Evaluation of hepatocyte growth factor [HGF] as a prognostic indicator in acute myeloid leukemia

This study evaluated the serum levels of hepatocyte growth factor [HGF] and analyzed their prognostic significance in patients with acute myeloid leukemia. Enzyme linked immuno-sorbent assay [ELISA] was performed to quantify the HGF in stored samples obtained from 40 patients with acute myeloid leukemia [AML] at diagnosis and reanalysis was performed in those who obtained complete remission [CR]. Real time PCR was earned out on thirteen samples that were selected as having the lowest ELISA readings for HGF after complete remission. We aimed to study the potential role of HGF in prognosis and severity of AML. We investigated the other currently available methods for AML diagnosis including bone marrow morphology, cytochemistry, immunophenotyping and cytogenetics for all patients. Our data showed that the levels of HGF were significantly higher in patients with AML than in healthy individuals [4870.52 +/- 6310.48 Vs 410.2 +/- 185.89 with p<0.001] with a significant reduction of HGF levels after achieving complete remission [341.09 +/- 181.05 with p<0.001]. A significant correlation between HGF level and total leukocytic count [TLC] [p 0.001], bone marrow blasts [p 0.02] and time to complete remission [p 0.03] was found. We found no correlation between TLC and time to complete remission [p 0.32], but there was a poor correlation between bone marrow blast% and time to complete remission [p 0.17]. Only three samples out of the thirteen ones that were subjected to RT-PCR detection of HGF-mRNA showed mRNA down regulation, while the rest of cases did not express HGF mRNA, a finding which supports the assumption that chemotherapy down regulates the expression of HGF rather than its degradation

Prognostic relevance of telomerase and Bcl-2 in acute myeloid leukemia

Prognosis of AML patients is influenced by both clinical and genetic markers. As therapy and supportive care improves, the intrinsic biologic characteristics of the patient's leukemia become the dominant factor in determining prognosis. Some reports suggest that telomerase and Bcl-2 may have a prognostic relevance in AML. The objective of this study was to evaluate the prognostic relevance of Telomerase activity and serum level of Bcl-2 in correlation with the disease outcome of AML patients. The study included 63 newly diagnosed cases of acute myeloid leukemia who presented to the Medical Oncology department at the National Cancer Institute, Cairo University. All patients were adults below the age of 60 years. Blood samples were taken from all patients to assess telomerase activity and Bcl-2 levels prior to the administration of anti-neoplastic treatment as well as from ten healthy controls. Assessment of telomerase activity was done using PCR-ELISA technique and evaluation of Bcl-2 serum level was done using ELISA. Patients were followed up for 3 years. Assessment of prognostic factors in the present study included three main independent parameters: cytogenetics abnormalities [20 cases], immunophenotyping [63 cases] and hyperleukocytosis [63 cases]. Patients were also grouped according to the presence of independent prognostic factors into a poor prognosis group and a non-poor prognosis group. Using this classification half the patients [29 patients: 46%] were categorized in the poor prognosis group. Thirty six percent of patients had hyperleukocytosis [TLC >/= 100,000/micro l], 38% expressed unfavorable immunophenotypic markers [CD34 positivity and/or biphenotypic leukemia's markers], while 3 patients had a poor karyotypic profile [11q23, t [9; 22], del 5q]. The complete remission rate was 57% and the overall median time to CR was 31 days. The 2-year and 3-year overall survival rates were 32.5% and 23.5% respectively. While the 2 and 3-year disease-free survival rates were 21.6% and 18% respectively. We found no correlation between the presence of adverse prognostic factors and time to CR, overall survival or disease-tree survival rates. However, patients in the poor prognosis group showed an inferior 2-year disease-free survival [12% versus 34%; p=0.02]. The median level of telomerase activity for AML patients was 0.40 U [0.38 to 0.56]. We found a significant negative correlation between CR rate and telomerase activity [p=0.019] but not with time to CR, or the 2-year overall survival. Lower levels of telomerase activity were associated with a significantly better disease free survival at 1 year when compared to higher levels [34% versus 10%; p=0.012]. There was also a highly significant correlation between telomerase activity and the poor prognosis group [p=0.0001]. The median serum level of Bcl-2 for patients in the present study was 204 U/mL. Bcl-2 levels correlated negatively with CR rate [p=0.06]; but did not correlate with time to CR. Bcl-2 levels less than 200 U/mL were associated with better 2-year overall survival and 1-year disease-free survival [p=0.07 and p=0.005]. There was a highly significant correlation between Bcl-2 and prognosis group [p=0.0001], the presence of hyperleukocytosis [p=0.0001] and CD 34 positivity [p= 0.011]. Patients with more than one poor prognostic criterion had a tendency for lower overall survival rate at 1 year [24% versus 42%; p=0.094] and significantly lower disease-free survival rate at 1 year [24% versus 50%; p=0.036]. There was a significant correlation between telomerase activity levels and Bcl-2 level in the serum of AML patients [r=0.623, p< 0.0001]. Telomerase activity levels and Bcl-2 levels correlate significantly with disease-free survival in AML patients. Further studies would be needed to establish their role in the prognosis of this disease and to determine if an association between telomerase activity and the anti-apoptotic pathway exists

