Maha Sallam

Detection of Epstein-Barr virus in breast carcinoma in Egyptian women.

BACKGROUND The association of oncogenic EBV with breast carcinoma (BC) is still in controversy. AIM OF WORK Assess the association of EBV with BC in Egyptian women and find possible relationship between prognostic factors of BC and EBV detection. SUBJECTS AND METHODS Paraffin-embedded sections from 40 female patients with primary invasive BC; ductal (n=32) and lobular (n=8) and breast tissues from patients with fibrocystic disease (n=20) as control were screened for presence of EBV by EBV nuclear antigen-1 (EBNA-1) immunostaining and by PCR for EBV-DNA. RESULTS 10/40 (25%) of the BC specimens stained positively for EBNA-1; EBNA-1 expression was restricted to a fraction 5%-60% of tumor epithelial cells. EBV-DNA was detected in 8/10 of BC specimens positive for EBNA-1. Control specimens were negative by both techniques. 7/8 (87.5%) of EBV-DNA positive tumors were associated with >3 lymph nodes involvement. CONCLUSION EBV is associated with some invasive BC in Egyptian females and may play a role in their etiology.

Increased bcl-2 expression is associated with primary resistance to chemotherapy in human epithelial ovarian cancer

OBJECTIVE bcl-2, an anti-apoptotic factor, has a role in the pathogenesis of ovarian cancer as well as in resistance to chemotherapy. DESIGN AND METHODS 20 benign, and 26 malignant epithelial ovarian tissues were analyzed for bcl-2 protein and mutant p53 by enzyme-immunoassay (EIA). Flowcytometric analysis was also performed. Patients of malignant group were followed up to monitor overall survival and primary resistance to chemotherapy. RESULTS bcl-2 was significantly higher in malignant group than benign group (p < 0.001). A cutoff value was determined for bcl-2 (63.8 kU/g protein). At this cutoff, sensitivity is 80.7%, and specificity is 85%. Using chi square analysis, a significant correlation was found between bcl-2 and FIGO stage (p = 0.01), overall survival (p = 0.01), as well as primary resistance to chemotherapy (p = 0.03). By correlation coefficient analysis the relation between bcl-2 and synthetic phase fraction was highly significant (p = 0.002). Bcl-2, p53, and FIGO stage were significantly correlated to poor survival (p = 0.01) in univariate analysis. However, in multivariate analysis, only FIGO stage, and p53 were independent risk factors. CONCLUSION EIA could be a useful tool for investigating the prognostic value of bcl-2, and its possible prediction of platinum resistance in epithelial ovarian cancer. This might help in selecting patients for future anti-bcl-2 therapy.

Preoperative serum inhibin levels in patients with ovarian tumors

Aim:  To assess the value of preoperative determination of serum inhibin levels in the prediction of malignancy in women with ovarian tumors. The prediction of malignancy not only helps patient counseling regarding prognosis and extent of surgery but also allows for proper specialist referral.

Laboratory validation of formal concept analysis of the methylation status of microarray-detected genes in primary breast cancer

Breast cancer is the leading cause of cancer-related mortality. DNA methylations play important roles in cancer development and progression. Formal concept analysis was previously utilized for data mining hypermethylated and hypomethylated genes in breast cancer molecular subtypes in illumina methylation–based microarray database, to laboratory validate their outputs; HS3ST2 (heparan sulfate d-glucosaminyl 3-O-sulfonyl transferase-2) and MUC1 (mucin-1) were retrieved. Both play important roles in progression and invasion of breast cancer. The methylation status of both genes was laboratory validated using methylation-based polymerase chain reaction in breast cancer subtypes luminal A (early stages) and luminal B (late stages) in comparison with benign conditions and normal breast to conclude their roles in tumor invasion and to validate the newly developed algorithm (formal concept analysis). Significant cancer-specific hypermethylation of HS3ST2 was detected in luminal B (chi square = 30.6, p = 0.000), while significant cancer-specific hypomethylation of MUC1 was detected in luminal B (chi square = 30.5, p = 0.001) breast cancer. The median levels of the percentage of methylated allele of both genes were significantly discriminative between luminal A and luminal B subtypes and benign and healthy control groups. Detection of MUC1 and HS3ST2 promoter methylation status appears to be useful molecular markers for assessing the progressive state of the disease and could be helpful in discriminating breast cancer molecular subtypes. These results validate the methylation-based microarray analysis, thus trust their output in the future.

