Anticancer Properties of 3-Dietoxyphosphorylfuroquinoline-4,9-dione.

Herein, the antitumor activity of a novel synthetic analog with 5,8-quinolinedione scaffold, diethyl (2-(2-chlorophenyl)-4,9-dioxo-4,9-dihydrofuro [3,2-g]quinolin-3-yl)phosphonate (AJ-418) was investigated on two breast cancer cell lines. This analog was selected from a small library of synthetic quinolinediones on the basis of its strong antiproliferative activity against MCF-7 and MDA-MB-231 cells and 4-5-fold lower cytotoxicity towards healthy MCF-10A cells. The morphology of MCF-7 and MDA-MB-231 cancer cells treated with AJ-418 changed drastically, while non-tumorigenic MCF-10A cells remained unaffected. In MCF-7 cells, after 24 h incubation, the increased number of apoptotic cells coincided with a decrease in proliferation and cell viability. The 24 h treatment of MDA-MB-231 cells with the tested compound reduced their cell viability and proliferation rate; however, a significant pro-apoptotic effect was visible only after longer incubation times (48 h and 72 h). Then, the maximum tolerated dose (MTD) of compound AJ-418 in C3H mice after subcutaneous administration was determined to be 160 mg/kg, showing that this analog was well tolerated and can be further evaluated to assess its potential therapeutic effect in tumor-bearing mice.

Prognostic Significance of HMGA1 Expression in Lung Cancer Based on Bioinformatics Analysis.

High-mobility group protein 1 (HMGA1) participates in the processes of DNA transcription, replication, recombination, and repair. The HMGA1 gene is expressed abundantly during embryogenesis and is reactivated during carcinogenesis. HMGA1 gene expression has been associated with a high degree of malignancy, metastatic tendency, and poor survival in breast, colon, ovary, and pancreatic cancers. However, its prognostic significance in lung cancer remains unclear. Using publicly available data, HMGA1 was shown to be overexpressed in both small and non-small lung tumors, with higher expression compared to both the adjacent non-malignant lung tissues and non-tumor lung tissues of healthy individuals. Elevated HMGA1 expression could result from lowered HMGA1 methylation and was connected with some clinicopathological features like sex, age, and stage of the disease. The high HMGA1 expression level was connected with shorter overall and first progression survival time among lung adenocarcinoma patients, but not lung squamous cell carcinoma patients. HMGA1 could interact with proteins involved in cellular senescence and cell cycle control (TP53, RB1, RPS6KB1, and CDK1), transcription regulation (EP400 and HMGA2), chromatin assembly and remodeling (LMNB1), and cholesterol and isoprene biosynthesis (HMGCR and INSIG1). Taken together, HMGA1 overexpression could be an essential element of lung carcinogenesis and a prognostic feature in lung cancer.

Synthesis of 2,2,6-Trisubstituted 5-Methylidene-tetrahydropyran-4-ones with Anticancer Activity.

In our continuous search for new, relatively simple 2-alkylidene-1-oxoheterocycles as promising anticancer drug candidates, herein we report an efficient synthesis of 2,2,6-trisubstituted 5-methylidenetetrahydropyran-4-ones. These compounds were obtained in a four step reaction sequence, in which starting diethyl 2-oxopropylphosphonate was transformed into 2,2-disubstituted 5-diethoxyphosphoryldihydropyran-4-ones, followed by Michael addition of various Grignard reagents and Horner-Wadsworth-Emmons reaction of the obtained adducts with formaldehyde. Stereochemistry of the intermediate Michael adducts is also discussed. Final 5-methylidenetetrahydropyran-4-ones were tested for their possible antiproliferative effect against three cancer cell lines and 6-isopropyl-2,2-dimethyl-5-methylidenetetrahydropyran-4-one (11c), which showed very high cytotoxic activity against HL-60 human leukemia cells and was three times more active than known anticancer drug carboplatin, was selected for further biological evaluation, in order to disclose its mechanism of action. The obtained results indicated that 11c induced apoptosis in HL-60 cells and caused the arrest of the cell cycle in the G2/M phase, which may suggest its cytotoxic and cytostatic activity.

Decreased MMP1 gene expression in acute myeloid leukaemia.

