Wiles M V
Laboratory of Human Molecular Genetics, Imperial Cancer Research Fund, Lincoln's Inn Fields, London, UK.
Development. 1988 Nov;104(3):403-13. doi: 10.1242/dev.104.3.403.
The study of early human development is of great importance but has been limited by the lack of suitable reagents. Recently, however, the human embryonal carcinoma (EC) cell line NT2D1 has been isolated. This cell line will differentiate upon exposure to retinoic acid (RA). A cDNA library was constructed from poly(A)+ RNA derived from NT2D1 cells treated with 10(-5) M-RA for 7 days (delta NT2D1 cells). By differential cDNA screening, it was found that 1.12% of delta NT2D1 cDNA recombinants screened detected an increase in signal with 32P-cDNAs derived from delta NT2D1 as compared with NT2D1. To compare RA-induced differentiation of mouse and human EC cells, the delta NT2D1 cDNA library was rescreened with 32P-cDNAs derived from the mouse EC cell line F9 and the result compared with 32P-cDNA derived from F9 differentiated to parietalendoderm (F9PE)-like cells and visceral-endoderm (F9VE)-like cells. Approximately 1.2% of the delta NT2D1 cDNA recombinants detected a differential increase in signal following differentiation of mouse EC cells to F9VE and/or F9PE. Of these homologous regulated sequences, 0.3% were common to both mouse and human EC cell RA-induced differentiation. Five different cDNA clones were isolated that detect a marked increase (5- to 75-fold) in mRNA abundance following RA-induced differentiation of NT2D1. Of these five clones, three detect homologous mRNAs which also increase in abundance following differentiation of the mouse EC cell line F9 to PE- and/or VE-like cells; the other two clones do not detect sequences in the mouse mRNAs tested. One clone shows homology to SPARC, a gene known to be regulated during mouse embryonic development. While another clone, SO5A, has a limited range of expression, being detected in F9VE and in a human parietal-endoderm-like cell, but not in F9PE and a human visceral-endoderm-like cell. This work shows that there are both similarities and differences in mouse and human EC cell differentiation, and these cDNA clones provide some of the first reagents for studying the molecular biology of human development.
早期人类发育的研究非常重要，但一直受到缺乏合适试剂的限制。然而，最近已分离出人类胚胎癌（EC）细胞系NT2D1。该细胞系在暴露于视黄酸（RA）时会发生分化。用10^(-5)M RA处理NT2D1细胞7天（δNT2D1细胞）后，从其poly(A)+RNA构建了一个cDNA文库。通过差异cDNA筛选发现，与NT2D1相比，在筛选的δNT2D1 cDNA重组体中，有1.12%检测到来自δNT2D1的32P-cDNA信号增强。为了比较RA诱导的小鼠和人类EC细胞的分化，用来自小鼠EC细胞系F9的32P-cDNA对δNT2D1 cDNA文库进行再筛选，并将结果与来自分化为胚外内胚层样细胞（F9PE）和脏内胚层样细胞（F9VE）的F9的32P-cDNA进行比较。大约1.2%的δNT2D1 cDNA重组体在小鼠EC细胞分化为F9VE和/或F9PE后检测到信号差异增强。在这些同源调控序列中，0.3%在小鼠和人类EC细胞RA诱导的分化中是共同的。分离出五个不同的cDNA克隆，它们在NT2D1经RA诱导分化后检测到mRNA丰度显著增加（5至75倍）。在这五个克隆中，三个检测到同源mRNA，在小鼠EC细胞系F9分化为PE和/或VE样细胞后，其丰度也增加；另外两个克隆在测试的小鼠mRNA中未检测到序列。一个克隆与SPARC具有同源性，SPARC是一个已知在小鼠胚胎发育过程中受调控的基因。而另一个克隆SO5A的表达范围有限，在F9VE和人类胚外内胚层样细胞中可检测到，但在F9PE和人类脏内胚层样细胞中未检测到。这项工作表明，小鼠和人类EC细胞分化既有相似之处也有不同之处，这些cDNA克隆为研究人类发育分子生物学提供了首批试剂中的一些。
