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Resume Objectifs Evaluer la faisabilite et l’interet du Doppler cerebral fœtal en cours de travail et son lien avec le pH au scalp pour predire l’asphyxie fœtale per partum. Methodes Etude prospective unicentrique en maternite universitaire de type III incluant des patientes en cours de travail. Patientes sans facteur de risque, presentant des grossesses de deroulement normal au-dela de 37 semaines d’amenorrhee. Dans chaque cas, une echographie avec Doppler cerebral a ete realisee de maniere concomitante a la realisation d’un pH au scalp. Ont ete relevees les caracteristiques maternelles et fœtales ainsi que la dilatation cervicale, l’analyse du rythme cardiaque fœtal et la variete de presentation. Resultats Sur 49 patientes incluses durant une periode de 4 mois en salle de travail, 7 echecs de realisation du Doppler (11 %) ont ete recenses survenant, le plus souvent, a dilatation complete (p = 0,007, OR = 14,1 [1,483 ; 709,1275]). Les autres facteurs : âge, indice de masse corporelle, parite, uterus cicatriciel n’ont pas ete retrouves comme significativement lies a un echec de realisation. Aucune correlation significative entre Doppler cerebral et pH au scalp (r = 0,15) ni pH a la naissance (r = 0,13) n’a ete mise en evidence. Aucun seuil de Doppler cerebral predictif de l’asphyxie n’est mis en evidence. Conclusions Le Doppler cerebral est realisable en cours de travail mais n’est donc pas un bon examen de seconde ligne pour l’evaluation de l’acidose fœtale en cours de travail.

Introduction et but de l’etude Le surpoids et l’obesite sont de reels problemes de sante publique, par leur frequence et leurs consequences. Le medecin generaliste est en premiere ligne pour depister et agir precocement sur les determinants comportementaux de l’exces ponderal, et est demandeur de formation dans ce domaine. L’objectif de cette boite a outils est de lui fournir des « cles » d’aide a la prise en charge de ses patients, basees sur des approches therapeutiques non « restrictives » et non stigmatisantes. Elles devraient permettre a tout medecin interesse, d’acceder a ces nouvelles approches therapeutiques, et l’aider a structurer sa prise en charge. Materiel et methodes Une revue de la litterature et des outils disponibles sur internet, a ete realisee fin 2018 et n’a pas permis de retrouver de procedure dediee a la prise en charge medicale de premier recours pour l’identification et la prise en charge des differentes dysregulations du comportement alimentaire favorisant l’exces ponderal. Ce travail innovant a ete realise par l’equipe du Centre Specialise de l’Obesite de Champagne-Ardenne. Resultats et analyse statistique Cette « Boite a Outils » est accessible librement via le site internet de « La Maison de la Nutrition » : https://www.maison-nutrition.fr/professionnels/boite-a-outils-professionnel-de-sante/Y sont presentes sous une structuration identique, les differents elements a rechercher et a caracteriser sur le plan du comportement alimentaire (tachyphagie–hyperphagie prandiale–prises extra-prandiales), face a un exces ponderal. Ces elements sont explicites et suivent le fil d’une consultation pour en faciliter l’utilisation. Des fiches recapitulatives pour le medecin et des fiches educatives a destination du patient (« take home messages ») sont disponibles pour chaque dimension exploree. Une evaluation de cet outil est disponible sur le site. Cet outil est en cours de presentation aupres des acteurs locaux, via des formations medicales continues. Conclusion La « Boite a Outils » Surpoids/Obesite a destination des medecins generalistes a pour but d’ameliorer leurs pratiques de soins, en leur proposant une alternative aux regimes restrictifs (comme le recommande l’ANSES depuis 2010).

