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Folia Medica Indonesiana

ISSN 0303-7932

Vol. 46 / No. 3 / Published : 2010-07

TOC : 7, and page :200 - 205

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Original Article :

Cloning of melanoma antigen-1 (mage-1) gene from fine needle aspiration biopsy of hepatic tissue of hepatocelluler carcinoma patients

Author :

  1. Gondo Mastutik*1
  2. Reny I’tishom*2
  3. JH Lunardi*3
  4. Ni Nyoman Tri Puspaningsih*4
  5. Soetjipto*5
  1. Department of Anatomic Pathology, School of Medicine, Airlangga University
  2. Department of Medical Biology, School of Medicine, Airlangga University
  3. Department of Anatomic Pathology, School of Medicine, Airlangga University
  4. Department of Chemistry, Faculty of Science and Techonology, Airlangga University
  5. Department of Biochemistry, School of Medicine, Airlangga University

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

Keyword :

fine needle aspiration biopsy, hepatocellular carcinoma, melanoma antigen-1, cloning,


References :

  1. Kumar V, Abbas AK, Fausto N, (2005). Robbins and Cotran's Pathologic Basic of Disease. Philadelphia : Elsevier Saunders
  2. De Smet C, Lurquin C, Lethe B, Martelange VR, Boon T, (1999). DNA methylation is the primary silencing mechanism for a set of germ line- and tumor-specific genes with a Cpg-rich promoter. - : Molecular and Cellular Biology
  3. De Smet C, De Backer O, Faraoni I, Lurquin C, Brasseur F, Boon T, (1996). The Activation of human gene MAGE-1 in tumor cells is correlated with genome-wide demethylation. - : Proc Natl Acad Sci


   


Archive Article

Cover Media Content

Volume : 46 / No. : 3 / Pub. : 2010-07
  1. Socioeconomic and genetic factors influencing the strength, weight, length and width measurements of children
  2. Glomerular filtration rate in liver cirrhosis
  3. Antimicrobial activity of mahkota dewa [phaleria macrocarpa (scheff). boerl.] leaf extract against pseudomonas aeruginosa by agar dilution and scanning electron microscopy
  4. Mapping pattern of core dna loci in forensic identification sample examination using polymerase chain reaction (pcr) in loci csf1po, tho1 and tpox
  5. Spatial analysis of dengue fever regarding its determinant factors in bandar lampung
  6. Inhibition of the mitogen activated protein kinase (mapk) in the inflammatory pain like state using sb 203580 and pd 98059 in mice
  7. Cloning of melanoma antigen-1 (mage-1) gene from fine needle aspiration biopsy of hepatic tissue of hepatocelluler carcinoma patients
  8. Detection of urinary incontinence affecting post-menopausal age using ii q-7 and udi-6 questionnaire at immanuel teaching hospital bandung
  9. Electron microscopic scanning profile of balb/c mice intestinal villi after per oral lps (lipopolysaccharide) during probiotic induction
  10. Extensive intensity exercise most effectively increases anaerobic threshold
  11. Pegagan (centella asiatica) extract increases vaginal wall thickness in menopausal rats
  12. Review article: interval exercise with 1:1 work/rest ratio decreases the risk factors of type-2 diabetes mellitus and coronary heart disease
  13. Expression of topoisomerase iiα and cyclin d1 proteins in various degrees of breast ductal carcinoma