Expression of P-glycoprotein and BCL

Drug resistance is considered to be the major cause of failure of anticancer therapy in patients with malignancy negative regulator of apoptosis such as BCL 2 have also been considered. The aim of this work is to evaluate P-gp and BCL 2 expression level and its significant prognostic value in acute lymphoblastic leukemia [ALL] amid disseminated non-Hodgkin lymphoma [D-NHL]. Peripheral blood mononuclear cells from newly diagnosed cases [30 ALL, 30 D-NHL] and 20 healthy control subjects were collected. lmmunohistochemical staining of mononuclear cells using BCL 2, P-glycoprotein [P-gp] monoclonal antibodies. A Statistically significant increase of P-gp and BCL 2 expression on malignant cells was found in ALL and D-NHL cases compared to the normal control. No statistical significant difference in BCL 2 or P-gp expression in different immunopenotypic patterns, different FAB subtypes of ALL cases or different pathological types or grades of D-NHL cases. A significant statistical correlation was found between the level of BCL 2 and P-gp expression and decreased disease free survival [DFS] and over all survival [OS] in NHL and ALL cases. Assessment of multi drug resistance mechanisms based on both P-gp and BCL 2 expression level is suggested to be an important predictor of treatment outcome in ALL and D-NHL cases

Significance of telomerase activity, C-erbB2, malondialdhyde and NO as biological markers in urine of bladder cancer patients

At the National Cancer Institute [NCI], Cairo, Egypt, bladder neoplasm constitutes 30% of all cancers. Evaluation of urinary markers may hold a promising method for detection of bladder neoplasms with higher sensitivity and specificity, for follow-up in order to regulate the interval of cystoscopic examination, reduce the burden and discomfort of patients amid enhance the opportunities to excise the tumor preceding muscular invasion. The present study aims to evaluate the possible diagnostic role of telomerase activity, C-erbB2, malondialdehyde and NO in the urine of bladder cancer patients. Eighty urine samples were taken from 3 groups of individuals; 1] Ten healthy age matched control subjects, 2] Twenty schistsoma haematobium infested patients and 3] Fifty pre-operative bladder cancer patients. Urine samples [50mL] were collected and subjected to the assay of telomerase activity in urine [TAU], it was measured by PCR-ELISA technique using the telomerase repeat amplification protocol [TRAP], malondialdehyde and nitric oxide were determined spectrophotometrically and C-erbB2 was measured by ELISA technique. TAU was increased in 72% of bladder cancer cases, it was normal in bilhazial non malignant group compared to controls. Its increase in bladder cancer patients with bilharzial infection was statistically insignificant compared to non bilharzial cancer group. TAU of bladder cancer patients were increased in ascending manner with grades of the tumor [GI = 62.5 +/- 16.7, GII = 66.64 +/- 9.37 and GIII = 163 +/- 51]. Malondialdehyde level was increased in bladder cancer patients with bilharzial infestation than those without bilharziasis, but the difference was statistically insignificant. C-erbB2 expression was increased in 27% of bladder cancer patients; while no single case of the bilharzial group showed positive C-erbB2 expression. As regard the stage of tumor NO level in bladder cancer patients showed statistical significant difference between stage I. II amid stage III [p = 0.04]. There was only a statistically significant positive correlation between telomerase and C-erbB2 in bladder cancer patients [r = 0.456 and p = 0.005]. The study of telomerase activity in the urine of bladder cancer cases may be used as an indicator for early detection of this disease. Further studies should be done to evaluate the possibility of using telomerase as one of the most important tumor markers