To evaluate feasibility and interest of fetal cerebral Doppler during labor and the link with fetal pH to predict perinatal fetal asphyxia.Our prospective study in a university perinatal center, included patients during labor. There were no risk factors during pregnancy and patients were included after 37 weeks of pregnancy. For each patient an ultrasound with cerebral Doppler was done concomitant to a fetal scalp blood sample. We collected maternal and fetal characteristics as well as cervix dilatation, fetal heart rate analysis and fetal presentation.Among 49 patients included over a period of 4 months, cerebral Doppler failed in 7 cases (11%). Majority of failure occurred at 10cm of dilatation (P=0.007, OR=14.1 [1.483; 709.1275]). Others factors like: maternal age, body mass index, parity, history of C-Section were not associated with higher rate of failure. We did not found either significant correlation between cerebral fetal Doppler and pH on fetal scalp blood sample (r=0.15) nor pH at cord blood sample (r=0.13). No threshold of cerebral Doppler is significant for fetal asphyxia prediction.Fetal cerebral Doppler is feasible during labor with a low rate of failure but not a good exam to predict fetal acidosis and asphyxia.

In order to assess the effect of in vitro models on the expression of key genes known to be implicated in the development or progression of cancer, we quantified by real-time quantitative PCR the expression of 28 key genes in three bladder cancer tissue specimens and in their derived cell lines, studied either as one-dimensional single cell suspensions, two-dimensional monolayers or three-dimensional spheroids. Global analysis of gene expression profiles showed that in vitro models had a dramatic impact upon gene expression. Remarkably, quantitative differences in gene expression of 2–63-fold were observed in 24 out of 28 genes among the cell models. In addition, we observed that the in vitro model which most closely mimicked in vivo mRNA phenotype varied with both the gene and the patient. These results provide evidence that mRNA expression databases based on cancer cell lines, which are studied to provide a rationale for selection of therapy on the basis of molecular characteristics of a patient's tumour, must be carefully interpreted. British Journal of Cancer (2002) 86, 1283–1289. DOI: 10.1038/sj/bjc/6600239 www.bjcancer.com © 2002 Cancer Research UK

We have isolated a new -ene, named BAC, which is the human equivalent of the murine H19 gene and is highly expressed in most fetal tissues and in a variety of fresh tumors. BAC was analyzed in 130 untreated invasive carcinomas of different types. The frequency of BAC-expressing cancers as well as the level of expression greatly varied among the different types of cancer and within the same type of cancer. For example, the 2.3 kb BAC transcript band was detected in 94% of breast adenocarcinomas and in only 35% of epidermoid lung carcinomas with differences of 100-fold in the level of expression between tumor specimens. The majority of tumor tissues displayed BAC expression while their normal counterpart did not with the exception of normal breast tissues which contained low but significant level of BAC transcript. It is possible that BAC expression was influenced by the presence of gene deletions in tumors. Indeed, this gene is located in chromosome 11p15, a region in which deletions have frequently been observed in human cancers. Therefore, the variable levels of expression could have a biological significance and be used as a marker of tumor progression.

Purpose The aim was to investigate whether germline polymorphisms within candidate genes known or suspected to be involved in fluorouracil (FU), oxaliplatin, and irinotecan pathways were associated with toxicity and clinical outcome in patients with metastatic colorectal cancer (mCRC). Patients and Methods Blood samples from 349 patients included in the Fédération Francophone de Cancérologie Digestive 2000-05 randomized trial, which compared FU plus leucovorin (LV5FU2) followed by FU, leucovorin, and oxaliplatin (FOLFOX) followed by FU, leucovorin, and irinotecan (FOLFIRI; sequential arm) with FOLFOX followed by FOLFIRI (combination arm) in terms of progression-free survival (PFS) and overall survival, were collected. Twenty polymorphisms within the DPD, TS, MTHFR, ERCC1, ERCC2, GSTP1, GSTM1, GSTT1, and UGT1A1 genes were genotyped. Results The ERCC2-K751QC allele was independently associated with an increased risk of FOLFOX-induced grade 3 or 4 hematologic toxicity (P = .01). In the sequential arm, TS-5′UTR3RG and GSTT1 alleles were independently associated with response to LV5FU2 (P = .009) and FOLFOX (P = .01), respectively. The effect of oxaliplatin on tumor response increased with the number of MTHFR-1298C alleles (test for trend, P = .008). The PFS benefit from first-line FOLFOX was restricted to patients with 2R/2R (hazard ratio [HR] = 0.39; 95% CI, 0.23 to 0.68) or 2R/3R (HR = 0.59; 95% CI, 0.42 to 0.82) TS-5′UTR genotypes, respectively. Conversely, patients with the TS-5′UTR 3R/3R genotype did not seem to benefit from the adjunction of oxaliplatin (HR = 0.96; 95% CI, 0.66 to 1.40; trend between the three HRs, P = .006). Conclusion A pharmacogenetic approach may be a useful strategy for personalizing and optimizing chemotherapy in mCRC patients and deserves confirmation in additional prospective studies.

