Standards of diagnostic and new trends in treatment in patients with acute myeloid leukemia

Acute myeloid leukemia (AML) is the most common cancer of white blood cells in adults. Men over 65 years old are more prone to develop this disease. Symptoms that lead patients to visit the doctor are: high fever, bone pain, weakness and signs of infection. The etiology of AML is not yet fully understood. The predisposing factors for acute myeloid leukemia may include environmental and genetic factors. If left untreated, it can lead to death within a few weeks. Therefore, it is important to quickly identify the disease and to implement appropriate treatment, which will allow to increase the percentage of survival among patients. The basis of AML diagnosis is the presence of more than 20% of blasts in blood or bone marrow smears. The choice of AML treatment depends on prognostic factors: patients’ age and sex and cytogenetic-molecular risk. The treatment regimen for AML is outdated and remains almost unchanged for over 30 years. Understanding the molecular basis of this disease development and pathomechanism allows to search for new effective treatments, often based on targeted therapies. This article presents contemporary standards of AML diagnosis and the latest trends in its treatment

Estimation of sodium/iodide symporter gene expression (NIS) in thyroid cancer by RT-PCR technique (preliminary study)

Gen NIS zlokalizowany na chromosomie 19 koduje białko złożone z 643 aminokwasów. Białko to należy do rodziny białek błonowych tzw. zależnych od Na+ symporterów glukozowych. W prawidłowej tarczycy jest zlokalizowane w błonie podstawnej tyreocytu. Fizjologiczna rola białka NIS polega na współudziale w biosyntezie hormonów tarczycy, umożliwia bowiem nagromadzanie w znacznym stężeniu jonów I- w tyreocycie. Z doniesień literaturowych wynika, że w nowotworach złośliwych tarczycy (głównie pęcherzykowych i brodawkowatych) ekspresja genu NIS może wpływać na skuteczność rutynowo stosowanej terapii jodem radioaktywnym. Celem pracy było określenie ekspresji genu NIS w rakach tarczycy, a także poszukiwanie zależności pomiędzy ekspresją badanego genu a stopniem zaawansowania klinicznego nowotworu oraz odpowiedzią na zastosowaną radioterapię. Materiał do badań stanowiły mrożone skrawki tkankowe pochodzące z raków tarczycy. Metoda którą wykorzystano do badań była oparta na reakcji odwrotnej transkrypcji (RT) wyizolowanego RNA, po której następowała reakcja PCR wykorzystująca startery swoiste dla genu NIS. Na podstawie uzyskanych wyników stwierdzono, że ekspresja genu NIS pojawia się w różnych typach histologicznych raków tarczycy, głównie rakach brodawkowatych. Nie stwierdzono istotnych statystycznie zależności pomiędzy obecnością ekspresji genu NIS, a stopniem zaawansowania klinicznego zbadanych nowotworów wg klasyfikacji TNM. Nie znaleziono istotnych statystycznie zależności pomiędzy ekspresją badanego genu, a płcią i wiekiem pacjenta. Na bazie dotychczasowych wyników nie ustalono związku pomiędzy ekspresją genu NIS, a jodochwytnością zbadanych przypadków. Ustalenie zależności między ekspresją badanego genu, a skutecznością jodoterapii wymaga dalszych badań, z wykorzystaniem metody ilościowej.NIS gene is located on chromosome 19 and encodes 643 amino acid protein. It belongs to membrane Na+ dependent glucose symporter proteins family. In normal thyroid is located in basolateral membrane of thyreocyte. It plays a main role in concentrating of iodine in thyreocyte and thus in thyroid hormones synthesis. It was proved that NIS expression influences effectiveness of radioactive iodine therapy in well-differentiated thyroid cancers. The aim of this study was to estimate the NIS expression and its dependence with gender, age and stage in thyroid papillary and follicular cancers. The frozen sections of tissue were used as a source of tumor RNA. RT-PCR technique was employed for NIS expression analysis. We did not find dependence between the presence of NIS expression in investigated thyroid cancers and stage of disease estimated according to TNM classification. We also did not find dependence between NIS expression and gender or sex of the patients. Our results suggest that there is no dependence between NIS expression and iodine uptake

Isolation and characterization of a copalyl diphosphate synthase gene promoter from Salvia miltiorrhiza

The promoter, 5' UTR, and 34-nt 5' fragments of protein encoding region of the Salvia miltiorrhiza copalyl diphosphate synthase gene were cloned and characterized. No tandem repeats, miRNA binding sites, or CpNpG islands were observed in the promoter, 5' UTR, or protein encoding fragments. The entire isolated promoter and 5' UTR is 2235 bp long and contains repetitions of many cis-active elements, recognized by homologous transcription factors, found in Arabidopsis thaliana and other plant species. A pyrimidine-rich fragment with only 6 non-pyrimidine bases was localized in the 33-nt stretch from nt 2185 to 2217 in the 5' UTR. The observed cis-active sequences are potential binding sites for trans-factors that could regulate spatio-temporal CPS gene expression in response to biotic and abiotic stress conditions. Obtained results are initially verified by in silico and co-expression studies based on A. thaliana microarray data.The quantitative RT-PCR analysis confirmed that the entire 2269-bp copalyl diphosphate synthase gene fragment has the promoter activity.Quantitative RT-PCR analysis was used to study changes in CPS promoter activity occurring in response to the application of four selected biotic and abiotic regulatory factors; auxin, gibberellin, salicylic acid, and high-salt concentration

Human leukocyte antigens HLA DRB1 influence clinical outcome of chronic lymphocytic leukemia