Acute myeloid leukaemia (AML) is a heterogeneous disorder of haematopoietic stem cells or progenitor cells. Metalloproteinases (MMPs) are proteolytic enzymes whose activity is increased in different types of solid tumours. These enzymes regulate many processes associated with tumour progression. In haematological malignancy, the role of MMPs seems to be underestimated, and only metalloproteinase 2 (MMP2) and metalloproteinase 9 (MMP9) have been widely examined so far. In this work, differences in metalloproteinase 1 (MMP1) gene expression between patients with AML and healthy individuals were assessed. The relative expression level of the MMP1 gene was obtained by a real time PCR method preceded by reverse transcription. The relative level of MMP1 gene expression in patients with AML was decreased when compared to that of the control group. The role of MMP1 in AML could be different from that in solid tumours. Decreased MMP1 gene expression in AML similar to that of MMP9, shows a greater role for MMP1 in normal haematopoiesis than in the development of leukaemic cells.

Synthesis of 3-Methylidene-1-tosyl-2,3-dihydroquinolin-4(1H)-ones as Potent Cytotoxic Agents.

An efficient synthetic strategy to 3-methylidene-2,3-dihydroquinolin-4(1H)-ones variously substituted in position 2 has been developed. The title compounds were synthesized in the reaction sequence involving reaction of diethyl methylphosphonate with methyl 2-(tosylamino)benzoate, condensation of thus formed diethyl 2-oxo-2-(2-N-tosylphenyl)ethylphosphonate with various aldehydes followed by successful application of the obtained 3-(diethoxyphosphoryl)-1,2-dihydroquinolin-4-ols as Horner-Wadsworth-Emmons reagents for the olefination of formaldehyde. Also, enantioselective approach to the target compounds has been evaluated using 3-dimenthoxyphosphoryl group as a chiral auxiliary. Single X-ray crystal analysis of (2S)-3-(dimenthoxyphosphoryl)-2-phenyl-1-tosyldihydroquinolin-4-ol revealed the presence of strong resonance-assisted hydrogen bond (RAHB). The obtained 3-methylidene-2,3-dihydroquinolin-4(1H)-ones were then tested for their cytotoxic activity against two leukemia cell lines NALM-6 and HL-60 and a breast cancer MCF-7 cell line. All compounds showed very high cytotoxic activity with the IC50 values mostly below 1 µm in all three cancer cell lines. The selected analogs were also tested on human umbilical vein endothelial cells (HUVEC) and on human mammary gland/breast cells (MCF-10A) to evaluate their influence on normal cells. Since one of the most serious problems in cancer chemotherapy is the development of drug resistance, the mRNA levels and activity of ABCB1 transporter considered to be the most important factor engaged in drug resistance, were evaluated in MCF-7 cells treated with two selected analogs. Both compounds were strong ABCB1 transporter inhibitors that could prevent efflux of anticancer drugs from cancer cells.

Association of ABCB1 T-129C polymorphism and multiple myeloma risk in Polish population.

The possible interaction between gene polymorphism and cancer risk development is a very interesting issue. The genetic variants of the ATP-binding cassette superfamily B member 1 (ABCB1) are known to be involved in developing cancer risk and individual differences in chemotherapeutic response. Polymorphisms may affect the reduction of the activity and/or expression of important protective cellular proteins. The increased exposure to toxic compounds, including carcinogens is associated with an increased risk of developing cancers. The present study was aimed to evaluate the possible effect of ABCB1 T-129C single nucleotide polymorphism in risk of cancer development in Polish patients diagnosed with multiple myeloma. 91 multiple myeloma patients and 94 healthy controls were enrolled in this case-control study. The ABCB1 T-129C genotypes were determined by polymerase chain reaction-restriction fragment length polymorphism method (PCR-RFLP). The distribution of particular genotypes between multiple myeloma patients and controls group was not significantly different for T-129C SNP (p = 0.4297). The studied polymorphism does not seem to affect the increased risk of multiple myeloma development.

The Influence of C3435T Polymorphism of the ABCB1 Gene on Genetic Susceptibility to Depression and Treatment Response in Polish Population - Preliminary Report.