The aim of the present study was to analyse in invasive carcinomas of the uterine cervix, the anionic glutathione S transferase (GST pi) gene, possibly implicated in the drug resistance of human cancers. Total RNA preparations obtained from invasive cervical cancers (106 specimens), carcinomas in situ (CIS) (three specimens) and normal cervical epitheliums (24 specimens) were analysed by Northern and slot blot hybridisation. A 0.7 kb GST pi transcript band was detected in all the cervical specimens. GST pi mRNA levels were lower in normal cervix (mean: 0.7 +/- 0.1 arbitrary units) than in invasive carcinomas (mean: 2.5 +/- 1.5 units) (Student test P less than 10(-4)). However no significant difference was observed between invasive cancers of advanced stages (III and IV) and those of early stages (I and II). The presence of human papillomavirus in cancers and in normal cervices did not influence significantly the GST pi mRNA level. Neither amplification nor gross rearrangement of GST pi gene could be observed after Southern blot analysis of genomic DNA. In conclusion, our data indicate that the presence of high levels of GST pi transcripts in invasive cancers may be a consequence of the multiple biochemical changes which accompany cervical carcinogenesis. Images Figure 1 Figure 2

International audience; Mutations in two genes encoding cell cycle regulatory proteins have been shown to cause familial cutaneous malignant melanoma (CMM). About 20% of melanoma-prone families bear a point mutation in the CDKN2A locus at 9p21, which encodes two unrelated proteins, p16(INK4a) and p14(ARF). Rare mutations in CDK4 have also been linked to the disease. Although the CDKN2A gene has been shown to be the major melanoma predisposing gene, there remains a significant proportion of melanoma kindreds linked to 9p21 in which germline mutations of CDKN2A have not been identified through direct exon sequencing. The purpose of this study was to assess the contribution of large rearrangements in CDKN2A to the disease in melanoma-prone families using multiplex ligation-dependent probe amplification. We examined 214 patients from independent pedigrees with at least two CMM cases. All had been tested for CDKN2A and CDK4 point mutation, and 47 were found positive. Among the remaining 167 negative patients, one carried a novel genomic deletion of CDKN2A exon 2. Overall, genomic deletions represented 2.1% of total mutations in this series (1 of 48), confirming that they explain a very small proportion of CMM susceptibility. In addition, we excluded a new gene on 9p21, KLHL9, as being a major CMM gene.

The c-Ha-ras gene was analysed by Southern blot hybridisation in 67 specimens of lymph node metastases and in 25 specimens of primary tumours obtained from 85 untreated patients with head and neck squamous cell carcinoma. The loss of one c-Ha-ras allele was observed in 10/46 (22%) tumours from heterozygous patients for this locus. Different genes, located as the c-Ha-ras gene on the short arm of chromosome 11, were also found to be deleted suggesting that the deletion of other genes could play a role in aggressiveness of head and neck carcinomas. Using polymerase chain reaction, mutation at codon 12 was detected in only 2/54 (3.8%) tumours but no mutation involving codon 61 was found. Neither gene amplification nor gene rearrangement could be observed. Total RNA was prepared from 79 of these tumour specimens and analysed by Northern and slot blot hybridisation. A 1.2 kb c-Ha-ras transcript band was detected in all the RNA preparations. Relatively high c-Ha-ras transcript levels were found in 18% of lymph node metastases and in 21% of primary tumours, indicating no significant differences between these cancers. Moreover, the c-Ha-ras mRNA levels were not significantly greater in the primary tumours than in the normal mucosae in 10/12 cases for which both tissues were analysed. These data indicate that c-Ha-ras gene does not seem to be strongly involved in head and neck carcinomas at that advanced stage of the disease, as this was previously reported for earlier clinical stages. Images Figure 1 Figure 3 Figure 4 Figure 6 Figure 7 Figure 8

