摘要:
通过RT-PCR技术分析红系细胞和非红系细胞RHD的转录情况，同时进一步比较不同D表达个体的RHD的转录关系.采用逆转录PCR（RT-PCR）技术检测HL-60,K562,Jurkat,THP-1,胚肺成纤维细胞系（HECF）,10名不同Rh表型个体的网织红细胞（CcDEe 3名,CCDEe 2名,CCDee 2名,CcDee 2名和CCDuee 1名）以及10名不同Rh表型（2 CCDEe,2 CCDee,2 ccDee,2 CcDee,ccDEe和CcDEe）个体的白细胞的RhD mRNA，然后进行cDNA测序分析.结果表明,HL-60,Jurkat,THP-1,胚肺成纤维细胞系（HECF）以及不同Rh表型个体的外周血有核细胞中除网状红细胞外皆不存在RhD mRNA，K562具有一个正常的RHD基因转录本，而不同表型个体的网织红细胞具有复杂的RhD cDNA形式，有的缺少RHD基因的外显子7,有的缺少外显子7～9或外显子4～9,但这些个体都有一个正常形式的RhD mRNA.由此得出结论,选择性剪切使RHD基因的产生多种形式的转录子，但这些不同形式的转录子仅来自红系的血细胞如网织红细胞和K562细胞系，白细胞、单核细胞、T淋巴细胞以及胚肺成纤维细胞都不具有RHD的mRNA.
Abstract:
This study aimed at analyzing transcription of RHD gene in erythroid cells and non-erythroid cells,and further comparing transcription of RHD gene in individuals with different expression levels of D phenotype.The technique of reverse transcriptase polymorphism chain reaction(RT-PCR) and cDNA sequencing was used to detect RhD mRNA of HL-60,K562,Jurkat,THP-1,HECF and white blood cells from ten unrelated individuals with different Rh phenotypes（2 CCDEe,2 CCDee,2 ccDee,2 CcDee,ccDEe and CcDEe）and reticulocyte from additional ten unrelated individuals with different Rh phenotypes(3 CcDEe,2 CCDEe,2 CCDee，2 CcDee and 1 CCDuee).Results showed except erythroid cells, RhD mRNA was absent in HL-60,Jurkat,THP-1,HECF and nuclear cells of peripheral blood.K562 had a normal RHD transcript.RhD cDNA of reticulocyte from different Rh phenotypes was complex and some absent exon 7, exon 9,exon 7～9,or exon 4～9,but all with normal RHD transcript.Conclusion Multiple RHD transcript is produced by alternative splicing.All these Rh messenger RNA isoforms existing in erythroid cells,such as reticulocyte and K562,absent in white blood cells.
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