BACKGROUND: Despite the high prevalence of depression, the mechanism of the origin of this disease as well as the causes of resistance to therapy in some patients are still not fully understood. Increasingly, the possible role of genetic factors is considered. One of them is polymorphisms in the ABCB1 (MDR1) gene which encodes P-glycoprotein, responsible for the transport of xenobiotics, including antidepressant drugs, through the blood-brain barrier. METHODS: C3435T was evaluated in 90 patients with recurrent depressive disorders (rDD). Genotyping was performed using a polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP). RESULTS: The obtained results indicate that the TT genotype occurred more frequently among patients with rDD than in healthy volunteers (p=0.0441). Also, at least one C allele was present significantly less frequent in the study group than in healthy individuals (p=0.0300). The severity of depressive symptoms was higher among patient with the CC genotype in comparison with the other genotypes (p=0.0106) but treatment response to antidepressants was better in this group than among patients with CT or TT genotypes (p=0.0301). Likewise, patients with the T allele have a significantly lower severity of symptoms (p=0.0026) and decreased therapy effectiveness (p=0.0142) than C allele carriers. CONCLUSIONS: This study suggests that C3435T polymorphisms in the ABCB1 gene are strongly associated with a predisposition to depression development, the severity of depressive symptoms and the effectiveness of therapy with using different groups of antidepressant agents.

Arsenic trioxide downregulates cancer procoagulant activity in MCF-7 and WM-115 cell lines in vitro.

THE AIM OF THE STUDY: To analyze human breast cancer cell line MCF-7 and human malignant melanoma cell line WM-115 in order to characterize the cellular expression of CP and to evaluate whether ATO may affect this activity, as well as the viability of the cells. MATERIAL AND METHODS: The inhibitory effect of arsenic trioxide on the proliferation of MCF-7 and WM-115 cells were measured with MTT test. The activity of cancer procoagulant after ATO exposure was determined by a specific three-stage chromogenic assay. RESULTS: ATO decreased the CP activity in a dose- and time-dependent manner in MCF-7 cells with no effect on cell proliferation at the same time. However, it affected the CP activity of WM-115 cells in a different way. Reduction in CP activity was followed by an increase after 48 h incubation. The cells viability results showed dose-and time-correlated response within high arsenic concentrations. CONCLUSIONS: Arsenic trioxide downregulates the CP expression in human breast cancer and melanoma cells.

Fibrinogen, bFGF and VEGF levels during antibiotic therapy in gynecologic cancer: a preliminary report.

The role of angiogenesis in the development of neoplasia has been identified and characterized. However, antiangiogenic therapeutic intervention still requires more evidence to become recognized and successful. The aim of this study was to evaluate levels of selected proangiogenic factors, such as fibrinogen, vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) in serum of patients with the gynecologic cancer on the first, third and sixth day of antibiotic therapy, routinely administered as a perioperative treatment. In addition, serum concentrations of gamma-gamma dimers and alpha-polymers of cross-linked fibrin structure and the degree of bFGF binding with the fibrin network were investigated. Immunohistochemistry staining of the excised tumor tissue was also performed. We observed higher levels of bFGF, VEGF, as well as fibrinogen in patients with gynecologic malignancy, as compared to healthy women. In cancer patients, the concentration of alpha-polymers and gamma-gamma dimers of fibrin network increased. Further only gamma-gamma dimers fraction of fibrin was found to bind to bFGF. Immunohistochemical analysis indicated the presence of bFGF in an excised tumor tissue. In conclusion, the decrease of proangiogenic bFGF and fibrinogen levels in a clinical trial of gynecologic patients may confirm anti-angiogenic properties of selected antibiotic therapy.

Historical Perspective and Current Trends in Anticancer Drug Development.

Cancer is considered one of the leading causes of death in the 21st century. The intensive search for new anticancer drugs has been actively pursued by chemists and pharmacologists for decades, focusing either on the isolation of compounds with cytotoxic properties from plants or on screening thousands of synthetic molecules. Compounds that could potentially become candidates for new anticancer drugs must have the ability to inhibit proliferation and/or induce apoptosis in cancer cells without causing too much damage to normal cells. Some anticancer compounds were discovered by accident, others as a result of long-term research. In this review, we have presented a brief history of the development of the most important groups of anticancer drugs, pointing to the fact that they all have many side effects.

Effect of phosphorohydrazone derivatives of chromone on fibrin polymerization in the presence of bFGF.