BRCA1 and BRCA2 germline mutations, mainly point mutations and other small alterations, are responsible for most hereditary cases of breast-ovarian cancer. However, the observed frequency of BRCA1 alterations is lower than that predicted by linkage analysis. Several large BRCA1 rearrangements have been identified with a variety of technical approaches in some families. We have developed a gene dosage assay based on real-time quantitative PCR and used it to extensively analyze 91 French families of breast-ovarian cancer in which no BRCA1 or BRCA2 point mutations was identified. This gene dosage method calculates the copy number of each BRCA1 exon to readily detect one, two, and three or more copies of BRCA1 target exons. In the series of 91 families at high risk of carrying BRCA1 mutations, we detected seven large rearrangements of the BRCA1 gene by using this real-time PCR approach. This simple, rapid, and semiautomated real-time quantitative polymerase chain reaction (PCR) assay is a promising alternative technique to Southern blot, bar code analysis on combed DNA, quantitative multiplex PCR of short fluorescent fragments, and cDNA length analysis for the detection of large rearrangements. Therefore, this technique should be considered as a powerful diagnostic method for breast/ovarian cancer susceptibility in clinical and research genetic surveys.

The p53 gene homologue, p73, is located on the 1p36-3 locus, which is frequently deleted in human neuroblastoma (NB). A survey of 61 NB showed that among 33% of informative cases, p73 loss of heterozygosity (LOH) occurred in 7 of 20 (35%).LOH pattern of vicinal markers suggested that the p73 gene could not be considered as the candidate NB suppressor gene. Moreover, comparative measurements of allelic expression in tumors and corresponding patient lymphocytes indicate that pure biallelism is much more frequent in lymphocytes than in tumors (71% vs 30%, P= 0.05), which suggests that disequilibrated allelic expression is associated with NB disease.Therefore, in the p73 LOH NBs, the p73 gene could be altered in the maintained allele not by mutations [Ishimiya et al.: Med Pediatr Oncol, this issue], but rather by an abnormal transcription.

The paper deals with the recent improvements introduced in the most usual method applied in the Institut Gustave Roussy radiotherapy department for obtaining the anatomical data of patients treated for head and neck tumors. For each of these patients, five to seven transverses slices and a lateral radiographic film are taken from a Mecaserto simulator-CT. The anatomical representation of the patient sagittal plane is carried out from the digitalisation of the radiographic film on a Vidar Vxr-12 Plus film scanner and integrated into the Dosigray dose calculation programme in order to be used as a support for the laying out of the dose distribution in reference to the treatment. The sagittal anatomical representation obtained from the radiographic film digitalisation is compared with the one resulting from the interpolation between a limited number of irregularly-spaced transverse slices taken on the simulator-CT. The method using the simulator-scanner transverse slices and the radiographic film digitalisation represents an interesting alternative for obtaining an anatomy simulation representative of the patient in hospitals where a scanner is not available full-time for the needs of the radiotherapy process.

