employed comparative genomic hybridization (CGH) to detect genetic aberrations in six cell lines of esophageal squamous cell carcinoma (ESCC). Three cell lines (YES-1, -2, and -3) grow in culture without fetal calf serum (group A), while others require serum to be maintained in vitro (group B). Both groups showed very similar cytogenetic aberrations: over-representations of 11q13 (6/6), 8q23-qter (5/6), Xq25-qter (5/6), 3q26-qter (4/6), 5p (4/6), 7p15-pter (4/6), 8q21.3-q22 (4/6), 17p (4/6), and 20q13 (4/6), and under-representations of 18q21-qter (6/6), 4q28-q33 (4/6), and 9p21 (4/6). Six amplification loci were mapped to chromosomal regions of 6q23 (1 case), 7p12 (2 cases), 9p21 (1 case), 11p11.2-12 (3 cases), 11q13 (2 cases), and 17p12 (2 cases). However, some differences were detected. DNA copy number increases at 7p12-p13, 11q14-q22, and 11q22-qter and under-representations of 4p, 8p, and 11p14-pter. In contrast, gains at 12p and 20p, and losses at 3p and 5q were detected only in group-B cell lines. These observations suggest that cytogenetic differences between the two groups may be linked to differences in cell growth characteristics in vitro, and that the genes in these chromosomal regions may play important roles in cell proliferation.

Examination of oncogene amplification by genomic DNA microarray in hepatocellular carcinomas: comparison with comparative genomic hybridization analysis.

To identify amplified oncogenes involved in hepatocellular carcinomas (HCC), we applied a genomic DNA microarray spotted with 57 oncogenes to 20 HCCs. Aberrations in DNA copy number also were analyzed by comparative genomic hybridization (CGH) using an aliquot of DNA samples. In 5 of 20 HCCs, only 6 oncogenes (CCND1, FGF3/FGF4, SAS/CDK4, TERC, MET, and MYC) were amplified, whereas in the remaining 15 tumors no oncogenes were amplified. A comparison of DNA microarray and conventional CGH analyses showed that, although 5 of 6 amplified oncogenes shown by microarray were located in chromosomal regions shown by CGH to have increased DNA copy numbers, not all genes located in such chromosomal regions were affected. One of the amplified oncogenes (SAS/CDK4) was found in a chromosomal region that was undetected by CGH. We, therefore, conclude that amplification of the oncogenes examined in this series is not directly implicated in hepatocellular carcinogenesis.

Amplification of 3q26 approximately qter correlates with tumor progression in head and neck squamous cell carcinomas.

Biological characteristics of malignant tumors including head and neck squamous cell carcinoma (HNSCC) are greatly affected by genetic alterations. However, the relationship between chromosomal aberrations and pathologic tumor stage in HNSCC has not been elucidated. In 32 patients, DNA sequence copy number aberrations (DSCNA) were surveyed by comparative genomic hybridization (CGH) combined with a microdissection method. The average number of DSCNA was 15.3 per tumor and increased with tumor stage (P<.05). DNA copy number gain was detected at 3q26 approximately qter in 29 tumors (91%), and 13 of these tumors displayed marked DNA amplification. Tumor stage was linked with this amplification (P<.05). The increase in DSCNA and amplification of 3q26 approximately qter are likely to be useful markers for estimating tumor progression in HNSCC.

Detection of 20q13 gain by dual-color FISH in breast cancers.

An increase in the 20q13 copy number has been reported in a wide range of solid tumors including breast cancers. However, the biological significance of this 20q13 gain has not yet been defined. We examined the 20q13 gain in breast cancer cells by dual-color FISH using two different DNA probes specific for the region of 20q13 and chromosome 20 centromere to investigate the relationship between 20q13 gain and the clinicopathological features of breast cancers. DNA measurement by LSC revealed DNA diploidy in 14 tumors and DNA aneuploidy in 24 tumors. Although the modal number of chromosome 20 was 2 in most of the tumors, the average fraction of cells with the modal chromosomal number was significantly different in the diploid and aneuploid tumors (p < 0.01). A gain in the 20q13 copy number was detected in 9 of 38 breast cancers, 2 of which showed a high-level gain. The gain in 20q13 was associated with negative expression for the progesterone receptor, but it was not linked to estrogen receptor expression. A 20q13 gain tended to be seen in DNA aneuploid and/or scirrhous carcinomas, but the increase in the 20q13 copy number did not affect either the nodal state or the disease stage in this series.

