Identification of differentially expressed genes in normal mucosa,adenoma and adenocarcinoma of colo

来源 :World Journal of Gastroenterology | 被引量 : 0次 | 上传用户:qqqqq770627
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Abbreviation CRC colorectal cancer.SSHsuppression subtrsctive hybridization.LD PCR longdistance polymerase chain reaction.HLA human leukocyteantigen.IGRBP insulin-like growth factor binding protein.GN guanylin,EF1 elongation factor 1AIM To construct subtracted cDNA libraries and furtheridentify differentially expressed genes that are related tothe development of colorectal carcinoma(CRC).METHODS Suppression subtractive hybridization(SSH)was done on cDNAs of normal mucosa,adenoma andadenocarcinoma tissues from the same patient.Threesubtracted cDNA libraries were constructed and thenhybridized with forward and backward subtracted probesfor differential screening.Positive clones from eachsubtracted cDNA library were selected for sequencing andBLAST analysis.Finally,virtual Northern Blot confirmedsuch differential expression.RESULTS By this way,there were about 3-4×10~2clones identified in each subtracted cDNA library,inwhich about 85% positive clones were differentiallyscreened.Sequencing and BLAST homology searchrevealed some clones containing sequences of knowngene fragments and several possibly novel genes showingfew or no sequence homologies with any knownsequences in the database.CONCLUSION All results confirmed the effectiveness andsensitivity of SSH.The differentially expressed genesduring the development of CRC can be used to shed lighton the pathogenesis of CRC and be useful genetic markersfor early diagnosis and therapy. Abbreviation CRC colorectal cancer.SSHsuppression subtrsctive hybridization.LD PCR longdistance polymerase chain reaction.HLA human leukocyteantigen.IGRBP insulin-like growth factor binding protein.GN guanylin,EF1 elongation factor 1AIM To construct subtracted cDNA libraries and furtheridentify differentially revealed genes that are related tothe Development of colorectal carcinoma (CRC).METHODS Suppression subtractive hybridization(SSH) was done on cDNAs of normal mucosa, adenoma andadenocarcinoma tissues from the same patient.Three subtracted cDNA libraries were constructed and thenhybridized with forward and late subtracted probes for differential screening.Positive clones from Eachsubtracted cDNA library were selected for sequencing and BLAST analysis.Finally, virtual Northern Blot confirmedsuch differential expression.RESULTS By this way,there were about 3-4×10~2clones identified in each subtracted cDNA library,inwhich about 85% positive clones were differentiallyscreened. Sequencin g and BLAST homology searchrevealed some clones containing sequences of knowngene fragments and several possible novel genes showingfew or no sequence homologies with any knownsequences in the database.CONCLUSION All results confirmed the effectiveness andsensitivity of SSH.The differentially represented genesduring the development of CRC can be used To shed lighton the pathogenesis of CRC and be useful genetic markersfor early diagnosis and therapy.
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