Overexpression of WT1 in all molecular subtypes of breast cancer and its impact on survival: exploring oncogenic and tumor suppressor roles of distinct WT1 isoforms.

BACKGROUND: Breast cancer is a highly heterogeneous solid tumor, posing challenges in developing targeted therapies effective for all mammary carcinoma subtypes. WT1 emerges as a promising target for breast cancer therapy due to its potential oncogenic role in various cancer types. Previous works have yielded inconsistent results. Therefore, further studies are needed to clarify the behavior of this complex gene in breast cancer. METHODS AND RESULTS: In this study, we examined WT1 expression in both Formalin Fixed Paraffin Embedded breast tumors (n = 41) and healthy adjacent tissues (n = 41) samples from newly diagnosed cases of ductal invasive breast cancer. The fold change in gene expression between the tumor and healthy tissue was determined by calculating 2-∆∆Ct. Disease-free survival analysis was computed using the Kaplan-Meier method. To identify the expression levels of different WT1 isoforms, we explored the ISOexpresso database. Relative quantification of the WT1 gene revealed an overexpression of WT1 in most cases. The percentage of patients surviving free of disease at 8 years of follow-up was lower in the group overexpressing WT1 compared to the group with down-regulated WT1. CONCLUSIONS: Interestingly, this overexpression was observed in all molecular subtypes of invasive breast cancer, underscoring the significance of WT1 as a potential target in all these subtypes. The observed WT1 down-expression in a few cases of invasive breast cancer, associated with better survival outcomes, may correspond to the down-regulation of a particular WT1-KTS (-) isoform: the WT1 A isoform (EX5-/KTS-). The co-expression of this WT1 oncogenic isoform with a regulated WT1- tumor suppressor isoform, such as the major WT1 F isoform (EX5-/KTS +), could also explain such survival outcomes. Due to its capacity to adopt dual roles, it becomes imperative to conduct individual molecular expression profiling of the WT1 gene. Such an approach holds great promise in the development of personalized treatment strategies for breast cancer.

Soluble MICA and anti-MICA Antibodies as Biomarkers of Nasopharyngeal Carcinoma Disease.

The MHC class I chain-related molecule A (MICA) is a ligand for the activating natural killer (NK) cell receptor NKG2D. A part from its genetic diversity, MICA is characterized by the presence of membrane-bound and soluble isoform (sMICA) and by the propensity to elicit antibody-mediated allogeneicity (MICA Abs). Altogether such properties are important in the cancer setting. Here, we investigated whether MICA polymorphism, serum level of sMICA and MICA antibodies (Abs) may influence nasopharyngeal carcinoma (NPC) risk. 274 NPC naïve of treatment patients and 275 healthy individuals, all originating from Tunisia were included and genotyped. Among them, 160 sera from patients and 51 from controls were analyzed for the sMICA level by ELISA and were tested for the presence of MICA Abs by Luminex assay. The statistical analysis showed that: (1) we extend and confer our previous finding concerning Val/Val association with risk of NPC (p = .02, OR = 1.56; 95%CI [1.12-2.11]). (2) The higher level of sMICA characterized patients advanced stage of the disease. (3) The 18 (78%) of patients having MICA Abs exhibit all a non-advanced stage of the tumor extension at presentation. MICA129 Met /Val, sMICA and MICA Abs could be potential biomarkers of prediction, the diverse staging of NPC and hence prognostic and treatment.

The Impact of HLA-G 3'UTR Polymorphisms in Breast Cancer in a Tunisian Population.

Human leukocyte antigens G and E (HLA-G and HLA-E) are nonclassical major histocompatibility complex (MHC) class I molecules. These molecules play an important role in immune surveillance by inhibiting natural killer and cytotoxic T cells responsible for immune escape. The expression of HLA-G and HLA-E has been associated with several diseases including tumor. The main objective of the study is to evaluate the impact of three HLA-G 3'UTR potential polymorphisms: +3187 A > G (rs9380142), +3142 G > C (rs1063320), +2960 14-base pair (bp) Insertion/Deletion (Ins/Del) (rs66554220), and the HLA-E*01:01/01:03 A > G (rs1264457) polymorphism in Tunisian breast cancer population. A total of 355 patients and 381 controls were genotyping for HLA-G and HLA-E polymorphisms using a Taq Man assay. +3142 C allele and +3142 C/C genotype were significantly associated with increased risk of breast cancer (p = 0.00002; OR = 1.58; 95% CI = 27-1.97) (49% versus 35%; p = 0.0001; OR = 1.79; 95% CI = 1.32-2.44). In addition, Del allele and the homozygous state for Del/Del genotype confer a risk for breast cancer (52% versus 45%, p = 0.006; OR = 1.33, 95% CI = 1.08-1.64) (28% versus 22%, p = 0.039; OR = 1.43, 95% CI = 0.90-2.25). However, no statistical significant differences were reported for HLA-G + 3187 A > G and HLA-E variations and breast cancer in a Tunisian population. The found results indicate that HLA-G may play an important role in the breast cancer.

