Procaine Induces Epigenetic Changes in HCT116 Colon Cancer Cells.

Colon cancer is the third most commonly diagnosed cancer in the world, and it is the major cause of morbidity and mortality throughout the world. The present study aimed at treating colon cancer cell line (HCT116) with different chemotherapeutic drug/drug combinations (procaine, vorinostat "SAHA," sodium phenylbutyrate, erlotinib, and carboplatin). Two different final concentrations were applied: 3 µM and 5 µM. Trypan blue test was performed to assess the viability of the cell before and after being treated with the drugs. The data obtained showed that there was a significant decrease in the viability of cells after applying the chemotherapeutic drugs/drug combinations. Also, DNA fragmentation assay was carried out to study the effect of these drugs on the activation of apoptosis-mediated DNA degradation process. The results indicated that all the drugs/drug combinations had a severe effect on inducing DNA fragmentation. Global DNA methylation quantification was performed to identify the role of these drugs individually or in combination in hypo- or hypermethylating the CpG dinucleotide all over the genome of the HCT116 colon cancer cell line. Data obtained indicated that different combinations had different effects in reducing or increasing the level of methylation, which might indicate the effectiveness of combining drugs in treating colon cancer cells.

Molecular genotyping and frequencies of A1, A2, B, O1 and O2 alleles of the ABO blood group system in a Kuwaiti population.

Expression of the highly polymorphic ABO gene cluster is commonly investigated for blood transfusion and analysis, but little information is available for Middle Eastern populations. This study determined the major ABO allele frequency in a Kuwaiti Arab cohort using a multiplex PCR-RFLP technique; 355 unrelated blood donors of phenotype A1 (46), A2 (31), A1B (6), A2B (4), B (97) and O (171) were genotyped. DNA fragments of 252 (251 for O1) and 843 (842 for A2) bp spanning the two major exons, 6 and 7, of the ABO gene were amplified and digested with HpaII and KpnI. Thirteen different genotypes could be identified when combining the A1, A2, B, O1 and O2 alleles from the digestion patterns: 1 A1 A1 (0.28%), 6 A1 A2 (1.69%), 38 A1 O1 (10.71%), 1 A1 O2 (0.28%), 1 A2 A2 (0.28%), 30 A2 O1 (8.45%), 6 A1 B (1.69%), 4 A2 B (1.13%), 12 BB (3.38%), 79 BO1 (22.25%), 6 BO2 (1.69%), 167 O1 O1 (47.04%) and 4 O1 O2 (1.13%). Two of the combinations (A2 O2, O2 O2) were not found. All genotypes determined were consistent with the serotypes. The frequencies of the five alleles in the Kuwaiti sample population were ABO*A1 = 0.0746, ABO*A2 = 0.0592, ABO*B = 0.1676, ABO*O1 = 0.6831 and ABO*O2 = 0.0155. These results are discussed with reference to gene frequencies reported for other ethnic groups.

ABO blood group in Kuwaitis: detailed allele frequency distribution and identification of novel alleles.

BACKGROUND: The ABO blood group is clinically the most important blood group system and can now be genotyped easily by DNA-based methods without family studies. STUDY DESIGN AND METHODS: Samples (n = 166) from a Kuwaiti population were phenotyped by standard serologic techniques for the ABO blood group and genotyped for the ABO locus by an established multiplex polymerase chain reaction protocol followed by single-strand conformation polymorphism (SSCP) analysis. Nonstandard SSCP patterns were investigated by DNA sequencing of exons 6 and 7 and, if necessary intron 6. RESULTS: Standard SSCP patterns identified six classical alleles in this population: A101 (0.1115), A102 (0.0181), A201 (0.0301), B101 (0.1627), O101 (0.3103), and O201 (0.2500). One A, 1 B, and 8 O variant alleles were identified (total frequency, 0.1175). All variant alleles were each present in one or two chromosomes (< or =0.0060) in our samples except O109 (0.0813). Three of these 10 variant alleles were novel alleles defined by newly identified single-nucleotide polymorphisms in exon 7 (527G>A, 687C>T, and 1116G>A). One new base substitution result in amino acid change. CONCLUSIONS: This is the first study reporting the detailed distribution of ABO alleles and genotypes in Kuwaitis. Sixteen alleles were identified, including 3 novel alleles.