CLONING OF MELANOMA ANTIGEN-1 (MAGE-1) GENE FROM FINE NEEDLE ASPIRATION BIOPSY OF HEPATIC TISSUE OF HEPATOCELLULER CARCINOMA PATIENTS

Gondo Mastutik and Reny I'tishom and JH Lunardi and Ni Nyoman Tri Puspaningsih and Soetjipto and Hernomo Ontoseno and Suhartono Taat Putra (2010) CLONING OF MELANOMA ANTIGEN-1 (MAGE-1) GENE FROM FINE NEEDLE ASPIRATION BIOPSY OF HEPATIC TISSUE OF HEPATOCELLULER CARCINOMA PATIENTS. Folia Medica Indonesiana, 46 (3). pp. 200-215. ISSN 0303-7932

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Abstract

Hepatocellular carcinoma (HCC) is one of the most common malignancies in Asia. HCC expresses mRNA of melanoma antigen-1 (MAGE-1) and not expressed in non-tumor hepatic cells. This expression has been taking place at the early malignant transformation and continuously undergoing its progression. MAGE-1 is the important marker for diagnose HCC. MAGE-1 protein could be detected with antibody to MAGE-1 protein which could be obtained by doing clone of MAGE-1. However, the recombinant plasmid of MAGE-1 clone for developing of the diagnosis of HCC hasn’t been obtained. The objective was to clone of coding sequence (CDS) of MAGE-1 from fine needle aspiration biopsy (FNAB) of HCC into vector and to obtain MAGE-1 recombinant plasmid. cDNA of MAGE-1 were extracted then continued by RT PCR and nested PCR. The PCR product was cloned to pET101/D-TOPO and transformed to E. coli Top10. Analyze of recombinant plasmid was undertaken by sequencing and restriction test. The result revealed that CDS of MAGE-1 were isolated and obtained band +1105 bp at first round and +931 bp at second round. The recombinant plasmid pETGM/MAGE1-HCC contained 927 nucleotides that encoded 309 amino acids. Partial sequence of MAGE-1 can be accessed in GeneBank EU161102 and ABW06861. Analyze of CDS MAGE-1 from HCC and GeneBank had 100% homology with M77481 and NM_004988 (both from skin melanoma tissue) and 99% with BC017555 (from skin melanoma tissue) and AY148486 (from HCC). This study obtains recombinant plasmid pETGM/MAGE1-HCC from the FNAB of HCC. It reveals that MAGE-1 recombinant plasmid that can be used for developing the diagnosis of HCC

Item Type: Article
Uncontrolled Keywords: fine needle aspiration biopsy, hepatocellular carcinoma, melanoma antigen-1, cloning
Subjects: R Medicine > R Medicine (General) > R735-854 Medical education. Medical schools. Research
Creators:
CreatorsNIM/NIDN
Gondo Mastutikgondomastutik@gmail.com
Reny I'tishomUNSPECIFIED
JH LunardiUNSPECIFIED
Ni Nyoman Tri PuspaningsihUNSPECIFIED
SoetjiptoUNSPECIFIED
Hernomo OntosenoUNSPECIFIED
Suhartono Taat PutraUNSPECIFIED
Depositing User: Bayu Yulianti
Date Deposited: 21 Aug 2017 20:36
Last Modified: 16 Jul 2019 07:41
URI: http://repository.unair.ac.id/id/eprint/60735
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