We investigated HLA DRB1 correlations with chronic lymphocytic leukemia (B-CLL) outcome in 90 patients. Neither of the alleles was associated with B-CLL clinical characteristics or mortality. HLA DRB1*01 and HLA DRB1*02-null were associated with shorter overall survival (p=0.007, p=0.002). Our results suggest that HLA-restricted adaptive immunity influences CLL outcome

Overexpression of HMGA1 promotes anoikis resistance and constitutive Akt activation in pancreatic adenocarcinoma cells

HMGA1 proteins are architectural transcription factors that are overexpressed by pancreatic adenocarcinomas. Roles of HMGA1 in mediating the malignant phenotype of this cancer are poorly understood. We tested the hypothesis that overexpression of HMGA1 promotes resistance to anoikis (apoptosis induced by anchorage deprivation) in pancreatic cancer cells. HMGA1 cDNA was stably transfected into MiaPaCa2 human pancreatic adenocarcinoma cells (which have low baseline expression levels of HMGA1). Cells were grown in suspension on PolyHEMA-coated plates and their susceptibility to anoikis was assayed using flow cytometry. Overexpression of HMGA1 was associated with marked reductions in susceptibility to anoikis in concert with increases in Akt phosphorylation (Ser473) and in Akt kinase activity and with reductions in caspase 3 activation. Inhibition of phosphoinositidyl-3 (PI3-K)/Akt pathway with either the small molecule inhibitor LY294002 or dominant-negative Akt resulted in reversal of anoikis resistance induced by HMGA1 overexpression. Further, RNA interference-mediated HMGA1 silencing in MiaPaCa2 and BxPC3 (a human pancreatic adenocarcinoma cell line with high baseline levels of HMGA1 expression) cells resulted in significant increases in susceptibility to anoikis. Our findings suggest HMGA1 promotes anoikis resistance through a PI3-K/Akt-dependent mechanism. Given the putative associations between anoikis resistance and metastatic potential, HMGA1 represents a potential therapeutic target in pancreatic adenocarcinoma

Effect of ABCB1 and ABCC3 Polymorphisms on Osteosarcoma Survival after Chemotherapy: A Pharmacogenetic Study

Background: Standard treatment for osteosarcoma patients consists of a combination of cisplatin, adriamycin, and methotrexate before surgical resection of the primary tumour, followed by postoperative chemotherapy including vincristine and cyclophosphamide. Unfortunately, many patients still relapse or suffer adverse events. We examined whether common germline polymorphisms in chemotherapeutic transporter and metabolic pathway genes of the drugs used in standard osteosarcoma treatment may predict treatment response. Methodology/Principal Findings: In this study we screened 102 osteosarcoma patients for 346 Single Nucleotide Polymorphisms (SNPs) and 2 Copy Number Variants (CNVs) in 24 genes involved in the metabolism or transport of cisplatin, adriamycin, methotrexate, vincristine, and cyclophosphamide. We studied the association of the genotypes with tumour response and overall survival. We found that four SNPs in two ATP-binding cassette genes were significantly associated with overall survival: rs4148416 in ABCC3 (per-allele HR = 8.14, 95%CI = 2.73-20.2, p-value = 5.1×10 -5), and three SNPs in ABCB1, rs4148737 (per-allele HR = 3.66, 95%CI = 1.85-6.11, p-value = 6.9×10 -5), rs1128503 and rs10276036 (r 2 = 1, per-allele HR = 0.24, 95%CI = 0.11-0.47 p-value = 7.9×10 -5). Associations with these SNPs remained statistically significant after correction for multiple testing (all corrected p-values [permutation test] ≤0.03). Conclusions: Our findings suggest that these polymorphisms may affect osteosarcoma treatment efficacy. If these associations are independently validated, these variants could be used as genetic predictors of clinical outcome in the treatment of osteosarcoma, helping in the design of individualized therapyThis work was supported by the AECC (Asociación Española contra el Cáncer), FIS (Fondo de Investigación Sanitaria-Instituto de Salud Carlos III) and the ‘‘Inocente Inocente’’ Foundatio

A Comprehensive Investigation on Common Polymorphisms in the MDR1/ABCB1 Transporter Gene and Susceptibility to Colorectal Cancer

ATP Binding Cassette B1 (ABCB1) is a transporter with a broad substrate specificity involved in the elimination of several carcinogens from the gut. Several polymorphic variants within the ABCB1 gene have been reported as modulators of ABCB1-mediated transport. We investigated the impact of ABCB1 genetic variants on colorectal cancer (CRC) risk. A hybrid tagging/functional approach was performed to select 28 single nucleotide polymorphisms (SNPs) that were genotyped in 1,321 Czech subjects, 699 CRC cases and 622 controls. In addition, six potentially functional SNPs were genotyped in 3,662 German subjects, 1,809 cases and 1,853 controls from the DACHS study. We found that three functional SNPs (rs1202168, rs1045642 and rs868755) were associated with CRC risk in the German population. Carriers of the rs1202168_T and rs868755_T alleles had an increased risk for CRC (Ptrend = 0.016 and 0.029, respectively), while individuals bearing the rs1045642_C allele showed a decreased risk of CRC (Ptrend = 0.022). We sought to replicate the most significant results in an independent case-control study of 3,803 subjects, 2,169 cases and 1,634 controls carried out in the North of Germany. None of the SNPs tested were significantly associated with CRC risk in the replication study. In conclusion, in this study of about 8,800 individuals we show that ABCB1 gene polymorphisms play at best a minor role in the susceptibility to CRC