KLK10 exon 3 unmethylated PCR product concentration: a new potential early diagnostic marker in ovarian cancer? - A pilot study

BackgroundKLK10 exon 3 hypermethylation correlated to tumor-specific lack of KLK10 expression in cancer cell lines and primary tumors. In the present study we investigate the possible role of KLK10 exon 3 methylation in ovarian tumor diagnosis and prognosis.ResultsQualitative methylation-specific PCR (MSP) results did not show statistically significant differences in patient group samples (normal and tumor) where all samples were positive only for the unmethylated-specific PCR except for two malignant samples that were either doubly positive (serous carcinoma) or doubly negative (Sertoli-Leydig cell tumor) for the two MSP tests. However, KLK10 exon 3 unmethylated PCR product concentration (ng/μl) showed statistically significant differences in benign and malignant patient group samples; mean ± SD (n): tumor: 0.077 ± 0.035 (14) and 0.047 ± 0.021 (15), respectively, p-value = 0.011; and normal: 0.094 ± 0.039 (7) and 0.046 ± 0.027 (6), respectively, p-value = 0.031. Moreover, ROC curve analysis of KLK10 exon 3 unmethylated PCR product concentration in overall patient group samples showed good diagnostic ability (AUC = 0.778; p-value = 0.002). Patient survival (living and died) showed statistically significant difference according to preoperative serum CA125 concentration (U/ml); median (n): 101.25 (10) and 1252 (5), respectively, p-value = 0.037, but not KLK10 exon 3 unmethylated PCR product concentration (ng/μl) in overall malignant patient samples; mean ± SD (n): 0.042 ± 0.015 (14) and 0.055 ± 0.032 (7), p-value = 0.228.ConclusionTo the best of our knowledge, this is the first report on KLK10 exon 3 unmethylated PCR product concentration as potential early epigenetic diagnostic marker in primary ovarian tumors. Taken into account the limitations in our study (small sample size and semi-quantitative PCR product analysis) further studies are strongly recommended.

Mitochondrial pathway of apoptosis and related proteins in placenta of pregnant Egyptian women with pre-eclampsia

Apoptosis within the placenta is increased in pregnancies complicated by pre-eclampsia (PE).This study aimed at evaluating the mitochondrial pathway of apoptosis in placenta of pregnant women with pre-eclampsia and correlate it with severity and pregnancy outcome . Apoptosis was assessed by measuring DNA fragmentation%,Bcl-2, p53 and caspase-9 in placental tissues from 25 pregnancies complicated by pre-eclampsia, and 25 placentas of normal pregnancy (NP). DNA fragmentation, p53 and caspase-9, were significantly increased while, Bcl-2 was significantly decreased in the placenta of pre-eclampsia group compared to control. No significant difference between mild and severe pre-eclampsia subgroups regarding these parameters could be found. DNA fragmentation was negatively correlated with Bcl-2 in PE group. Moreover, DNA fragmentation was negatively correlated with maternal age in both PE and control groups. A significant positive correlation between placental DNA fragmentation and gestational age in the NP group was observed. A significant negative correlation between caspase-9 activity, and fetal weight in PE group was found. It can be concluded that Hypoxia and ischemia induced by pre-eclampsia, up regulate p53 and reduce Bcl-2 expression with activation of caspase-9 and increasing DNA fragmentation in placental tissue. These results implicate the mitochondrial pathway in enhancing apoptosis in preeclamptic placenta and affecting fetal outcome but not the severity. Keywords : Apoptosis, p53, Bcl-2, Caspase-9, Mitochondria, Pre-eclampsia.