The effect of ten phosphorohydrazone derivatives of chromone on the fibrin formation process was evaluated in the presence of basic fibroblast growth factor (bFGF), which plays an important role in tumor angiogenesis. The relationship between the chemical properties of the derivatives and the resulting fibrin structure was also examined. The structure of fibrin was analyzed by UV spectrophotometry and the degree of bFGF binding to fibrin was estimated by Western blotting. The measurements were taken at 3 different pH values: 6.64, 7.60 and 8.40. Three of the analyzed phosphorohydrazone derivatives (compounds 2, 7 and 10) demonstrated the most significant influence on reduction of polymerization at the studied pH values.

Quantitative analysis of FJ 194940.1 gene expression in colon cancer and its association with clinicopathological parameters.

AIM OF THE STUDY: The FJ 194940.1 gene is located on chromosome 1 and consists of 6 exons and 5 introns. The gene undergoes alternative splicing and its isoforms appear during cancer development. Evidence suggests that expression of FJ 194940.1 splice variants relate to colorectal cancer progression. This paper discusses the quantitative analysis of the exon V expression level of FJ 194940.1 in colon cancer. The aim of the study is to carry out quantitative analysis by real-time PCR in a series of 102 colon cancer samples that had previously shown presence of exon V expression. To compare the exon V expression level with certain histological parameters and clinical staging of the neoplasm in order to assess its potential role as a prognostic factor in colon cancer. MATERIAL AND METHODS: Tissue specimens of colorectal cancer were obtained from the Oncological Centre of Lodz, Poland.Total RNA isolation was performed in accordance with the protocol enclosed in the Total RNA Prep Plus Minicolumn Kit (A&A Biotechnology, Poland). Reverse Transcriptase-PCR reaction was carried out using Enhanced Avian HSRT-PCR Kit, Sigma, according to the manufacturer's protocol.. Presence of cDNA in each sample was checked by PCR amplification of ß-actin. Only samples showing the PCR product of this housekeeping gene were included in further tests. The amount of FJ 194940.1 transcript containing exon V was analysed by means of real-time PCR. RESULTS: Exon V expression level is not significantly related to any clinicopathological features in colon cancer. However, there was a tendency towards a lower exon V expression level in a group of cases where vessel invasion was present (p = 0.0697). Additionally, the risk of death in patients with a low exon V expression level was more than two times higher when compared to patients with a high exon V expression level. CONCLUSIONS: FJ 194940.1 gene expression correlates with cancer progression independently of analysed clinicopathological parameters.

Investigation of ABCB1 1236 and 2677 SNPs in patients with peptic ulcer.

OBJECTIVE: P-gp, encoded by ABCB1 gene, is an ATP-binding membrane pump, which exports substrates from the cell including drugs and xenobiotics. Changes in the function of P-gp as a result of polymorphism could have an impact in some diseases' risks and treatment outcomes. The aim of the study was to determine the significance of the ABCB1 gene SNPs: 1236 and 2677 for peptic ulcer risk and development of Helicobacter pylori infection in peptic ulcer patients. MATERIAL AND METHODS: One hundred and ninety-five biopsy specimens obtained from peptic ulcer patients (investigated group) were genotyped using sequencing for common SNPs of ABCB1: 1236 and 2677. Genotyping data were compared with the results from healthy subjects (control) and with the presence of H. pylori infection, which was estimated by urease test. RESULTS: No statistically significant difference in frequency of genotypes and alleles for the SNPs were found between the investigated group and the control. However, in the peptic ulcer patients, mutant TT homozygotes and those who carried at least one allele T for the polymorphisms 1236 and 2677 were observed more frequently than the control group. In the peptic ulcer group, there were no significant dependences between the presence of H. pylori infection and the investigated polymorphisms other than more frequent occurrence of TT 1236 homozygous in the group of infected women (p = 0.0298). CONCLUSIONS: The TT genotype and the mutated allele T for the polymorphisms 1236 and 2677 could increase peptic ulcer risk. ABCB1 1236 polymorphism may also be associated with an increased likelihood of H. pylori infection development, especially in women.

Changes in the expression of membrane type-matrix metalloproteinases genes (MMP14, MMP15, MMP16, MMP24) during treatment and their potential impact on the survival of patients with non-small cell lung cancer (NSCLC).

