Rare and Frequent Promoter Methylation, Respectively, of TSHZ2 and 3 Genes That Are Both Downregulated in Expression in Breast and Prostate Cancers 英文参考文献.docVIP

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Rare and Frequent Promoter Methylation, Respectively, of TSHZ2 and 3 Genes That Are Both Downregulated in Expression in Breast and Prostate Cancers 英文参考文献.doc

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Rare and Frequent Promoter Methylation, Respectively, of TSHZ2 and 3 Genes That Are Both Downregulated in Expression in Breast and Prostate Cancers 英文参考文献

RareandFrequentPromoterMethylation,Respectively, ofTSHZ2and3GenesThatAreBothDownregulatedin ExpressioninBreastandProstateCancers MiyakoYamamoto1,2*,EmiliCid2,SamuelBru2,FumiichiroYamamoto1,2 1Burnham Institute for Medical Research (BIMR), La Jolla, California, United States of America, 2Institute of Predictive and Personalized Medicine of Cancer (IMPPC), Badalona,Barcelona,Spain Abstract Background:NeoplasticcellsharborbothhypomethylatedandhypermethylatedregionsofDNA.Whereashypomethyla- tion is found mainly in repeat sequences, regional hypermethylation has been linked to the transcriptional silencing of certaintumorsuppressorgenes.Weattemptedtosearchforcandidategenesinvolvedinbreast/prostatecarcinogenesis, using the criteria that they should be expressed in primary cultures of normal breast/prostate epithelial cells but are frequentlydownregulatedinbreast/prostatecancercelllinesandthattheirpromotersarehypermethylated. Methodology/Principal Findings: We identified several dozens of candidates among 194 homeobox and related genes usingSystematicMultiplexRT-PCRandamong23,000knowngenesand23,000otherexpressedsequencesinthehuman genomebyDNAmicroarrayhybridization.Anadditionalexamination,byreal-timeqRT-PCRofclinicalspecimensofbreast cancer,furthernarrowedthelistofthecandidates.Amongthem,themostfrequentlydownregulatedgenesintumorswere NP_775756andZNF537,fromthehomeoboxgenesearchandthegenome-widesearch,respectively.Tooursurprise,we laterdiscoveredthatthesegenesbelongtothesamegenefamily,the3-memberTeashirtfamily,bearingthenewnamesof TSHZ2andTSHZ3.Wesubsequentlydeterminedthemethylationstatusoftheirgenepromoters.TheTSHZ3genepromoter wasfoundtobemethylatedinallthebreast/prostatecancercelllinesandsomeofthebreastcancerclinicalspecimens analyzed.TheTSHZ2genepromoter,ontheotherhand,wasunmethylatedexceptfortheMDA-MB-231breastcancercell line.TheTSHZ1genewasalwaysexpressed,anditspromoterwasunmethylatedinallcases. Conclusions/Significance: TSHZ2 and TSHZ3 genes turned out to be the m