Evaluation of genotoxicity of Euphorbia triaculeata Forssk. extract on mice bone marrow cells in vivo.

Chemotherapeutic agents for cancer are highly toxic to healthy tissues at therapeutic doses and hence alternative medicine avenues are widely researched. Most of the studies on alternative medicine have suggested that Euphorbia plant possesses considerable antitumor and antibacterial properties. The present study was designed to evaluate the in vivo genotoxic effects of Euphorbia triaculeata extract on mice bone marrow cells using chromosomal aberration test and micronucleus assay. This study also deals with the effect of E. triaculeata on the standard drug cyclophosphamide (CP) treatment in mice. Three different doses 250, 500 and 1000 mg/kg body weight were selected. In micronucleus assay, single oral dose administration of Euphorbia triaculeata extract at the three doses did not increase the number of micronucleated polychromatic erythrocytes. Similarly, a single oral administration of Euphorbia triaculeata extract showed no significant changes on mitotic indices or in induction of chromosomal aberrations in mice bone marrow cells. Pretreatment with E. triaculeata extract significantly reduced the clastogenicity of CP. Hence it can be concluded that, E. triaculeata extract showed no significant genotoxic effect on mice bone marrow cells. Under the conditions of this study, it has been demonstrated that the Euphorbia triaculeata extract is not genotoxic and not clastogenic at the concentrations used.

Genetic alterations in benign prostatic hyperplasia patients.

Background: Benign prostate hyperplasia (BPH) is a classical age-related disease of the prostate, present in 20% of men at the age of 40 years with progression to 70% by the age of 60 years. BPH is associated with various lower urinary tract symptoms, which affect their day-to-day life. Materials and methods: Our objective was to evaluate the association between HER-2/neu, c-myc, p53, and clinicopathological variables in 45 patients diagnosed with benign prostatic hyperplasia using fluorescence in situ hybridization (FISH). The patients underwent transurethral prostate resection to address their primary urological problem. All patients were evaluated by use of a comprehensive medical history and rectal digital examination. The preoperative evaluation also included serum prostate specific antigen (PSA) measurement and ultrasonographic measurement of prostate volume. Results: The mean (± standard deviation) age of the 45 patients was 69.65 ± 8.97 years. The mean PSA value of the patients was 9.25 ± 5.12 ng/mL. The mean prostate volume was 65.46 ± 11.43 mL. Amplification of HER-2/neu was seen in 4/45 (8.9%) cases and amplification of c-myc was seen in 5 of 45 (11.1%) cases; both genes were not associated with adverse clinicopathological variables. Deletion of p53 was seen in 20/45 (44.4%) cases. p53 gene was significantly associated with a severe AUASI (American Urological Association Symptom Index) score. Conclusion: In this study, we discussed important genetic markers in benign prostatic hyperplasia patients which may, in the future, be used as markers for diagnosis and prognosis, as well as targets for therapeutic intervention.

Evaluation of Cytotoxic and Genotoxic Effects of Euphorbia Triaculeata Forssk. Extract

Objective: To evaluate the cytotoxic and genotoxic activity of Euphorbia triaculeata Forssk. plant extract from Jazan region, Saudi Arabia, in an in vitro cancer model, which could be beneficial in anticancer therapy against human breast cancer cell line (MCF-7), prostate cell line (PC-3), human hepatocellular carcinoma cell line (HEPG2) and normal breast epithelial cell line (MCF-10A). The human foreskin fibroblast cell line, (Hs68), was also included in the cell panel. Doxorubicin and 5-Flurouracil, broad-spectrum anticancer drugs, were used as the positive control. Methods: Cytotoxicity of Euphorbia triaculeata plant extract was investigated by employing MTT assay and the genotoxicity was assessed by using comet assay. Results: Both toxicity tests exhibited significant toxicity results. In the comet assay, the Euphorbia triaculeata extract exhibited genotoxic effects against MCF-7 DNA and PC 3 but not on HEPG2 cell lines in a time-dependent manner by increasing the mean percentage of DNA damage. Euphorbia triaculeata extract showed significant toxicity against cancer cells. Comparison with positive control signifies that cytotoxicity exhibited by methanol extract might have moderate activity. Conclusion: The present work confirmed the cytotoxicity and genotoxicity of Euphorbia triaculeata plant. However, the observed toxicity of this plant extract needs to be confirmed by additional studies. Based on our results, further examination of the potential anticancer properties of Euphorbia triaculeata plant species and the identification of the active ingredients of these extracts is warranted.