The analysis concerned the comparison of the expression of membrane type matrix metalloproteinases genes in the blood and tissue of NSCLC patients during the course of the disease and comparison to the control group. Blood and neoplastic tissue taken from 45 patients diagnosed with non-small cell lung cancer was a research material. The expression level of MMP14, MMP15, MMP16 and MMP24 was evaluated by qPCR and the results were compared with controls. The expression of MMP14 and MMP24 before tumor removal surgery and 100 days after was lower than in the control group. Interestingly, one year after surgery the levels of expression of these genes were identical to those in the control group. This suggests that the expression of metalloproteinase genes changes in the course of cancer and that effective treatment results in the normalization of gene expression. Lower expression of MMP15 in the blood of patients with more advanced cancer disease was observed, confirming the suppressive nature of changes in the blood. It has also been demonstrated that higher expression of MMP14 and MMP15 in the tissue is associated with more advanced stage of disease development or more invasive nature of the lesion. There is a noticeable increase of expression level in the environment surrounding the tumor, while a lower can be observed in the blood. This may indicate that changes in the expression of metalloproteinases in cancer are much more complex than merely the tumor tissue, which may also account for the inadequacies of metalloproteinase inhibitors.

Importance of Altered Gene Expression of Metalloproteinases 2, 9, and 16 in Acute Myeloid Leukemia: Preliminary Study.

Acute myeloid leukemia is a group of hematological neoplasms characterized by a heterogeneous course and high mortality. The important factor in the neoplastic process is metalloproteinases, proteolytic enzymes capable of degrading various components of the extracellular matrix, which take an active part in modifying the functioning of the cell, including transformation to cancer cell. They interact with numerous signaling pathways responsible for the process of cell growth, proliferation, or apoptosis. In the present study, changes in the expression of MMP2, MMP9, and MMP16 genes between patients with AML and people without cancer were examined. The impact of cytogenetic changes in neoplastic cells on the expression level of MMP2, MMP9, and MMP16 was also assessed, as well as the impact of the altered expression on the effectiveness of the first cycle of remission-inducing therapy. To evaluate the expression of all studied genes MMP2, MMP9, and MMP16, SYBR Green-based real-time PCR method was used; the reference gene was GAPDH. For two investigated genes MMP2 and MMP16, the lower expression level was observed in patients with AML when compared to healthy people. The MMP9 gene expression level did not differ between patients with AML and healthy individuals which may indicate a different regulation of gene expression in acute myeloid leukemia. However, no correlation was observed between the genes' expression of all tested metalloproteinases and the result of cytoreductive treatment or the presence of cytogenetic changes. The obtained results show that the expression of MMP2 and MMP16 genes is reduced while the expression of MMP9 is unchanged in patients with acute myeloid leukemia. This may indicate a different regulation of the expression of these genes, and possible disruptions in gene transcription or posttranscriptional mechanisms in the MMP2 and MMP16 genes, however, do not affect the level of MMP9 expression. Obtained results in AML patients are in contrary to various types of solid tumors where increased expression is usually observed.

ABCB1/MDR1 gene polymorphisms as a prognostic factor in colorectal cancer.

OBJECTIVE: To analyse the single-nucleotide polymorphisms (SNPs): ABCB1(1236C>T), ABCB1(2677G>T/A), ABCB1(3435C>T) and haplotypes in the ABCB1/MDR1 gene, which could contribute to genetic risk of colorectal cancer (CRC). Disease association between the ABCB1/MDR1 genotype, allele, haplotype frequencies and histological features, such as TNM classification, localization of primary carcinoma, grade of malignancy, histological type of tumour, lymphoid infiltration and vessel invasion were estimated. In this study, the potential role of SNPs of the ABCB1/MDR1 gene as a prognostic marker for CRC was analysed. MATERIALS AND METHODS: Tumour specimens of 95 patients with CRC were studied. Using automated sequencing or PCR-RFLP method, DNA for three common SNPs of ABCB1/MDR1 was extracted and analysed. The results of genotyping and haplotype analysis with histopathological features, grading and clinical staging of neoplasms were correlated. RESULTS: A statistically significant higher frequency of T(1236) allele in T1/T2 (89.7%), M0 groups (81.6%) and I/II clinical staging (82.7%) in comparison with T3/T4 (68.2%), M1 groups (47.4%) and III/IV clinical staging (65.1%) was detected. Furthermore, multivariate analysis according to Cox's proportional hazard model indicated that the T(1236) allele is a good, independent prognostic factor and the presence of this allele decreases the risk of death in comparison with a group without this allele (HR = 0.26; p = 0.0424). In addition, a statistically significant higher frequency of C(3435) allele and significant differences in the C(3435) allele distribution in N1/N2 group (91.7% and 62.5%, respectively) than N0 group (71.2% and 44.9%, respectively) was found. Each of the eight possible haplotypes was noted in M0 or I/II group and only seven in M1 or III/IV group. Haplotype T(1236)-G(2677)-C(3435) only in less advanced CRC subjects (9.6% in I/II and 9.2% in M0 group) was detected. In addition, significant differences in haplotype distributions between M0 or I/II and M1 or III/IV group were found (p = 0.01 and p = 0.05, respectively). CONCLUSIONS: These results suggest association between T(1236) allele and T(1236)-G(2677)-C(3435) haplotype and less advanced CRC, so these genetic markers may play a role as potentially good prognostic factors. Differences in haplotype distributions and degree of clinical staging may suggest that some other potential SNPs, especially in regulatory region of ABCB1/MDR1 gene, may influence P-glycoprotein function and CRC progression.