Our aim was to compare the prognostic value of c-erbB-2 gene amplification analyzed by Southern blot with that of protein (p185) over-expression measured by immunohistochemistry in 172 patients with operable breast cancer (BC). Amplification and p185 over-expression were found in 31 (18%) and 51 (30%) BCs, respectively. All but 1 of the tumors showed both amplification and over-expression, while 21 (12%) tumors displayed over-expression without amplification. The risk of death associated with c-erbB-2 gene amplification and p185 over-expression was evaluated by multivariate analysis, taking into account tumor size, histoprognostic grade, hormone receptors and axillary node status. During a mean follow-up of 9.5 (+/-2) years, node involvement (p0.001), c-erbB-2 gene amplification (p = 0.02) and negative hormone receptors (p = 0.02) were found to be independent prognostic indicators of the risk of death. Over-expression of p185 with no amplification was not correlated with this risk. When the risk of death associated with c-erbB-2 amplification was studied according to chemo- and hormone therapy, no significant difference was observed between subgroups of subjects. Amplification was also associated (p = 0.02) with the risk of multifocal distant metastases (i.e., metastases detected concomitantly in at least 2 sites) and, thus, with BC aggressiveness. These data show the importance of c-erbB-2 gene amplification in predicting the long-term outcome of patients and in selecting eligible patients for c-erbB-2-targeted therapies.

Neuroblastoma is a very common solid tumor which arises in childhood and shows an extreme heterogeneity at the clinical, histological and genetic levels. Besides age and stage, N-myc amplification and 1p deletion are prognostic factors of the disease: in Europe, these genetic markers are used to conduct therapy. In France, N-myc amplification is a factor of bad prognosis which leads, in all forms of the disease including localised forms and metastatic forms of children aged of less than 1 year, to a myeloablative treatment with autologous hematopoietic stem cells transplantation. By contrast, N-myc amplification has no impact on the survival of children aged of more than 1 year with a poor prognosis (30% overall survival, 5 years) but this genetic abnormality is taken into account to treat primary tumor of these patients. In an attempt to find out prognostic factors of these aggressive forms of the disease, various pathways (apoptosis, differentiation angiogenesis, detoxication, immune response) have been recently surveyed, but studies have been carried out on a limited number of genes. Moreover, experimental models of human metastatic neuroblastoma have been obtained in which variations of genes transcript levels involved in these pathways, are observed. The current break-through of cDNA microarrays allows to develop a dynamic transcriptomic scanning of these models as well as of tumors and bone marrows from patients upon conventional chemotherapy. This technology will enable: i) to define molecular entities of the metastatic disease; ii) to apply adapted treatment; iii) to develop new therapeutic strategies.

Disseminated neuroblastoma frequently show a very poor prognosis. N-myc gene amplification, 1p deletion and lack of CD44 gene expression, are all genetic factors associated with the disease's dissemination. Human neuroblastoma xenografts in nude mice has permitted to characterize, in disseminated neuroblasts, oncogenes overexpression, inactivation of tumor suppressor genes as well as detoxifying genes activation which contributes to increase cellular resistance to chemotherapy. These genetic abnormalities permit to propose a nosology of this very aggressive pediatric solid tumor. Hopefully, this genetic classification could be of great value for new therapeutic approaches.

Mutation of the p53 tumor suppressor gene is the most commonly observed gene alteration in human cancers. In order to identify new prognostic factors and tumor aggressiveness in squamous cell head and neck carcinomas, we analyzed 50 node metastases and 28 primary tumors including 13 matched specimens for p53 alterations. Mutations were found in 54 (69%) tumors, 76% of which were missense, 9% were nonsense and 15% were microdeletions or microinsertions. Twenty-five mutations were transitions mostly G--A (40%) and 20 were transversions mostly G--T (25%) thus confirming the role of tobacco carcinogens in the induction of these mutations. For eight patients mutations were observed in matched primary tumors and metastases, indicating clonal dissemination of tumor cells in most of these carcinomas. Furthermore the incidence of mutations was not different in primary tumors and node metastases indicating that this gene alteration was not related to the metastatic dissemination. No correlation was found between mutation and clinical parameters, the 8-year survival rates were not different (log rank test: P = 0.49) in patients with and without mutation. There was a good correlation between p53 mutation and protein overexpression (Fisher's exact test: P10(-4). Interestingly, immunostaining was also observed in basal cells from normal mucosa and in early lesions adjacent to the primary tumor in 11/15 specimens irrespective of the presence of mutation in the corresponding tumors. p53 protein overexpression may therefore constitute a biomarker for early stages of carcinogenesis of the head and neck epithelium.

