Effect of Batissa violacea celebensis Martens, 1897, Extract on β-Catenin Gene in Mice (Mus musculus) Induced by Azoxymethane and Dextran Sulfate Sodium using Polymerase Chain Reaction-restriction Fragment Length Polymorphism Method

Abstract

BACKGROUND: At present, the β-catenin gene is a genetic change considered as the most important factor in the initiation and the progress of cancer. Bivalve extract contains steroid compounds that are thought to have an aphrodisiac and anti-inflammatory effect. The triterpenoid component detected in the crude bivalves extract is thought to have antitumor activity. AIM: This study aimed at knowing the effect of ethanol (EtOH) extract, ethyl acetate fraction, and the n-hexane fraction of pokea clam on β-catenin gene mutation in mice induced by azoxymethane (AOM) and dextran sulfate sodium (DSS) using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. METHODS: The testing of the effect of pokea clam extract was conducted on several treatment groups, namely, AOM/DSSinfected mice treated with EtOH extract, ethyl acetate fraction, and N-hexane fraction. The positive control and the negative control of the three treatment groups were done by giving the extract with a dosage of 0.25 mg/bb to mice for 14 days. RESULTS: Based on the result of β-catenin gene amplification using the PCR method and RFLP analysis, the EtOH extract and ethyl acetate fraction groups had the same band, namely, 56 bp and 12 bp, consecutively; meanwhile, the n-hexane fraction was marked by the band formation of 191 bp, 82 bp and 48 bp, 69 bp, 116 bp, 151 bp, 180 bp, and 234 bp. From the types of band and the band size that have been formed, it shows that they have the same formation and similarities in the positive and negative treatment characterized by the amplification that occurred in the band size of 227 bp, and the result of amplification from RFLP analysis showed the same band with the same band size, namely, 89 and 138 bp. CONCLUSION: Based on this, it can be concluded that N-hexane fraction affects β-catenin gene mutation at the first or second base at codon position 32 or 33, and the third base, namely, the bands of 82 and 145 bp that also characterize the effect of N-hexane on β-catenin gene mutation at the second or third base. It is supported by the sequencing method obtaining the homological percentage of 99.13%.