New Uracil Analogs with Exocyclic Methylidene Group as Potential Anticancer Agents.

BACKGROUND: Hybrid molecules combining uracil skeleton with methylidene exo-cyclic group were designed in the search for novel anticancer drug candidates. OBJECTIVE: Two series of racemic 5-methylidenedihydrouracils, either 1,3-disubstituted or 1,3,6-trisubstituted were synthesized and tested for their possible cytotoxic activity against two cancer cell lines (HL-60 and MCF-7) and two healthy cell lines (HUVEC and MCF-10A). The most cytotoxic analogs were re-synthesized as pure enantiomers. The analog designated as U-332 [(R)-3-(4-bromophenyl)-1-ethyl-5-methylidene-6-phenyldihydrouracil], which had a very low IC50 value in HL-60 cell line (0.77µM) and was the most selective towards cancer cells was chosen for further experiments on HL-60 cell line, in order to determine the possible mechanism involved in its antineoplastic action. METHODS: Cytotoxic activities of compound was assessed by the MTT assay. In order to explore the mechanism of U-332 activity, we performed quantitative real-time PCR analysis of p53 and p21 genes. Apoptosis, cell proliferation and DNA damage in HL-60 cells were determined using the flow cytometry. The ability of U-332 to determine GADD45ɑ protein level in HL-60 cells incubated with U-332 was analyzed by ELISA test. RESULTS: U-332 was shown to generate excessive DNA damage (70% of the cell population), leading to p53 activation, resulting in p21 down-regulation and a significant increase of GADD45α protein, responsible for the cell cycle arrest in G2/M phase. CONCLUSION: U-332 can be used as a potential lead compound in the further development of novel uracil analogs as anticancer agents.

The impact of ABCB1 gene polymorphism and its expression on non-small-cell lung cancer development, progression and therapy - preliminary report.

The ABCB1 gene belongs to ATP binding cassette (ABC) transporter genes that has been previously implicated in cancer progression and drug response. This study aimed to evaluate the association between the SNP 3435 and the expression of the ABCB1 gene in lung cancer patients in the Polish population in comparison to clinicopathological parameters and treatment. 150 RNA and 47 DNA samples were isolated from 49 lung cancer cases including both tissue samples and blood taken from the same patients at three time points: diagnosis, 100 days and one year after the surgical intervention. Qualitative and real-time PCR analysis of expression were done, also genotyping by PCR-RFLP. Mutant homozygous TT and allele T are present statistically significantly more frequently in the group of patients with lung cancer. There is no difference with expression level in lung cancer tissue and blood sample taken from the same patients before surgical treatment. On the basis of blood samples analysis it was observed that the expression level of ABCB1 mRNA was growing in time. Higher levels were marked after 100 days and one year after the surgical intervention. The complementary pharmacological treatment induced higher expression levels of ABCB1. The presented data suggest an important role of ABCB1 in lung cancer, the increasing level of ABCB1 mRNA which can be connected with induction of multidrug resistance mechanism is also significant, that observation must be confirmed in further analysis.

CD38 gene polymorphisms contribute to genetic susceptibility to B-cell chronic lymphocytic leukemia: evidence from two case-control studies in Polish Caucasians.