Chromosomal aberrations in early-stage bilharzial bladder cancer.

Bilharzial bladder cancer is one of the most common types of malignancy in both men and women in several developing countries including Egypt. It has several unique clinical, epidemiological, and histological characteristics, suggesting that it is an entity distinct from bladder cancer seen in Western countries. Genetic alterations in bilharzial-related bladder cancer have been studied infrequently, especially in the advanced stages of disease, that is, T3 and T4 classifications. The objective of this study was to extend establishing the baseline cytogenetic profile of this type of malignancy to early T1 and T2 classifications. For this purpose, fluorescence in situ hybridization was applied to interphase nuclei of frozen-stored samples with biotinylated repetitive DNA probes specific for all chromosomes to detect numerical chromosome changes in 35 patients presenting with relatively early-stage pT1 and pT2 disease. Eleven cases had squamous cell carcinoma (SCC) and 24 had transitional cell carcinoma. Six of 24 transitional cell carcinomas had diploid chromosome counts with all the probes. Numerical chromosome aberrations were detected in 18 cases (75%). In 12 cases, a loss of chromosome 9 was observed. In three cases, an additional loss of chromosome 17 was detected. One case demonstrated a loss of chromosome 10, whereas another two cases showed a gain of chromosome 7, next to a loss of chromosome 9. Loss of chromosome Y was observed in nine of the 27 male cases studied (33.3%), in which only one case showed an abnormality whereas four cases were detected next to loss of chromosome 9, and one case showed gain of chromosome 7. Five cases showed loss of chromosome 19 whereas gain of chromosome 4 was detected in two cases. Two of 11 samples of SCC had normal diploid chromosome counts with all the probes used. In four of 11 cases (36.4%) underrepresentation of chromosome 9, compared with the other chromosomes, was detected. An additional loss of chromosome 17 and gain of chromosome 7, next to loss of chromosome 9, was detected in three cases. One case showed loss of chromosome 17 as the only numerical aberration. Loss of the Y chromosome was detected in three cases of which one case had gain of chromosome 7 and one case had loss of chromosome 19. No correlation was found between any of the clinicopathologic parameters examined in this study and the presence or absence of any numerical chromosomal aberrations except for the significant association between schistosomal history and loss of Y chromosome (P=0.007).

Detection of C-erb B2 gene amplification in bilharzial associated bladder cancer using fluorescence in situ hybridization.

BACKGROUND: Gene amplifications are common events in different tumor types and may confer diagnostic, prognostic, or therapeutic information for patient management. Fluorescence in situ hybridization (FISH) represents a standard methodologic approach for testing for this genetic alteration, as it is rapid, reproducible and extremely reliable in detecting presence of C-erb-B2 gene amplification for clinical utility. PATIENTS AND METHODS: In this study, FISH is used in a series of archival human bilharzial bladder cancer specimens to evaluate for the presence of cerbB-2 gene alterations in the most common malignant tumor in bilharzial endemic areas, e.g., Egypt and some other countries. The study included 40 cases, 30 males and 10 females. Their ages ranged between 30 years and 76 years (median: 51 years). Twenty-one cases had squamous cell carcinoma, 16 had transitional cell carcinoma, two had adenocarcinoma, and one case had undifferentiated carcinoma. RESULTS: Thirteen out of 40 tumor samples (32.5%) show evidence of true C-erb-B2 gene amplification. Of the remaining samples, 24 (60%) show no gene amplification and three (7.5%) fall into the borderline category with a ratio between one and two C-erb-B2 genes/cell relative to chromosome 17 centromeres. No evidence of chromosome 17 polysomy was found in any cases scored as single copy with the C-erb-B2 probe. CONCLUSION: No significant association was found between gene amplification and any of the tested clinicopathologic parameters or tumor recurrence except for tumor grade where higher tumor grades tended to be associated with more C-erb-B2 gene amplification (P = 0.01) thus reflecting more tumor aggressiveness. So, the amplification of C-erb-B2 in bilharzial associated bladder cancer is probably not independently related to clinical outcome of patients.

Chromosomal aberrations in Cis and Ta bilharzial bladder cancer: a theory of pathogenesis.

