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Context and Aims: Breast cancer prognosis and predictive biomarkers development would allow sparing some patients from chemotherapy or identifying patients for whom chemotherapy would be indicated. In this context, in 2009, the French National Cancer Institute, a National Health and Science Agency dedicated to cancer, in collaboration with the French society of senology and breast pathology (SFSPM) published a report on the assessment of the prognostic and the predictive clinical validity of tissular biomarkers, uPA/PAI-1, Oncotype DX™ and MammaPrint(®), in breast cancer management. They concluded that only the uPA/PAI-1 prognosis value reached the highest level of evidence (LOE I according to Hayes 1998 classification). In 2012, it was decided to update this report since new data have emerged and because information disparities among clinicians have been identified. This article aims to present the main conclusions together with the levels of evidence associated with those conclusions., Methods: The updating process was based on literature published since 2009 appraisal and on multidisciplinary and independent experts' opinion. The levels of evidence (LOE) used are those of the classification defined by Simon in 2009 (updated Hayes 1998 classification): LOE IA and LOE IB: high level of evidence; LOE IIB and LOE IIC: intermediate level of evidence; LOE IIIC and LOE IV-VD: low level of evidence., Conclusions: Among patients without lymph-node involvement, uPA/PAI-1, invasion process biomarkers, reach the highest level of evidence for 10 years recurrence free survival prognosis (LOE IA according to Simon). The predictive value to anthracyclins chemotherapy remains to be confirmed. Oncotype DX™ and MammaPrint(®) prognosis and predictive value do not reach the LOE I level. This updating' process confirms the 2009 levels of evidence for all the three biomarkers prognosis value. Besides, concerning Oncotype DX™ and MammaPrint(®), new data do not allow to conclude neither to their complementary clinical information to other clinicopathological existing biomarkers nor to a favorable cost-efficiency ratio in therapeutic decision making and this because of the methodological weakness and uncertainty that are identified in the selected studies. Practically, beyond the prognosis and predictive biomarkers validity, the clinical utility of a new biomarker for chemotherapy indication depends on its clinical added information with regard to validated biomarkers (HR, HER2 and Ki67) and to clinicopathological parameters. Since they are the sole validated biomarkers of the invasion process, uPA/PAI-1 could complete clinical information of other clinicopathological factors and consequently could confer an added clinical value. However, data concerning the impact of this information on chemotherapy clinical indication are lacking., (Copyright © 2014. Published by Elsevier Masson SAS.)

As part of a study in the North of France for screening pelvic tumours with plasma proteomic analysis, we included 82 women with hereditary risk of ovarian cancer. We report here the consequences of organized screening with usual tests. CA 125 sampling and a transvaginal pelvic ultrasound by a radiologist were systematically conducted every 6 months. Seventy-two patients were eventually evaluable. Two incident cases of peritoneal carcinomatosis (FIGO IIIB, malignant epithelial serous high-grade tumors) were discovered in two asymptomatic women with a deleterous BRCA1 mutation (2.7%). We did not observe any other primary cancer cases but an ovarian metastasis of a breast cancer. Forty women went off the study: 32 had a prophylactic bilateral salpingo-oophorectomy. Consistent with the literature, biannual screening tests combining CA125 and pelvis ultrasound is ineffective for early detection of a pelvic tumor of tubal or ovarian origin. Testing for BRCA1 or BRCA2 deleterious mutations is then crucial for suspected family syndromes of breast and ovarian cancer. For women carrying a deleterous mutation on BRCA1/2 a salpingo-oophorectomy is the only way, only the time of this surgery is debatable.

Colorectal cancers (CRC) express the epidermal growth factor receptor (EGF-R), a type I transmembrane receptor with tyrosine kinase activity. EGF-R signaling inhibition is a promising target for cancer therapy. ZD1839 (Iressa, AstraZeneca) and OSI-774 (Tarceva, Roche) are small molecular weight molecules with selective and reversible tyrosine kinase inhibition properties directed to EGF-R. Orally administered, these molecules induce sustained tumor stabilizations in previously treated metastatic CRC patients. The most frequent treatment-related toxicities are fatigue, diarrhea and acne-like follicular rash. The addition in the clinic of 5-FU, lOHP or CPT-11 to ZD1839 or OSI-774 does not seem to increase the own toxicity of each cytotoxic agents. Cetuximab (Erbitux, Merck) is an intravenously administered humanized monoclonal antibody which bind with high affinity with the extracellular domain of the EGF-R. The most frequent treatment-related toxicities are diarrhea, fatigue, nausea and cutaneous toxicity (allergic or acne-like follicular rashes, folliculitis). Most, if not all of these adverse events are mild. Partial responses were observed with cetuximab either alone (RR: 10%) or in combination with CPT-11 (RR: 22%) in patients with CPT-11 refractory advanced CRC which expressed EGF-R. The combination of cetuximab to folinic acid, 5-FU and CPT-11 seems tolerable at the cost of a slight increase of severe diarrhea and neutropenia. Finally, the promising activity of these EGF-R inhibitors has to be confirmed throughout randomized studies.

