Evaluation and clinical correlation of bone marrow angiogensis and level of serum angiogenic factors in acute leukemia

In this study serum angiogenic factors vascular endothelial growth factor [VEGF], hepatocyte growth factor [HGF] and tumour necrosis factor a [TNF a]] and cellular angiogenic factors [VEOF and VEGF-R2] were studied in 50 newly diagnosed acute leukemia patients, they were 24 ALL and 26 AML patients. The correlations of the studied angiogenic factors to each other and to the patients' survival and disease outcome were studied. During the follow-up period of 6 months, 22 patients died and 28 patients remained alive from whom 11 patients were refractory and 17 patients achieved complete remission. On comparison between pretreatment concentration levels of measured serum angiogenic factors [VEOF, TNF-alpha and HOF] in ALL, AML and the control group, all the comparisons were statistically significant [p<0.0001, <0.0001 and 0.02 1 respectively]. All serum markers were higher in AML group than control group, but only VEOF showed statistically significant elevation [p<0.0001], while in ALL patients, all markers were significantly higher than control group [p=0.01]. When comparing ALL and AML cases according to cellular angiogenic factors detected by immunocytochemistry, cellular VEGF-R2 was slightly higher in ALL group, while cellular VEGF was slightly higher in AML group. The comparisons were statistically non-significant for both angiogenic factors. As regards response to therapy, in ALL, cases with high sVEGF showed a statistically significant lower rate of complete remission than cases with low sVEGF [p=0.041]. The same results were obtained for AML but the comparison did not reach a significant level [p=0.082]. Serum VEOF was the only reliable marker to predict relapse in ALL [p=0.009] and AML [p=0.049]. On comparing serum VEGF to the outcome in ALL, high sVEGF cases showed a statistically significant higher rate of death than low sVEGF cases [prO.05], while in AML, the same results were obtained but the comparison did not reach a significant level. As regards the survival time, cases with low sVEGF level showed higher mean survival and 6-month survival than cases with high sVEGF level p=0.03]. A significant negative correlation was detected between serum VEGF and serum TNF-a [correlation coefficient [r]=-0.642, p<0.0001]. Conclusion: Serum angiogenic factors [VEGF, TNF-alpha and HOF] are markedly increased in cases of acute leukemia compared to normal controls. Cases with high sVEGF showed higher rate of death than cases with low sVEGF, so its targeting may provide a potent novel therapeutic approach in acute leukemias. VEGF may also be useful as a new prognostic factor and a predictor of relapse in different types of acute leukemia. Further studies with larger number of patients and longer duration of follow-up are recommended to throw more light on the significance of other angiogenic factors in relation to acute leukemia

Study of the expression of survivin and mutant p53 in pediatric acute lymphoblastic leukemia

Childhood acute lymphoblastic leukemia is the most common malignancy in children with a yearly incidence of 31/1000000 children younger than 15 years old. The peak incidence of childhood ALL is between 3 and 6 years of age with male predominance. The relative frequency of pediatric ALL in the NCI-Cairo University is 35.5% for the years 2003-2004. In this study survivin and mutant p53 expressions were studied in 64 newly diagnosed pediatric ALL patients. Their associations to the different prognostic factors of ALL and their association to each other were studied. In this study, 21 out of 64 ALL cases [32.8%] showed positive expression of survivin [17 patients were moderately positive, 2 patients were strong positive and 2 patients were weak positive] and 24 out of 64 studied ALL cases [37.5%] demonstrated positive expression of p53 [20 patients were strong positive, 3 patients were moderately positive and 1 patient was weak positive]. On comparing survivin expression with the different prognostic factors of ALL, the results were statistically significant as regards the percentage of blasts in the peripheral blood [p-value = 0.0068], and the percentage of blasts in the bone marrow [p-value = 0.05]. As regards LDH, ALP, and uric acid serum concentrations, no statistically significant differences were found [p-value = 0.154, 0.52 and 0.41 respectively]. No significant association was found between survivin expression and hepatosplenomegaly. As regards p53 expression compared with the different prognostic factors no statistically significant results were found. According to immunophenotyping [IPT], survivin was positive in 5 out of 14 cases [35.7%] of proB-ALL, 0% [0/11] C-ALL, 29.2% [7/24] preB-ALL, 75% [6/8] mature B-ALL and 42.9%[3/7] T-ALL. The results were statistically significant [p-value = 0.046], while p53 was positive in 4 out of 14 cases [28.5%] of proB-ALL, 36.4% [4/11] C-ALL, 41.6% [10/24] preB-ALL, 50% [4/8] mature B-ALL and 28.6% [2/7] T-ALL. The results were statistically non significant. No significant correlation was found between survivin and p53 expression in the studied ALL cases [p-value = 0.872]. In this study, a total of 23 patients successfully completed induction phase. All of them achieved complete remission. Two patients developed isolated bone marrow relapse at a median period of 7.5 months. The disease free survival for the 23 patients was 89.6% at a median of 11 months. The DFS for P53 positive [12/23] and p53 negative [11/23] patients was 91.7% and 90.9% respectively [p= 0.90]. The DFS for survivin positive [11/23] and survivin negative [12/23] patients was 85.7% and 93.7% respectively [p=0.49]. In conclusion, we could not find any association between p53 and survivin expressions and the different prognostic factors of pediatric ALL patients, [the only statistically significant results were obtained when comparing the blast count% in both the peripheral blood, and the bone marrow between survivin positive and survivin negative cases]. As regards the comparison of survivin expression and phenotyping of the studied patients, it was not expressed in C-ALL cases which are known to have a good prognosis. Further we could not decide whether positive p53 or survivin in ALL patients had an impact on DFS. Further studies with larger number of patients and longer duration of follow up are recommended to throw more light on the significance of p53 and survivin in relation to ALL patients

