正常与异常核型人类胚胎干细胞体外分化不同时期基因表达的异同  

Different gene expression at different times after spontaneous differentiation in hESC lines with normal and abnormal karyotype

作　　者：刘维强[1,2] 孙筱放[1,2] 银益飞[1] 骆玉梅[1,2] 蒋永华[1,2] 李少英[1,2] 郑育红[1,2] 孔舒[1,2] 

机构地区：[1]广州医学院第三附属医院,广东广州510150 [2]广州市生殖与遗传重点实验室,广东广州510150

出　　处：《现代生物医学进展》2009年第18期3422-3426,F0003,共6页Progress in Modern Biomedicine

基　　金：国家自然基金项目(30871378)

摘　　要：目的:研究正常核型和异常核型人胚胎干细胞(hESC)基因的表达异同。方法:实时荧光相对定量PCR检测两株正常核型(46,XX)及一株平衡易位13三体核型和一株三倍体核型hESC在体外自体分化不同时期X连锁基因PGK1、抑癌基因RBBP7及癌症基因GPC4的表达情况,并比较分化后不同时期、不同核型对父系印迹基因H19、IGF2R,母系印迹基因SNRPN及多能性调控基因OCT4、NANOG的影响。结果:随着分化时间的增加:①正常和异常核型hESC的PGK1均上调表达;②异常核型hESC抑癌基因RBBP7及癌症基因GPC4相对正常核型hESC呈现明显上调表达;③正常和异常核型hESC中印迹基因表达基本一致:H19、IGF2R上调而SNRPN表达变化不明显或下调;④多能性调控基因OCT4、NANOG在正常核型hESC中较在异常核型hESC中表达明显下降。结论:X连锁基因PGK1、印迹基因在hESC的发育过程中能维持正常调节而不受核型的影响。异常核型hESC抑癌基因、癌症基因在发育过程中的表达上调表明此种细胞具有更危险的发育前景,同时多能性基因在分化后仍能检出表明此种细胞分化能力较正常细胞弱。Objective： To investigate different genes expression in human embryonic stem cell （hESC） lines with normal and ab- normal karyotype after differentiation. Methods： Relative quantitative real-time PCR was used to detect the temporal expression of gene PGK1, X chromosome tumor suppressor gene RBBP7, oncogene GPC4 as well as paternally imprinted gene H19, IGF2R and maternally imprinted gene SNRPN at different times after spontaneous differentiation in two normal and two abnormal karyotype hESC lines. The expression of OCT4, NANOG was also investigated. Results： ①PGK1 showed up-regulated expression in all hESCs. ② Compared to the normal hESCs, RBBP7 and GPC4 showed significantly up-regulated expression in the abnormal hESCs.③Similar imprinted gene expression patterns were found in both normal and abnormal hESCs. H19, IGF2R showed up-regulated expression following the time after differentiation while SNRPN showed down-regulated. ④The expression of OCT4, NANOG was significantly decreased in the normal hESCs after differentiation compared to the abnormal hESCs. Conclusion： The expression patterns of PGK1 and imprinted genes in normal and abnormal hESCs were similar, which indicates that an appropriate order of those genes in hESCs. The stronger self-renewal and weaker differentiation ability and the up-regulated expression levels in oncogene and tumor suppressor gene in abnormal hESCs may be a great hurdle to the development of those cells.

关 键 词：胚胎干细胞 核型 印迹基因 抑癌基因 癌症基因 

分 类 号：Q344[生物学—遗传学]