The proteome, first formalized in 1995, designs all the proteins expressed by the genome of a cell, tissu, or organ at a defined time. Proteomic analysis leads to a description of the regulation of gene expression by the study of proteins and of their post-translational modifications. Proteomic analysis is based on three technologies: 1) Two-dimensional electrophoresis allowing the separation of thousands of proteins from a single mixture; 2) mass spectrometry allowing the characterization of picoquantities of polypeptides and providing data on post-translational modifications; 3) Bioinformatic which is required for the quantification of protein level and for the constitution of databases of protein expression profiles. Complementing the methods of the genomics, the use of proteomic analysis is widely spreading in the fields of fundamental biology, biomedicine and pharmacology for the identification of new biological markers and therapeutic targets.

The BRCA1 gene modification is responsible for an autosomal dominant syndrome of inherited early onset breast and/or ovarian cancer. This gene is estimated to account for almost half of inherited breast cancers and three quarters of inherited breast/ovarian cancers. This suggests that about 1 out of 500 women may carry BRCA1 mutation. The BRCA1 gene was isolated by positional cloning in 1994. More than 100 different mutations have been found in the germline of affected individuals. We looked by systematic sequencing at BRCA1 germline mutations in 36 patients treated at the Centre Oscar-Lambret for breast and/or ovarian cancer and that belonged to high risk families. We have found 24 mutations: 9 true mutations inducing modifications of the BRCA1 protein (BRCA1+), 5 mutations with unknown consequences on the BRCA1 protein and 10 mutations corresponding to polymorphisms that had been previously described. All the BRCA1+ cases had a HPG3 tumor. The median age of discovery and the receptor positivity percentage are lower in hereditary breast cancer than in the standard population of the breast cancers treated in our center. Consequently, BRCA1 mutations are associated to parameters thought to be of bad prognosis.

Search for mutations of BRCA1 in women at hereditary risk for cancer is now possible. We asked the female gynaecologists of our county (north of France) their opinion about the search of a mutation of BRCA1 if they had a familial risk of breast cancer. Our aim was to obtain the opinion of informed women about their willingness to do the test for themselves and about the consequences they should accept. One hundred and eighty-three women received a questionnaire by post. The response rate was 56.3%. Twenty-four percent of the responders had a first degree relative with breast cancer. Most of the responders (87.4%; IC 95%: 81-93.8) would ask for the search of a mutation of BRCA1. The percentage of women who would accept the test is smaller for the women who have a first degree relative with breast cancer (72.0% vs 92.3%; P = 0.02). The reasons given to do the test were a better screening or prevention (69.7%) and the knowledge of a personal risk (49.4%). For breast cancer, 93.2% (95% CI: 88.4-98) would accept a screening protocol, 30.1% (CI: 21.3-38.9) would accept a prophylactic bilateral mastectomy. For ovarian cancer, 93.2% (CI: 88.4-98) would accept the screening, 52.4% (CI: 42.8-62) would accept a prophylactic ovariectomy. In conclusion, most of the informed women would ask for the test and the surgical options for reducing the risk of cancer are not absolutely rejected. Of course, only future studies will state precisely the choice of truly implicated women.

In this study, we have prepared antiidiotypic antibodies specific of human prolactin receptors (PRL-R) in order to localize these receptors in breast cancer. Antiidiotypic antibodies were prepared using anti-human prolactin (anti-hPRL) sera obtained from New Zealand rabbits. 25 breast cancer were PRL-R positive using radio receptor assay. For immunohistochemistry, the tissues fixed with Nakane and treated by NH4Cl and Saponin to improve staining specificity, were incubated with antiidiotypic antibodies (1/100e). Cytoplasm and membrane of human breast cancer cells were PRL-R positive in 20/25 breast cancer with antiidiotypic antibodies. There was no staining with preimmune serum and immunohistochemical staining elimination by prior absorption with anti-hPRL antibodies.