study into the expression of the tumor suppressor gene p53 and the oncogene bcl-2 in human breast cancer

In this study, the protein expression of the mutant tumor suppressor gene p53, and oncogene bcl-2 was evaluated in 50 female breast cancer patients using two different techniques, ELISA and western blotting, the relation between these two techniques in relation to protein expression was detected, and the correlations between the expression of these two proteins and the different prognostic parameters of cancer breast were studied. The mean value of bcl-2 in breast cancer patients was statistically significantly higher than those of normal healthy controls using both ELISA and western blotting techniques. A significant positive correlation was found between bcl-2 and both the tumor size and the stage of the tumor which signifies its important role as a prognostic factor in breast cancer. Also over-expression of mutant p53 was detected using both ELISA and western blotting techniques when compared to the normal healthy controls. The only statistically significant correlation between mutant p53 expression and the different prognostic parameters of breast cancer was found to be between p53 the stage of the tumor. When taking predictive cut off levels of bcl-2 at 75U/ml, and mutant p53 at 2.16U/ml, bcl-2 showed the highest sensitivity, specificity anti diagnostic accuracy of [88%, 90%, and 90%] respectively, while mutant P53 showed its highest sensitivity, specificity and diagnostic accuracy [80%, 100%, and 85.7%] at the chosen cut offs. When done in either abnormal and both abnormal double combinations with the same cut offs, the sensitivity, specificity and diagnostic accuracy in the either abnormal combination have improved than either parameter singly, [92%, 100% and 94.29%] respectively, while in both abnormal combination the sensitivity, specificity, and diagnostic accuracy are [64%, 90% and 71.43] respectively. When studying the correlation between the 2 techniques [ELISA and western blotting] used in determination of bcl-2 and P53 expression in breast cancer patients all samples which showed over-expression for mutant P53 and bcl-2 using ELISA technique, also demonstrated over-expression using western blotting technique. The percent of positive expression the mutant P53 and the bcl-2 proteins were higher [76%] and [88%] respectively using western blotting technique when compared to ELISA technique [70%] and [80%] respectively. The results of both techniques were statistically significantly correlated for both P53 and bcl-2. Although western blotting technique has a higher sensitivity compared to ELISA, it has many disadvantages as cost, reproducibility, time consumption and the requirement for high skills

Study of angiogenic factors: Vascular endothelial growth factor [VEGF] and basic fibroblast growth factor [bFGF] in hepatocellular carcinoma

Hepatocellular carcinoma [HCC] is one of the most commonmalignancies in the world. Hepatocellular carcinoma is characterized by high vascularity, so tumor angiogenesis nowadays has been considered to be a strong prognostic factor in patients with HCC .The pre-therapeutic serum vascular endothelial growth factor[VEGF] and basic fibroblast growth factor [bFGF] levels in the HCC patients appear to reflect the disease's potential activity of vascular invasion and metastasis. The pre-therapeutic serum levels of the angiogenic factors VEGF and bFGF were detected in the sera of HCC patients to find new markers to be used for diagnosis of HCC, and were compared with the routinely used tumor markers used for diagnosis of HCC as AFP, CEA, and CA19.9. The relation between the serum levels of VEGF, and bFGF and the clinical characteristics of HCC was also elucidated. On comparing the studied tumor markers among the three studied groups, all the tumor markers were highest in the HCC group, followed the benign liver diseases, and lastly the normal control group [p-value= <0.001 each]. On comparing the studied tumor markers according to different prognostic factors, only AFP showed statistically significant result with the tumor size