Prognostic value of preoperative carcinoembryonic antigen level in colorectal cancer in Tunisia.

AIM: To evaluate the prognostic value of preoperative serum carcino-embryonic antigen (CEA) level in patients with colorectal cancer. METHODS: This retrospective study included 125 colorectal cancer patients aged from 14 to 87 years, surgically treated between January 2001 and December 2006. Preoperative serum CEA was measured by chemiluminescence assay. RESULTS: within the patients, 57 were males and 68 females. They have tumours classified Dukes A in 2 patients, B in 24 patients, C in 53 patients and Dukes D in 46 patients. Median follow-up period was 24 months (range, 4 - 72 months). The relapse-free survival was significantly higher in patients with CEA < 5 ng/ml compared to CEA ³ 5 ng/ml, (p < 0.0001). We observed significant differences in relapse-free survival between patients with CEA < 5 ng/ml and those with CEA ³ 5 ng/ml among patients classified as Dukes stage B (p=0.007) and C (p < 0.0001). However, there was no significant difference in relapse-free survival among those classified as Dukes stage D. Cox multivariate analysis demonstrated that preoperative serum CEA level was a significant independent prognostic factor for relapse-free survival (hazard ratio: 6.49, 95% CI, 3.09 to 13.62, p < 0.0001). CONCLUSION: Preoperative serum CEA is a reliable predictor factor for recurrence in patients with CRC. CEA might be used in staging system and will be useful for therapeutic orientation in patients undergoing curative resection of CRC.

C677t polymorphism of MTHFR and G80A polymorphism of RFC genes and their relation with homocysteine levels in obese Tunisian children.

AIMS: To investigate the frequencies of C677T polymorphism in MTHFR gene and G80A polymorphism in RFC gene in obese and no obese Tunisian children and to assess their relation with homocysteine (tHcy), folate and vitamin B12 levels. METHODS: We have studied 31 obese compared to 22 no obese children. tHcy was assessed by fluorescence-immunoassay ; folate and vitamin B12 by radioimmunoassay. C677T and G80A mutations were detected using pyrosequencing. RESULTS: There were no differences in tHcy levels between obese and no obese, (10,34 ± 4,86µmoll/l vs11,00 ± 4,26µmoll/l). We found no difference for the allelic frequencies of the C677T polymorphism (29.03 % vs 30.95 %) and of the G80A polymorphism (64.52 % vs 59.52 %). Mean levels of tHcy, folic acid and vitamin B12 were not significantly different according to MTHFR and RFC genotypes. CONCLUSION: We demonstrated no difference in tHcy, folates, vitamin B12 levels and allelic frequencies of C677T and G80A polymorphisms in MTHFR and RFC genes between obese and no obese Tunisian children. These two polymorphisms don't seem to have any impact on homocysteine, folate and vitamin B12 status in the two populations.

Deletion polymorphism of glutathione S-transferases M1 and T1 in the Tunisian population.

BACKGROUND: Glutathione S-transferases (GST) play a vital role in cellular defense against environmentally toxic compounds. These enzymes present a genetic deletion polymorphism, which varies with ethnicity. AIM: To evaluate the frequencies of homozygous deletion of GSTM1 and GSTT1 genes in Tunisian population. METHODS: On the basis of multiplex PCR protocol, the frequency of the deleted genotypes of GSTM1 and GSTT1 genes was evaluated on 145 healthy Tunisian subjects. RESULTS: We found that 34.6% of the individuals had the GSTM1 null genotype, 16.6% had the GSTT1 null genotype and 4.82% had a double deletion of both GSTM1 and GSTT1. CONCLUSION: The distribution of GSTM1 null in Tunisians is rather in the range of black populations and is lower than that reported in Asians, Arabs and Caucasians. However the frequency of GSTT1 null is in the range of several populations studied except Asians. The double deletion frequency seems lower than that described in different populations.

Use of common seric tumor markers in patients with solid cancers.

Seric tumour markers (STM) are molecules that theoretically indicate the presence of malignancy, and used for monitoring response to therapy and early detection of relapse. This article describes the use and limitations of common SMT in patient with solid tumors. Excepting prostate specific antigen (PSA) and thyroglobulin, STM are not poorly sensitive or specific for screening and diagnosis of cancers. Alpha foetoprotein (AFP) is however used to screen hepatocellular carcinoma in high risk patients with suspect masses. Beta subunit of human chorionic gonadotrophin (beta-HCG) is frequently used for the diagnosis and management of gestational trophoblastic disease, while combined AFP and beta-HCG dosage is a good adjunct in the evaluation and treatment of non seminomatous germ cell tumors. PSA is used for screening and follow up of prostate cancer. Ca 125 is useful for evaluating pelvic masses in postmenopausal women and monitoring response to therapy in women with ovarian cancer, while Ca 15 3 is used to follow response to therapy in patients with breast cancer.