We investigated binding characteristics of bFGF in membranes prepared from 4 human breast cancer cell lines (MCF-7, T-47D, BT-20 and MDA-MB-231) and 38 primary breast cancer biopsies. Results of competitive binding experiments were analysed using the "Ligand" program to determine binding site concentrations and affinities. bFGF mitogenic activity was also measured by [3H]-thymidine incorporation into DNA of breast cancer cell lines. The presence of high-affinity binding sites was demonstrated in each cell type (Kd: 0.5 nM). The presence of these high-affinity binding sites was confirmed by saturation experiments. A second class of low-affinity binding sites was detected in the 2 hormono-independent cells (BT-20: Kd = 2.9 nM; MDA-MB-231: Kd = 2.7 nM). bFGF stimulated the proliferation of MCF-7, 7-47D, BT-20 and not of MDA-MB-231 cell lines. In breast cancer biopsies, binding sites were detectable in 36/38 cases; high-affinity binding sites (Kd < 1 nM) were present in 19/39 cases and low-affinity binding sites (Kd > 2 nM) were present in 29/36 cases (the 2 classes of binding sites were present in 12 biopsies). No relation between FGF binding sites and node involvement nature or grade of tumor was evidenced. Negative correlations (Spearman test) were found between total bFGF binding site concentrations and estradiol receptor concentrations (P = 0.05) or progesterone receptor concentrations (P = 0.009). The demonstrations of 1), bFGF specific binding sites in breast cancer membranes; and 2) bFGF growth stimulation of some breast cancer cell lines, indicate that this factor could be involved in the growth of most breast cancers, and could act (among other factors) directly on the growth of cancer cells.

The steroid receptor (ER, PgR) content of breast cancer specimens is now widely used as an indicator for hormone sensitivity which is a prerequisite for endocrine therapy. Is it conceivable to extend this steroid receptor determination to other types of tumors in order to select patients who would eventually benefit from an hormonal therapy? It seems that most normal or tumoral tissues contain detectable amount of steroid receptors. The values of each receptor comprise a continuum from very low to relatively high, suggesting that previous considerations of hormone dependence on the basis of presence or absence of hormone receptors may be oversimplified. The steroid receptor concentrations are however higher in target tissues, and could be of clinical interest in endometrium and prostatic cancers. The date are however insufficient to draw firm conclusions regarding their usefulness for treatment.

The authors have studied, on 100 cases, the prognostic value of ER and PgR receptor levels in cancers of the cervix. An elevated level of ER and PgR seems to favorably influence the immediate response to the treatment. On the contrary, it does not seem that this should be a factor in the long-term evolutive prognosis.

Prolactin (PRL) has an important peripheral role to play in female reproductive function. This hormone, in physiological concentrations, is necessary so that the follicle can mature properly. It also helps the maturation of oocytes and is essential for the physiological action of the corpus luteum so that progesterone production can be stimulated. It inhibits its catabolism and is responsible for keeping up the numbers of LH and oestradiol receptors. It works through the intermediary of prolactin receptors which are localised on the granulosa cells. The mechanism is through immuno-cytochemistry and it can be distinguished biochemically (Ka = 0.5 10(10) M). Probably, when levels are high it upsets follicular maturation by inhibiting the biosynthesis of the oestrogens and also by stimulating the secretion in the granulosa cells of a substance that inhibits oocyte maturation: and prolactin exercises a luteolytic action by stimulating the catabolism of the hormone and lessening the numbers of LH receptors. On the other and, there are high concentrations of PRL in amniotic fluid. As endometrial cells undergo the phenomenon of decidualization they produce PRL which is identical biochemically with the pituitary hormone. Certain steroids seem to regulate this synthesis of PRL. This hormone must play a role in fetal osmoregulation through the intermediary of prolactin receptors.

EGF membrane receptors (R-EGF) were assayed in 65 human breast cancer biopsies. The Scatchard analysis of the binding of 125I-EGF indicated a single class of sites with an apparent dissociation constant of 1 nM. Fourty-eight percent of the tumors are R-EGF positive (specific binding greater than 1% of total radioactivity), the level of receptors reaching 14%. We have not observed any relationship between R-EGF and hormone receptors (estradiol, progesterone and prolactin) or clinical and histological characteristics of the tumor. Taking into account the role of EGF in growth of some human breast cancer epithelial cells culture and the possible production of R-EGF by oncogenes, these results suggest that R-EGF could be an uindependent biochemical marker of human breast cancer.

Prolactin plays a peripheral role in male reproductive function just as it does in female function. Prolactin, through the medium of immunocytochemistry acts on testicular steroidogenesis through prolactin receptors which are sited on the Leydig cells. It alters the number of LH receptors and therefore the sensitivity of the testis to central stimulation and equally interferes with androgen synthesis. In the areas of physiological concentration the principal effect seems to be to stimulate secretion of testosterone by keeping up the number of LH receptors; but when there is acute hyperprolactinaemia, testosterone secretion is lowered or perhaps not changed at all because of a "post-receptor effect", in spite of there being larger numbers of LH receptors. The direct effect of prolactin on spermatogenesis has not yet been worked out and the results that have been obtained are controversial. Prolactin exerts a direct stimulating effect on the growth of prostatic cells working synergistically with testosterone through specific prolactin receptors. It could play a role in certain cancers of the prostate.