CAJ | 학술논문

目的探讨男性特发性不育患者外周血和睾丸组织中无精子因子(AZF)基因表达的临床意义.方法特发性不育患者62例,其中严重少精子症29例.无精子症33例.抽取患者外周血样本检测,8对引物为sY84和sY86(AZFa区).sY127和sY134(AZFb区),sY254和sY255(AZFc区)及内对照SRY(sY14)和ZFY.PCR检测包括MixA:SRY(sY14)-ZFY-sY84-sY134-sY255和MixB:SRY(sY14)-ZFY-sY86-sY127-sY254;穿刺获得患者睾丸标本,Trizol方法提取总RNA,反转录为cDNA.PCR检测包括SRY(sY14)-DFFRY-RBM-DAZ-β-actin.结果外周血PCR结果显示:62例患者中AZF基因微缺失12例(19.4%),其中无精子症组9例,严重少精子症组3例.睾丸组织RT-PCR结果显示:62例均可见SRY阳性表达;RBM mRNA无表达2例,RBM和DAZ mRNA无表达1例,DAZ mRNA无表达12例,其中3例外周血细胞内DAZ基因正常.结论特发性不育患者睾丸组织存在AZF基因表达缺失,睾丸组织RT-PCR检测有助于确定患者病因,结合外周血PCR检测有助于指导睾丸精子穿刺一胞浆内单精子注射治疗.
목적 탐토남성특발성불육환자외주혈화고환조직중무정자인자(AZF)기인표체적림상의의.방법 특발성불육환자62례,기중엄중소정자증29례.무정자증33례.추취환자외주혈양본검측,8대인물위sY84화sY86(AZFa구).sY127화sY134(AZFb구),sY254화sY255(AZFc구)급내대조SRY(sY14)화ZFY.PCR검측포괄MixA:SRY(sY14)-ZFY-sY84-sY134-sY255화MixB:SRY(sY14)-ZFY-sY86-sY127-sY254;천자획득환자고환표본,Trizol방법제취총RNA,반전록위cDNA.PCR검측포괄SRY(sY14)-DFFRY-RBM-DAZ-β-actin.결과 외주혈PCR결과현시:62례환자중AZF기인미결실12례(19.4%),기중무정자증조9례,엄중소정자증조3례.고환조직RT-PCR결과현시:62례균가견SRY양성표체;RBM mRNA무표체2례,RBM화DAZ mRNA무표체1례,DAZ mRNA무표체12례,기중3예외주혈세포내DAZ기인정상.결론 특발성불육환자고환조직존재AZF기인표체결실,고환조직RT-PCR검측유조우학정환자병인,결합외주혈PCR검측유조우지도고환정자천자일포장내단정자주사치료.
Objective To investigate the difference on AZF gene expression between peripheral leukocytes and testicular cells of idiopathic infertility males.Methods Sixty-two idiopathic infertility patients were enrolled.The infertility group included 33 males with azoospermia and 29 with severe oligozoospermia.Genomic DNA was prepared from peripheral blood samples of the 62 patients.Each patient was tested for STS of AZF region located at long arm of Y-chromosome,including sY84 and sY86 for AZFa,sY127 and sY134 for AZFb,sY254 and sY255 for AZFc.The two multiplex PCR consisted of:MixA:SRY(sY14)-ZFY-sY84-sY134-sY255 and MixB:SRY(sY14)-ZFY-sY86-sY127-sY254.The testicular material of infertile men was obtained by bilateral fine needle aspiration cytology.Each patient was tested for STS of AZF region located at long arm of Y-chromosome,including DFFRY for AZFa,RBM for AZFb,sY254(DAZ)for AZFc.The PCR consisted of:SRY(sYl4)-DFFRY-RBM-DAZ(sY254)-β-actin.Results In 62 cases,12 deletions(19.4%)wet found.Nine patients(9/33,27.3%)RT-PCR analysis from testicular cells gave normal amplifications for SRY and DFFRY mRNA in 62 idiopathic patients.Two patients were negative for RBM expression.No RBM and DAZ were detected in 1 case.Twelve patients had no expression in the AZFc region involving the DAZ gene.Of the 12 cases,3 patients with normal PCR analysis of DAZ gene on genomic DNA.Conclusions AZF microdeletion detection is possible in testicular material.Hence RT-PCR analysis can also be helpful to define etiology of idiopathic infertility.