Reverse transcriptase-PCR detection of Egyptian breast cancer micrometastasis using multi marker panel

There is a growing body of evidence that cancer cells infiltrate peripheral blood in very early stages of the breast cancer. Only about one per 105-106 cancer cells will get to and settle in distant organs and a small percentage of those will develop metastasis. The aim of this study was to use Four marker assays for detection of breast cancer cells circulating in patients' blood. We analyzed the usefulness of four mRNA markers: Mammaglobin [hMAM], epidermal growth factor receptor [EGFR], matrix metal loproteinase-2 and -9 [MMP-2, MMP-9] using RT-PCR technique. Blood samples from 135 cases [75 primary breast cancer patients and 60 healthy volunteers] were tested for the previously mentioned markers. hMAM transcripts were found in 41 of 75 cases [55.6%] of malignant patients and in 1.66% of the healthy donors [p=0.0001]. EGFR transcripts were found in 42.6% of malignant patients and 3.3% of healthy volunteers [p=0.00001]. MMP-2 transcripts were detected in 38.6% of malignant cases and not detected in healthy subjects [p=0.0001]. In case of MMP-9, no significant difference was observed between all cases studied [p=1.00]. No significant correlation was existed among molecular markers studied and clinicopathological data except for MMP-2 with ER positive vases [p=0.012]. These data suggest that, hMAM, EGFR, and MMP-2 allowed the detection of circulating breast cancer cells and MMP-2 is more specific to tumor and is associated with ER positive breast cancer cases

Early detection of Egyptian colorectal cancer patients using stool-based DNA markers

About 50% of the patients with colorectal cancer who are diagnosed on the basis of clinical symptom could be cured by surgery. It may be assumed that diagnosis and surgery at an earlier asymptomatic stage would allow more patients to be cured. Therefore, screening appears to be the simplest way to decrease mortality due to colorectal cancer. The emergence of molecular stool testing promises accurate and user friendly alternative to conventional methods of colorectal cancer screening. The aim of this work is to evaluate the stool-based DNA test for three genetic markers on Egyptian patients. DNA was extracted from stool and/or tissue samples collected from 45 Egyptian patients [17 colorectal cancer, 28 non-cancerous lesions and normal] the lesions include ulcerative colitis, polyps, and schistosomiasis. Mutations were detected in k-ras oncogene at codon 12 using RFLP technique, p53 gene [exons 5, 6, and 7] using SSCP technique and finally microsatellite instability [MSI] for BAT-26 locus as a marker for mismatch repair gene. The overall molecular changes using the three markers showed that 25 patients out of 45 [55.56%] exhibited mutations in one or more of the markers used. Mutations were detected in 76.46% colorectal cancer patients. In non-cancerous cases, 42.86% exhibited mutations using the same gene panel. P53 mutation recorded the highest percentage of markers used [58.8%] in cancerous cases whereas it was 42.86% in non-cancerous cases. K-ras mutations recorded about 29.4% in cancerous cases and non in non-cancerous cases. Finally, MSI at BAT-26 was only 5.9% in cancerous cases and no MSI in non-cancerous group. In conclusion, p53 may represent a golden marker for Egyptian patients with colorectal cancer and pre-malignant diseases. Using multi-target genetic panel is successfully for early detection of colorectal cancer. Finally, stool-based DNA test is a real golden method which reflect the genetic changes in the tumors

Loss of heterozygosity at BRCA1, TP53, nm-23 and other loci on chromosome 17q in human breast carcinoma

In Egypt, breast cancer ranks number one among the female malignancies. Activation of oncogenes and inactivation of tumor suppressor genes are thought to play an important role in the development and progression of breast cancer. The present study is a trial to investigate the role of chromosome 17 in sporadic invasive ductal carcinoma of the breast through detection of LOH for 6 highly polymorphic microsatellite loci, two of which are located at BRCA1 gene [D17S855 and D17S856], one at TP53 gene, one at nm 23 gene and finally two at 17q12-12.3 [D17S183 and D17S250]. Tissue samples and their corresponding safety margin normal tissues were collected from 25 patients with invasive ductal carcinoma of the breast of grades 2 and 3. LOH was detected for the 6 highly polymorphic microsatellite markers mentioned previously using PCR assay. The percentage of overall LOH recorded was 68% of the cases examined. The highest LOH was recorded in D17S855 and D17S856 [43% and 32% respectively], both markers are located at BRCA1 gene, followed by 32% LOH in nm-23 gene. D17S183 and D17S250, which are localized telomeric and centromeric to BRCA1 gene, showed 24% and 28% LOH, respectively. The lowest percentage of LOH was observed in the TP53 gene [14%]. No significant correlation was found between each of the six markers used and lymph node status, grade, or menopause status. LOH at the nm-32 marker exhibited a significant association with lymph node involvement. It can be concluded from the present study that BRCA1 gene may be involved in carcinogenesis of some sporadic breast cancer cases. Deletion in nm-23 gene is associated with the advanced stage of the disease. Finally, another gene located at 17q 12- 12.3 region may be involved in some sporadic breast cancer cases

P53/WAF1 dependent pathway in squamous cell carcinoma of bladder associated with Bilharziasis

Our understanding of cellular growth control and cell cycle control has been studied extensively at the last decade. Several genes have been identified whose expression are affected by p53 gene. The waf1 is a target gene whose expression has been found to be regulated by p53. We studied p53/waf1 expression inschistosomal egyptian bladder cancer patients in an attempt for the possible use of them as genetic marker for assessment of the progression of such type of disease in egypt. For this purpose, 50 bladder tumors and their corresponding safty margins as controls were collected from nci, cairo niversity and screened for detection of p53 alterations using western blotting confirmed by pcr-sscp analysis, as well as the expression of waf1 gene using western blotting technique. Our results revealed that 20 cases [40%] showed mut-p53, 19 of them harbor abnormal pattern of sscp analysis. A significant correlation was reported between p53 and histopathological type of bladder cancer [scc and tcc, p=0.02]. We reported 21 cases [42%] showed waf1 downexpression. This study gives a dramatic example for the interaply between the oncosuppressor genes in cancer where, a very strong negative association was reported between the presence of p53 alteration [at both dna and protein levels] and waf1 expression [p=0.001]. In conclusion, this study provides an evidence that p53 may be involved in scc genetic. However, no role for p53 alterations in the progression or metastatic potential of bladder cancer. Also, p53/waf1 deficiency may not involve in bladder cancer development and/or progression