【摘 要】
：
目的：在验证不同浓度叶酸培养影响人正常肝细胞系增殖、周期和迁移能力的基础上,应用RNA-Seq技术筛选差异表达基因,为进一步探讨叶酸缺乏与基因差异表达以及正常肝细胞恶性转化之间的相关性打下基础.方法：不同浓度叶酸培养(无叶酸组：0mg/L,正常叶酸组：40mg/L)的人正常肝细胞系QSG-7701 6个月,应用CCK-8实验检测细胞增殖、流式细胞技术检测细胞周期、Transwell小室检测细胞迁移
【机 构】
：
第二军医大学基础部医学遗传学教研室 上海200433
论文部分内容阅读
目的：在验证不同浓度叶酸培养影响人正常肝细胞系增殖、周期和迁移能力的基础上,应用RNA-Seq技术筛选差异表达基因,为进一步探讨叶酸缺乏与基因差异表达以及正常肝细胞恶性转化之间的相关性打下基础.方法：不同浓度叶酸培养(无叶酸组：0mg/L,正常叶酸组：40mg/L)的人正常肝细胞系QSG-7701 6个月,应用CCK-8实验检测细胞增殖、流式细胞技术检测细胞周期、Transwell小室检测细胞迁移；利用RNA-Seq技术对两组细胞中的mRNA进行检测,筛选差异mRNA；通过用实时定量PCR对RNA-Seq的结果进行验证,并结合统计学和生物信息学方法分析差异mRNA.结果：无叶酸培养显著提高QSG-7701细胞的增殖能力,促进细胞由静止期进入分裂期,促进细胞的迁移能力；RNASeq检测得到含有16774条差异mRNA的数据集,初步分析显示其中391条差异mRNA可能与不同浓度叶酸培养相关,其中上调的1 15条,下调的276条；随机对其中1 1条进行实时定量PCR验证,72.73％(8)的结果与RNA-Seq一致；分析显示,差异mRNA相对应的基因与细胞周期等生理过程以及多种肿瘤的发生、发展密切相关.结论：最终筛选出参与叶酸调控正常肝细胞恶性转化相关的基因,RNA-Seq技术能有效的用于差异基因的筛选.
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