Given the recent findings on the importance of CD38 signaling in the pathogenesis of B-cell chronic lymphocytic leukemia (B-CLL), we hypothesized that single nucleotide polymorphisms (SNP) in the CD38 gene may be related to B-CLL risk. We evaluated two potentially functional CD38 SNPs, intronic rs6449182 (184C>G) and missense rs1800561 (418C>T, Arg140Trp) in two hospital-based case-control studies (study A and validation study B). Genotyping was done using PCR-based assays in a total of 460 Polish Caucasian patients with B-CLL and 503 age-matched and gender-matched controls. We found that frequencies of both variant alleles (rs6449182 G and rs1800561 T) were significantly higher in B-CLL. In study A, logistic regression analysis revealed an association between B-CLL and genotypes: rs6449182 CG [odds ratio (OR), 3.57; 95% confidence interval (95% CI), 2.4-5.3], rs6449182 GG (OR, 5.2; 95% CI, 2.36-11.5), and rs1800561 CT (OR, 6.72; 95% CI, 1.5-30.1), although no homozygous rs1800561 TT genotype was detected in either study. These results were confirmed in study B, which showed an association between B-CLL and genotypes rs6449182 CG (OR, 4.00; 95% CI, 2.7-6.0), rs6449182 GG (OR, 12.84; 95% CI, 4.3-38.7), and rs1800561 CT (OR, 10.12; 95% CI, 1.3-81.6), and in the combined analysis of both studies. We also observed that rs6449182 G carriers had more advanced clinical stage (P=0.002) and tended to be younger at diagnosis (P=0.056). Furthermore, we found higher CD38 transcript levels and higher proportions of CD38-positive cells in carriers of rs6449182 G and rs1800561 T alleles (P<0.05 for all comparisons). In conclusion, our data show that CD38 SNPs may affect CD38 expression and contribute to the increased risk of B-CLL carcinogenesis.

ABCB1 gene polymorphisms and haplotype analysis in colorectal cancer.

PURPOSE: The aim of this study was to determine the significance of three most common single-nucleotide polymorphisms (SNPs) of ABCB1 gene in the development of colorectal cancer and to estimate the influence of these SNPs to surviving patients' treatment combination adjuvant therapy 5-fluorouracil/leucovorin. Haplotype structure of ABCB1 was analysed, and degree of linkage disequilibrium (LD) between SNPs of ABCB1 was estimated. MATERIALS AND METHODS: Tumour specimens of 95 patients with colorectal cancer and blood samples of 95 healthy cases were studied. Genotyping of ABCB1 gene was performed by automated sequencing or polymerase chain reaction-restriction fragment length polymorphism method. Comparison of frequencies of alleles/genotypes/haplotypes between the studied group (colorectal cancer samples) and the control group (blood samples) were analysed. These results were correlated with the surviving patients after treatment of adjuvant chemotherapy. RESULTS: Significant differences in ABCB1 (1236C>T) (p = 0.00043) and ABCB1 (2677G>T/A) (p = 0.04) genotype distribution and T(1236) allele distribution (CT(1236) or TT(1236) vs CC(1236); p = 0.0499, OR = 0.55, Fi-Yule coefficient = 0.14) were found. A strong LD between ABCB1 (1236C>T) and ABCB1 (2677G>T/A) SNPs (D' = 0.621, r (2) = 0.318) was detected. All SNPs were located in one haplotype block. There were significant differences in haplotype distributions between colorectal cancer patients and healthy population (p = 0.03). Significant differences in survival probability of colorectal cancer patients' treatment chemotherapy according to allele of ABCB1 (3435C>T) was observed. Survival probability of patients with wild-type C(3435) allele were higher than among patients without this allele (p = 0.04572). CONCLUSIONS: These results suggested that three studied SNPs of ABCB1 were located in one haplotype block. Differences in ABCB1 (1236C>T) and ABCB1 (2677G>T/A) genotypes and T(1236) allele distribution between investigated populations indicate significant impact of these SNPs on risk of development of colorectal cancer. Polymorphism ABCB1 (3435C>T) may be a prediction marker of cancer chemotherapy effectiveness. Differences in haplotype distributions between colorectal cancer patients and healthy population suggested that other potential SNPs, especially in regulatory region of ABCB1 gene, may influence P-glycoprotein expression and function.