Copy Number Status of TERT Gene in a Myeloid Leukaemia Cell Line, K562: Application of Interphase Fluorescence in situ Hybridization (FISH) Analysis

Ahmad Syibli, Othman and Nor Atifah, Mohd Adam and Ravindran, Ankathil and Rosline, Hassan (2011) Copy Number Status of TERT Gene in a Myeloid Leukaemia Cell Line, K562: Application of Interphase Fluorescence in situ Hybridization (FISH) Analysis. Malaysian journal of biochemistry and molecular biology, 19 (1). pp. 12-15. ISSN 1511-2616

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Abstract

Telomeraseisribonucleoproteinenzymecomplexconsistingoftwomaincorecomponents;reversetranscriptase(TERT),andRNAtemplatefortelomericDNAsynthesis(TERC).TERTgeneactsascatalyticcomponentinthisenzymeandislocatedonchromosome5p15.IthasbeenreportedthatthereisanamplificationorincreaseofTERTgenecopynumberinhumancancercelllinesandprimarytumoursampleswhichmayassociatewithincreasedtelomeraseactivity.Inourstudy,dual-colorassayprobewasusedinfluorescencein situhybridization(FISH)analysisthatcoveredthegenomicregionencodingTERT(Redsignal)togetherwithaspecificsequenceofEGR1(Greensignal)at5q31tofacilitatechromosomeidentification.Innormalcells,theFISHsignalpatternwasobservedastwogreensignalsandtworedsignals(2G2R).AmplificationoftheTERTgenehasbeendetectedinmorethan90%ofthecellsincelllineintheleukaemia.Besidesthat,aneusomyofchromosome5wasalsodetectedindicatingthepresenceofthreecopiesofchromosome5with3G3Rsignalpattern.Thus,theapplicationofinterphaseFISHtechniquefordetectionofTERTgeneamplificationhasbeensuccessfullyoptimized

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