BACKGROUND: Bladder cancer manifests many different forms ranging from superficial to aggressive muscle invasive stages, which suggests that various genetic alterations are involved. Several attempts have been made to establish correlations between specific genetic alterations and various stages of the disease. At the National Cancer Institute (NCI), Cairo, bladder cancer constitutes 30.3% of all cancers. Bladder cancer observed among Egyptians has a clinico-pathological profile that differs from transitional cell carcinoma (TCC) seen in the western world. PATIENTS AND METHODS: We used fluorescence in situ hybridization to detect numerical chromosome changes in 25 patients presenting with carcinoma in situ and Ta lesions. Twenty-four cases had transitional cell carcinoma and one case had squamous cell carcinoma. RESULTS: Five out of 24 TCC cases had diploid chromosome count with all the probes. Numerical chromosome aberrations were detected in 19 cases (79%). In eight cases, a loss of chromosome 9 was observed. In one case, an additional loss of chromosome 17 was detected. One case demonstrated a loss of chromosome 17, whereas another three cases showed a gain of chromosome 7. Loss of chromosome Y was observed in nine of the 22 male cases studied (40.9%). The only case with SCC had normal diploid chromosome count with all the probes used. CONCLUSION: When the genetic basis of bilharzial related bladder cancer is fully understood, new diagnostic and therapeutic strategies will be developed, which in turn may promote better clinical management by pathologists and urologists. A theory of bilharzial related bladder cancer pathogenesis is suggested.

Cytogenetic profile of locally advanced and metastatic Schistosoma-related bladder cancer and response to chemotherapy.

Bladder cancer is a common malignancy in developing countries in which bladder infection with the parasite Schistosoma haematobium is prevalent. Several epidemiological, histopathological, and clinical characteristics of schistosoma-associated bladder cancer suggest that it is distinct from bladder cancer seen in other places in the world. The aim of this study was to extend establishing the cytogenetic profile of this type of malignancy in advanced and metastatic cases, and to demonstrate its relation to the end results of systemic therapy. Fluorescence in situ hybridization was applied to interphase nuclei to detect numerical chromosome changes in 41 patients with bladder cancer. Numerical chromosome aberrations were detected in 27 of 41 cases (66%). In 17 (41%) cases, a gain of chromosome 7 was observed, while losses in chromosomes 9 and 17 were detected in 20 (49%) and 18 (44%) cases, respectively. Loss of chromosome Y was detected in 7 of the 32 male patients included in this study (22%). There was a statistically significant association between stage of the disease and overall survival; Bajorin score and time to disease progression and overall survival; and between response to systemic therapy and time to disease progression and overall survival. The only chromosomal abnormality that had a significant relationship with overall survival was the gain of chromosome 4. When the genetic basis of schistosoma-associated bladder cancer is fully understood, new diagnostic and therapeutic strategies could be developed, which in turn may promote better clinical management and survival.

Detection of HER-2/neu, c-myc amplification and p53 inactivation by FISH in Egyptian patients with breast cancer.

Breast cancer is a leading cause of cancer-related deaths in women worldwide. The clinical course of this disease is highly variable and clinicians continuously search for prognostic parameters that can accurately predict prognosis, and indicate a suitable adjuvant therapy for each patient. Amplification of the two oncogenes HER-2/neu and c-myc and inactivation of the tumor suppressor gene p53 are frequently encountered in breast carcinomas. The purpose of this study was to use the fluorescence in situ hybridization (FISH) for the assessment of HER-2/neu and c-myc amplification and p53 inactivation and to relate these molecular markers with the commonly used clinical and pathological factors. The study was conducted on 34 tissue samples obtained from 33 females and 1 male with breast carcinomas and 17 samples obtained from 16 females and 1 male with benign breast lesions. Results revealed that the level of HER-2/neu, c-myc and p53 in the malignant group was significantly increased as compared to the benign group. On relating the level of the molecular markers to clinicopathological factors, p53 was significantly associated with increased patient's age. The sensitivity of the investigated markers significantly increased with larger tumor size. Concerning tumor grade, HER-2/neu and p53 showed a significant increase in low-grade tumors whereas c-myc showed a highly significant increase in high-grade tumors. With regard to disease staging, HER-2/neu and c-myc were the only markers that showed significant increase at late stages of disease. p53 and HER-2/neu were significantly associated with positive lymph nodal status. A significant correlation was obtained between the levels of the three biomarkers to each other. Conclusively, the combination of HER-2/neu, c-myc and p53 can stratify patients into different risk groups.