The few imprinted genes characterized so far include the insulin-like growth factor-2 gene (IGF2) coding for a foetal growth factor and the H19 gene whose normal function is unknown but which is likely to act as an RNA with an antitumour effect. IGF2 is expressed by the paternal allele and H19 by the maternal allele. This reciprocal expression is quite interesting because both H19 and IGF2 genes are located close to each other on chromosome 11p15.5 in a region subject to loss of heterozygosity (LOH). Moreover, loss of imprinting (LOI) or biallelic expression has been proposed as an epigenetic mechanism for tumorigenesis in a variety of human cancers including Wilms' tumour. In this study we report the LOH, LOI and methylation status of H19 and IGF2 genes in 29 invasive cervical carcinomas of different clinical stages. Fourteen (48%) and 13 (45%) tumours were heterozygous for H19 and IGF2 respectively. LOH for H19 and IGF2 genes were found in 2 of 14 (14%) and 3 of 13 (23%) informative tumours, respectively. LOI of H19 and IGF2 was detected in 2 of 12 (17%) and 5 of 10 (50%) tumours with no LOH, respectively. More interestingly, monoallelic expression of the otherwise silent H19 allele (allele switch) was observed in 2 of 12 (17%) tumours and biallelic expression of IGF2 was detected in one specimen of normal cervix adjacent to the tumour. The expressing H19 allele, and to a lower degree also the silent allele, were hypomethylated in tumours suggesting that demethylation of both H19 alleles may be associated with an early step of imprinting alteration. In cervical cancer H19 and IGF2 expressions could be independently regulated. In conclusion, our data suggest that H19 and IGF2 genes, via deletions and/or abnormal imprinting, could play a crucial role in a large proportion (58%) of cervical cancers where they may be associated with disease progression.

We have analysed 78 head and neck carcinomas (50 node metastases and 28 primary tumors including 13 matched specimens) in 65 patients for p53 alterations. Mutations were found in 54 (69%) tumors. Of the 53 mutations within exons, 40 (76%) were missense, five (9%) nonsense and eight (15%) microdeletions or microinsertions. Twenty-five (47%) mutations were transitions mostly G--A (40%) and 20 (38%) were transversions, mostly G--T (25%), thus confirming the role of tobacco carcinogens in the induction of these mutations. The incidence of mutations was not different in primary tumors (68%) and node metastases (70%) indicating that this gene alteration was not related to the metastatic dissemination. For eight patients, mutations were observed in matched primary tumors and metastases, indicating clonal dissemination of tumor cells in most of these carcinomas. There was a good correlation between mutations and protein overexpression (Fisher's exact test P10(-4). Immunostaining was also observed in basal cells from normal epithelium and in early lesions adjacent to the primary tumor in 11/15 (73%) specimens irrespective of the presence of mutation in the corresponding tumors. These data confirm that p53 overexpression is an early event in the multistep process of epithelial cell carcinogenesis. Loss of heterozygosity for the TP53 locus was detected in 54% of tumors but no association was found with mutation (Fisher's exact test P = 0.14). No mdm-2 amplification was detected in any tumors. No correlation was found between mutation and clinical parameters, the 5-year survival rates were not different (log rank test P = 0.39) in patients with and without mutation. In conclusion, we have shown that p53 gene mutations and deletions and protein overexpression are frequent in the most aggressive head and neck carcinomas but are not associated with disease progression. The presence of protein in normal mucosa and in non-invasive lesions may constitute a biomarker for early stages of carcinogenesis.

9000 Background: Integrative studies correlating DNA changes and expression data are powerful to identify new genetic defects involved in tumor progression. In order to identify new pathways involved in melanoma progression and to better understand the gender effect on melanoma prognosis, we correlated micro-array comparative genomic hybridization (aCGH) with expression levels of several genes in frozen melanoma samples. Methods: 48 primary melanomas from 32 females and 16 males with a median follow-up of 4 years (range: 0.6–15 years), and for which DNA and RNA were co-extracted from the same frozen slices, were analysed using long oligo 244K aCGH slides (Agilent Technologies; Pao Alto). Each sample was hybridized in single versus a standard pool of DNA with matched sex. Expression data and copy number modifications were correlated and adjusted for the False Discovery Rate. The active or inactive X chromosome (chr) status was characterized by RNA FISH with a DNA probe detecting Xist RNA. The relative abun...