Chromosome 17 abnormalities in squamous cell carcinoma of the oral cavity, and its relationship with p53 and Bcl-2 expression.

Numerical aberrations of chromosome 17 were studied by fluorescence in situ hybridization (FISH) using pericentromere specific DNA probe in 27 oral squamous cell carcinomas (SCCs), and its relationship with p53 and Bcl-2 protein expression was investigated. Since cells with polysomy 17 are significantly increased in SCC (p = 0.0005), chromosome 17 abnormality seems to be correlated with carcinogenesis of oral SCC. Chromosome 17 abnormality varied from case to case, and the degree of the abnormality did not correlate with the stage of the disease, the histological differentiation of SCC, or relapse of the disease and lymph node metastasis. However, there was a correlation between polysomy 17 and p53 immunoreactivity (p = 0.0228). p53 immunoreactivity showed a correlation with relapse of the disease (p = 0.0441). An inverse relationship was found between p53 and Bcl-2 immunoreactivity (p = 0.0421). In conclusion, we suggest that polysomy 17 is closely related carcinogenesis of oral SCC, and with p53 overexpression. It is assumed that polysomy 17 correlates with the mutation of p53 which results in an accumulation of aberrant p53 protein, and that its activation causes an increase of p53 protein.

New molecular tumor markers for endometrial cancer.

Although the International Federation of Gynecology and Obstetrics officially changed the classification system of endometrial cancer from a clinically staged to a surgically staged disease in 1988, optimal management of patients with endometrial cancer is still controversial. Gynecologists happen to experience that patients with tumors that are identical in grade and stage often have significantly different clinical outcomes or responses to therapy. In order to identify an objective biological factor correlating with tumor aggressiveness, many tumor markers have been investigated. So far, CA125 is one of the most reliable tumor marker for adenocarcinoma of the uterus and frequently used in a clinical setting. Recently, with the advent of molecular biological techniques, many genes and regions of the genome related to endometrial cancer have been identified. We undertook a genome-wide screening to detect genetic changes by comparative genomic hybridization (CGH) in primary endometrioid cancers, since CGH analysis provides comprehensive information concerning relative chromosomal losses and gains in tumors by a single hybridization. In this paper, the usefulness of serum tumor markers and the new promising molecular tumor markers for endometrial cancer are discussed.

Congenital cystic adenomatoid malformation of the lung with an esophageal cyst: report of a case.

The authors report the case of a 7-month-old girl found to have both congenital cystic adenomatoid malformation of the lung (CCAM) and esophageal cyst. She suffered repeated episodes of pneumonia and exhibited signs of respiratory distress on admission to our hospital. Chest radiography and magnetic resonance imaging (MRI) showed 2 different kinds of cystic lesions. Resection of the lower lobe of the right lung and excision of the posterior mediastinal cyst were performed. Histologic examination showed Stocker type I CCAM and esophageal cyst. Coexistence of both CCAM and esophageal cyst is extremely rare. The authors speculate that the pathologies of this case originated from a regional disturbance of common embryologic origin during 2 different phases of lung-bud foregut malformations.

[Simultaneous measurement of kinetochore protein and DNA by flow cytometry].

We were successful in obtaining simultaneous measurements of kinetochore protein and DNA with flow cytometry using HeLa cells and KM cells; the latter originated with a squamous cell carcinoma of the oral cavity. Kinetochore protein was stained with the serum obtained from a patient with CREST syndrome and FITC-conjugated goat antihuman IgG antibody; the DNA was stained with propidium iodide. The FITC fluorescence of cells with a large DNA index was stronger than in cells with a small DNA index. The FITC intensity increased from G1 phase to G2/M phase. The rate of FITC intensity increase representing the kinetochore protein amount was greater in the early and late S phases than in the other phases. The results suggest that kinetochore protein increases in the interphase as the cell cycle unfolds.

[A simple method for flow cytometric measurement of Ki-67 reactive antigen on small solid tissues].

Monoclonal antibody Ki-67 is a useful and easy means for evaluating cell proliferative activity and malignancy of tumor cells. However, almost all studies for expression of Ki-67 reactive antigen on solid tumors have been performed only by microscopic observation of immunostained sections, and not by flow cytometry. The main reason for this is the difficulty associated with preparation of numerous suspended cells so as to be analyzable with FCM from small solid tissues. In this study, a simple and easy method for flow cytometric measurement of Ki-67 reactive antigen on small solid tissues was shown. The important steps in this method are cell dispersion with Triton X-100 and washless staining.