秦巴山区精神发育迟滞患者家系人群B淋巴细胞永生细胞库的建立及影响因素初探

发布时间：2018-04-04 12:44

本文选题：精神发育迟滞　切入点：EB病毒　出处：《西北大学》2013年硕士论文

【摘要】：精神发育迟滞(Mental retardation, MR)是一类神经与精神疾病,在人群中发病率为1-3%,带给患者、家庭及社会沉重的经济与精神负担。发病因素涉及环境和遗传因素,对其致病机理研究方兴未艾。用EB病毒体外转化人外周血B淋巴细胞,建立永生细胞库,可以源源不断的为各种复杂的遗传性疾病致病机理研究提供大量材料(细胞、DNA和蛋白质),可扩大和保存一些珍贵的遗传资源,为进一步研究致病机理和干预奠定基础。本研究在知情同意的原则下,收集了141例精神发育迟滞患者及家系成员外周血样品,利用EB病毒体外转化技术,将MR家系人群外周血B淋巴细胞转化为类淋巴母细胞系,建立永生细胞库。建系成功后,采用遗传学方法对其进行鉴定。结果发现,淋巴母细胞系能够表达EB病毒基因,可连续传代,扩大培养,并且保持了成熟B淋巴细胞的生物学特性。对建库成功后的细胞株进行了复苏和传代培养,结果发现在传到第17代时,细胞生长速度开始变慢,经半定量RT-PCR检测,我们发现cyclin D1、cdc2、p53和c-myc基因的表达出现异常,推测其可能是导致细胞传代过程中增殖速度发生变化的主要原因。
[Abstract]:Mental retardation (MRM) is a kind of neurologic and mental disease, which causes a heavy economic and mental burden to patients, families and society.The pathogenic factors are related to environmental and genetic factors, and the research on the pathogenesis of them is in the ascendant.Human peripheral blood B lymphocytes were transformed with Epstein-Barr virus (EBV) in vitro, and the immortalized cell bank was established.It can provide a large number of materials (cell DNA and protein) for the study of the pathogenesis of various complex genetic diseases. It can expand and preserve some precious genetic resources and lay a foundation for further research on the pathogenicity mechanism and intervention.Based on the principle of informed consent, peripheral blood samples of 141 patients with mental retardation and their family members were collected and transformed into lymphoblastoid cell lines (LMCs) by Epstein-Barr virus (EBV) transformation in vitro.Establish an immortalized cell bank.After the successful establishment of the system, genetic methods were used to identify it.The results showed that the lymphoblastoid cell line could express EB virus gene, be subcultured continuously, expand culture, and maintain the biological characteristics of mature B lymphocytes.The successful cell lines were resuscitated and subcultured. The results showed that the cell growth rate began to slow down at the 17th generation. By semi-quantitative RT-PCR, we found the abnormal expression of p53 and c-myc genes in the cyclin D1 cdc2 gene.It is speculated that it may be the main reason for the change of proliferation rate in the course of cell passage.
【学位授予单位】：西北大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R749.93;R394

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