Expression of human recombinase activating gene 1 and recombinase activating gene 2 [rag 1 and rag 2] in acute lymphoblastic leukemia

The process of V[D]J recombination is limited and controlled by the enzymatic activity of cytoplasmic proteins called recombinase which are the products of the activating two genes called Recombinase Activating Gene 1 and Recombinase Activating Gene 2 [RAG1 and RAG 2]. Both genes are expressed in immature B and T lymphocytes and activated V[D]J rearrangement in Ig and TCR genes which are directed in cis by recombination signals sequences [RSSs]. Also, they show variable expression in lymphoid malignancies of both B and T-cell types. This study aimed to focus on the role of the two cytoplasmic proteins [RAG1 and RAG2] in the developing of both B and T ALL, correlation to the stages of differentiation and also, their possible prognostic significance. This study included 40 newly diagnosed acute lymphoblastic leukemic patients [ALL] their age ranged from 1-10 years. Assessment of RAG1 and RAG2 expression by reverse transcriptase polymerase chain reaction [RT-PCR] was done using peripheral blood sample. RAG1 and RAG2 positive expression was higher in frequency in pediatric ALL cases compared to control group, the difference was statistically significant [P<0.01]. Among the RAG1 and RAG2 positive ALL cases 25% were pro-B-ALL and 25% CALL and 31.2% pre-B-ALL and 18.8% were early T-ALL. On the other hand RAG1 and RAG2 negative ALL cases showed higher frequency of CALL phenotype [83.3%] while pro-B ALL, pre-B ALL and early-T ALL were 4.2% and 8.3% and 4.2% respectively. The RAG1 and RAG2 initially positive ALL cases studied had poor prognosis, where 37.5% relapsed and 25%, while 37.5 were in continuous complete remission. However among the RAG1 and RAG2 negative ALL cases studied, 91.7% had good prognosis with complete remission, while only one patient [4.2%] relapsed and one died [4.2%]. A statistical significant association between RAG1 and RAG 2 positive expression and poor prognosis was noticed. RAG1 and RAG2 could be used as prognostic marker in lymphoid malignancies and its sub-classification

2'-Deoxycytidine as a potential biomarker for detection of hepatocellular carcinoma

Background: 2'-deoxycytidine [Dcyd] is one of four major nucleosides found in the different normal body fluids due to dissolution of dead cells, and is increase in the presence of malignancy. Previous studies proved that it can be used as a marker for bladder cancer and acute lymphoblastic leukemia. The aim of this study is to assess 2'Dcyd as a possible biological marker in hepatocellular carcinoma [HCC]

Methods: four groups were evaluated for the level 2'-Dcyd as well as alpha-fetoprotein [AFP]; a control group [n = 20], 20 cases of chronic liver diseases [CLD], 20 cases of hepatitis C [HCV] 60 cases of HCC

Results: in the patients with HCC, 2'-Dcyd serum level was 8-fold higher than normal level. It was 3-fold higher in HCV group. A mild increase was noted in patients with chronic liver diseases. Levels >/= 0.14 of 2'-Dcyd had a sensitivity of 93% and specificity of 90% for diagnosis of HCC. It also recorded a sensitivity and specificity of 90% for diagnosis of HCV

Conclusions: for diagnosis of HCC, 2'-Dcyd is no better than AFP, as it is elevated in viral hepatitis C. A combination of AFP and 2'-Dcyd could provide broader information in diagnosis and treatment decision