The biological behaviour of invasive carcinoma of the uterine cervix is not always predictable. It is therefore important to establish new biological markers which could be useful in determining a more reliable prognosis. We have analyzed the c-Ha-ras and c-myc proto-oncogenes in a large series (154 cases) of cervical cancers at various clinical stages. Alterations of c-Ha-ras (deletion, mutation) and c-myc (amplification) were frequently observed in cervical cancers and were shown to be associated with tumor progression. Furthermore, c-myc overexpression, when detected in early cervical cancers, provides a means of identifying patients at high risk of early recurrence.

2508 Background: The FFCD 2000–05 randomized trial compared simplified LV5FU2 followed by FOLFOX6 (arm 1) to FOLFOX6 followed by FOLFIRI (arm 2) in the treatment of metastatic colorectal cancer. The aim was predicting the toxicity profile of oxaliplatin after the first line treatment using pharmacogenetic data. Methods: Patients (pts) with available blood samples were compared to the other pts for clinical prognostic factors (chi2 test). A logistic model was computed to test the association between polymorphisms and toxicity in each arm. An interaction test was used to assess a differential effect according to treatment (predictive effect), in order to identify a predictive effect of oxaliplatin. Grade 3–4 hematological and non-hematological toxicities (H-tox and NH-tox) at 4 months and grade 2–4 neurological at 6 months were the endpoints of the study. Thirteen genetic variants in 10 candidate genes were selected for pharmacogenetic analysis: ERCC1_04 (rs3212961), ERCC1_05 (rs11615), ERCC1_06 (rs3212948), ERCC1_24 (rs3212955), ERCC2_02 (rs1799793), ERCC2_03 (rs13181), ERCC2_06 ( rs238406 ), ERCC2_09 (rs1799787), GSTM1 (null/present), GSTT1 (null/present), TS (TSER, Ins/del6bp) and UGT1A1 (rs8175347). Genotyping was performed using Taqman probes, QMPSF and fragment analysis. Results: 327 pts (156/171) out of 410 were included (61 had no blood samples, 16 had less than 2 cycles, 3 had incomplete data on toxicity, 3 had insufficient DNA). No difference was found between included and excluded pts in the analysis for gender, age, OMS, number of metastatic organs and adjuvant chemotherapy. Pts received similar 5FU doses in both arms. Number of patients with at least one toxicity in arms 1/2 were as followed: 5/54 grade 3–4 H-tox, 28/47 grade 3–4 NH-tox, and 0/103 grade 2–4 neurological. The genotype CC of ERCC2_02 correlated with higher NH-tox at 4 months in arm 2 (p=0.0008, OR=0.31, 95%CI=[0.15–0.62] versus p=0.87, OR=0.93, CI=[0.39–2.21] in arm 1) compared to genotypes CT and TT, with borderline interaction (p=0.05). Conclusions: These preliminary results on early toxicity in first-line are in favour of an effect of ERCC2_02 on NH-tox of FOLFOX6 and a predictive effect on NH-tox of oxaliplatin. [Table: see text]

The c-Ha-ras-1 locus was analysed in cervical cancers and shown to exhibit the loss of one allele in 36% of heterozygous tumours and a mutation at codon 12 in 24% of tumours at advanced stages. 40% of tumours with mutation contained also a deletion. A c-myc gene activation was found in 100% and 70% of tumours containing mutation and deletion respectively. This suggests that the two proto-oncogenes cooperate for the progression of cervical cancers. Furthermore as more than 90% of these tumours contained also human papillomavirus sequences, our data strongly suggest that multiple genetic events are involved in the genesis and progression of most cervical cancers.