[Erythropoietin in cancer patients with anemia]. / Erythropoietine chez des patients cancereux anemiques.

OBJECTIVE: The aim of this study is to evaluate the endogenous erythropoietin production in cancer patients with anemia. METHODS: Our prospective study interested 99 cancer patients with anemia from 17 to 80 years old, during the period going from March 2002 to December 2004, and 31 healthy individuals with anemia caused by iron deficiency. A blood sample was collected from each patient, as well as healthy individuals to measure serum erythropoietin, C reactive protein and ferritin. RESULTS: The increase of serum erythropoietin was significantly lower in patients than in healthy individuals (P < 0.05). 25.2% of our cancer patients have inflammatory anemia and 74.7% presented microcytic anemia associated with increase of serum ferririn and CRP. These values were significantly higher than in healthy individuals (p < 0.05). CONCLUSION: Anemia in cancer patients results from activation of inflammatory system, which inhibit erythropoietin production. Apart from etiologic treatments, anemia can be treated with recombinant human erythropoietin.

[Erythropoietin levels in perioperative period in cancer patients]. / Variation de l'érythropoïétine sérique chez des malades opérés en chirurgie carcinologique.

Anemia is frequent in cancer patients, is the result of decreased erythropoietin production. In fact in cancer, alteration of immune system alters iron metabolism and inhibits erythropoietin production. In this study we proposed to determine the profile of erythropoietin secretion in anaemic cancer patients in the pre and postoperative period. Our prospective study from January to March 2005 included 41 anemic cancer patients from 30 to 79 years old and 31 healthy individuals with iron deficiency anemia. A measure of erythropoietin, CRP, ferritin, iron levels and hemoglobin were released in healthy individuals and in cancer patients in preoperative period (J0) and postoperative period (J3, J8, J21). In preoperative period, the increase of serum erythropoietin was significantly lower in patients than in healthy individuals. In postoperative period, the levels of erythropoietin at J3 and hemoglobin's at J8 and J21 were significantly higher than in preoperative period (J0) (p < 0.05). In conclusion, despite the presence of inflammatory syndrome caused by surgery, cancer patients with anaemia increase their erythropoietin production in immediate postoperative period.

[Seric soluble interleukin-2 receptor alpha in nasopharyngeal carcinoma in Tunisia. Prospective study about 45 cases]. / Taux serique du recepteur alpha soluble de l'interleukine 2 dans le carcinome du nasopharynx en Tunisie. Etude prospective sur 45 cas.

BACKGROUND: Soluble interleukin-2 receptor alpha (slL-2Ralpha) is a well-known indicator of T-cell activation noted to be increasing in nasopharyngeal cancer. The aims of this study were to evaluate the importance of the use of this marker in nasopharyngeal carcinoma. METHODS: Our prospective study interested 45 patients (35M/10F) with a mean age of 49 years (15 to 78), presenting a nasopharyngeal carcinoma histologically confirmed and 61 healthy controls. A blood sample was collected from each patient before any treatment, as well as controls to measure sIL-2Ralpha by immunoenzymatic assay. According to the disease status after a period of follow-up ranging from three to 22 months (median 12 months), patients were divided into two groups: The remission group (n=28) represented those with favourable evolution and a second group of 15 patients with unfavourable evolution (2 death, 4 cases of persistent primary disease and 9 patients with distance metastasis). 2 patients were lost to follow-up. RESULTS: serum sIL-2Ralpha levels were significantly higher in patients vs healthy controls (p < 0.0001). The serum levels correlated with the stage T of NPC (p = 0.01). Patients having a favourable evolution have lower sIL-2Ralpha levels before treatment vs those with unfavourable evolution without statistical difference. CONCLUSION: Measurement of serum sIL-2Ralpha provides a good estimation of the nasopharyngeal tumor burden. The usefulness of this marker as a parameter to predict prognosis in NPC should be examined further.

[Genetic polymorphism of gluthation-S transferases and N-acetyl transferases 2 and nasopharyngeal carcinoma: the Tunisia experience]. / Polymorphisme génétique des enzymes gluthation-S transférases et N-acétyl-transférases 2 dans le cancer du nasopharynx en Tunisie.