Invasive squamous cell carcinomas of the uterine cervix from 12 untreated patients were examined for the presence of human papillomavirus (HPV) genomes and for the state of the oncogenes c-myc and c-Ha-ras. Blot hybridization experiments have demonstrated the presence of the genome of HPV type 16 (HPV 16) in six tumors and that of the genomes of HPV types weakly related to HPV 16 or HPV 18 in five others. In the nine tumors corresponding to advanced stages of the disease (stages 3 and 4) there was a 3-30 fold amplification of c-myc and/or c-Ha-ras. A concomitant amplification of both oncogenes was found in eight cancers. In only one of the three tumors confined to the cervix (stage 1), the oncogene c-Ha-ras was weakly amplified. Neither HPV DNA sequences, nor oncogene amplification were detected in the leukocytes of five patients. Thus, it seems likely that specific HPV types play a role in the development of carcinomas of the uterine cervix, and that cellular oncogenes, activated through an amplification process, are involved in at least some steps of tumor progression.

The cleavage of the kDNA minicircles of Trypanosoma equiperdum by the restriction endonucleases Hinf I, Bgl II, Mbo I, Tag I and Mbo II revealed that this kDNA is homogeneous in base sequence. This is in contrast with the kDNA of minicircles of the other species of trypanosomes so far studied. The 10 cleavage sites, obtained with these endonucleases, were ordered and a restriction cleavage map of the minicircles was thus drawn.

Tumor specimens from 116 untreated patients with primary breast carcinoma at different clinical stages were analyzed for the structure and/or the expression of c-myc and c-erbB-2/neu proto-oncogenes. An amplification of the c-myc proto-oncogene (3 to greater than 50 fold) was detected only in 6% of carcinomas, with no evidence of locus rearrangement. High c-myc RNA levels detected in 45% of tumors were found significantly (p less than 0.01) correlated with lymph node involvement. Amplification (3 to greater than 30 fold) of the c-erbB-2/neu gene was observed in 20% of cancers. A 5 kb c-erbB-2/neu gene transcript was detected in the 103 cancer specimens analyzed. High levels of transcripts were observed in 36% of tumors. Overexpression did not depend only on amplification since found in 14 tumor samples with a single gene copy. The gene amplification and overexpression were found significantly associated with cancers of poor prognosis. Moreover our data show that both proto-oncogenes are overexpressed only in 12.5% of tumor samples and suggest that each gene might play a different role in tumor progression.

International audience; So far, no common environmental and/or phenotypic factor has been associated with melanoma and renal cell carcinoma (RCC). The known risk factors for melanoma include sun exposure, pigmentation and nevus phenotypes; risk factors associated with RCC include smoking, obesity and hypertension. A recent study of coexisting melanoma and RCC in the same patients supports a genetic predisposition underlying the association between these two cancers. The microphthalmia-associated transcription factor (MITF) has been proposed to act as a melanoma oncogene; it also stimulates the transcription of hypoxia inducible factor (HIF1A), the pathway of which is targeted by kidney cancer susceptibility genes. We therefore proposed that MITF might have a role in conferring a genetic predisposition to co-occurring melanoma and RCC. Here we identify a germline missense substitution in MITF (Mi-E318K) that occurred at a significantly higher frequency in genetically enriched patients affected with melanoma, RCC or both cancers, when compared with controls. Overall, Mi-E318K carriers had a higher than fivefold increased risk of developing melanoma, RCC or both cancers. Codon 318 is located in a small-ubiquitin-like modifier (SUMO) consensus site (ΨKXE) and Mi-E318K severely impaired SUMOylation of MITF. Mi-E318K enhanced MITF protein binding to the HIF1A promoter and increased its transcriptional activity compared to wild-type MITF. Further, we observed a global increase in Mi-E318K-occupied loci. In an RCC cell line, gene expression profiling identified a Mi-E318K signature related to cell growth, proliferation and inflammation. Lastly, the mutant protein enhanced melanocytic and renal cell clonogenicity, migration and invasion, consistent with a gain-of-function role in tumorigenesis. Our data provide insights into the link between SUMOylation, transcription and cancer.