Interindividual differences observed in the metabolism of xenobiotics have been attributed to the genetic polymorphism of genes, which code for enzymes involved in detoxification. This genetic variability seems to be associated with the individual's susceptibility to certain cancers, including nasopharyngeal carcinoma. In this study, we have investigated the genotypic frequencies of DNA polymorphisms of two detoxification's genes: the gluthatione-S-transferase (GST) and the N-acetyl transferase 2 (NAT2). The study has included 45 patients with nasopharyngeal carcinoma compared to 100 healthy Tunisian controls. The presence of the GSTM1 null and GSTT1 null polymorphism was screened by using a multiplex PCR procedure. A PCR-RFLP method was used to detect polymorphism for the most common alleles of the NAT2 gene. Allelic frequencies between the two groups were compared using a chi2 test and odds ratio with 95% confidence intervals were calculated. The results indicate that the genotypic frequency of GSTM10/0 between controls and patients was significantly different. This genotype confers an increased risk of nasopharyngeal carcinoma (Odds Ratio = 2.12, [0.64-4.7]). However, genotypic frequencies of NAT2*6/NAT2*6 were significantly higher in the group of nasopharyngeal carcinoma patients. The calculated Odds Ratio showed an association between this genotype and nasopharyngeal carcinoma. In conclusion, the increase of nasopharyngeal carcinoma risk in Tunisia seems to be associated with GSTM10/0 and NAT2*6/6 genotype.

[Nasopharyngeal carcinoma in Tunisian children: retrospective epidemiological, clinical and biological study about 48 cases]. / Le carcinome du nasopharynx chez l'enfant tunisien: étude rétrospective épidémiologique, clinique et biologique, à propos de 48 cas.

OBJECTIVE: Report epidemiological, clinical and biological aspects of nasopharyngeal carcinoma in Tunisian children. PATIENTS AND METHODS: Our retrospective study from 1994 to 2001 included all children treated for nasopharyngeal carcinoma in the Salah Azaïz Cancer Institute of Tunis. Initial investigation consisted of ENT and general examination, nasopharyngeal CT-scan, abdominal echography, chest X-ray and bone scintigraphy. Biological markers included blood-count, erythrocytes sedimentation and serum lactic dehydrogenase. All children received neoadjuvant chemotherapy (adriamycin, cisplatin) and irradiation therapy. RESULTS: There were 48 children with a median age of 13,7 years and a sex ratio of 1,4 (28/20). Lesions are staged T2, T3 and T4 in 2,1 %, 18,7% and 79,2% of cases. All patients have cervical palpable nodes at diagnosis classified as N1 (8,3%), N2 (33.3%) and N3 (58.3%). A significant correlation was found between serum lactic dehydrogenase and the N stage (p = 0.02). After follow up, recurrence of disease was noted for three children, persistent disease for two children and metastatic disease in five cases. The overall and relapse free survival at 5 years were 79.1% and 68.9% respectively. Patients aged 13 or lower had poorer 5 overall survival rate (72.3%) than older age group (84.2%).

[Contribution of serum Cyfra 21-1 in nasopharyngeal carcinoma in Tunisia]. / Apport du dosage sérique du Cyfra 21-1 dans le cancer du cavum en Tunisie.

Cyfra 21-1 is a recognised marker for epidermoid lung and head and neck carcinomas oriented to the cytokeratin 19 that is expressed particularly in malignant epithelial cells. The aims of this study were to evaluate the importance of the use of this marker in nasopharyngeal carcinoma (NPC). Our prospective study interested 41 patients (33M/8F) with a mean age of 44 years (13 to 70) with 8 of them aged less than 30 years, presenting a nasopharyngeal carcinoma histologically confirmed from September 1999 to March 2000 and 45 healthy controls without evidence neoplasm. Undifferentiated forms represent 90.2% of cases and lesions are staged T2, T3 and T4 in 2.4%, 36.6% and 61% of cases, while N1, N2 and N3 represent 9.8%, 26.8% and 41.5% of cases. A blood sample was collected from each patient and control before any treatment, as well as controls to measure Cyfra 21-1 by immunoenzymatic assay, 2 groups of patients were selected after a period varying from 4 to 37 months with a median of 29 months: 27 patients with favourable evolution (without evidence of disease after initial treatment), 12 patients with non favourable evolution (1 death, 2 cases of loco-regional relapse and 9 patients with metastatic disease). 2 patients were lost to follow-up. The results showed that the mean serum Cyfra 21-1 values were significantly higher in patients with NPC than those in controls (p = 0.001). A significant correlation was found between the serum Cyfra 21-1 level before treatment and the clinical outcome of patients (p = 0.0009). Patients having a favourable evolution have the lowest level. Seric level of Cyfra 21-1 at diagnosis of NPC may play a predictive role to evaluate the risk of metastatic disease and prognosis.