DIFFERENTIATION OF TERATOCARCINOMA CELLS: REGULATION

本计画的目标是（1）了解两个基因的结构 其表达受视黄酸控制，和（2）鉴定 参与胶原IV的顺式作用DNA元件和另一个 分化基因在F9干细胞分化中的作用。 我计划 检查这两个F9分化特异性的限制性图谱， 基因，确定结构区域，更具体地说， 转录帽位点。 不同大小的'5侧翼区的两个 基因将与β-珠蛋白结构基因相连，该基因的启动子和'3 基因的末端被删除了。 通过使用杂交的转染测定 基因-SV 40载体作为供体将在F9干细胞中进行。 视黄 将在转染的细胞中测定酸敏感性DNA元件 当检测到β-珠蛋白基因的较高表达程度时， F9细胞分化期。 这一系列的实验将使 我直接鉴定视黄酸调节序列或 间接地。 维甲酸与控制 在上皮组织和一些其他细胞类型中的分化。 的 维甲酸诱导畸胎瘤分化机制 细胞还不太了解。 这一领域的调查推测， 视黄酸可以以类似于类固醇激素的方式起作用。 然而，在这方面， 实验数据还太初步，不能作出任何确切的结论。 的 以分子cDNA为探针， 进一步鉴定视黄酸调节元件将告诉我们， 维甲酸作用的潜在机制。 此外，本发明还提供了一种方法， 对早期胚胎分化的了解也有助于 对肿瘤发生的理解是由细胞的去分化引起的， 一些案件。

The goals of this project are (1) to understand the structure of two genes of which expression is controlled by retinoic acid and (2) to identify cis-acting DNA elements involved in the collagen IV and another differentiation gene during F9 stem cell differentiation. I plan to examine the restriction maps of these two F9 differentiation specific genes, identify the structural regions and more specifically the '5 end transcription cap sites. Different sizes of '5 flanking regions of two genes will be linked to Beta-globin structure gene of which promotor and '3 end of gene has been deleted. Transfection assays by using hybrid gene-SV40 vector as donor will be performed in F9 stem cells. The retinoic acid sensitive DNA elements will be determined in the transfected cells when higher degree of expression of Beta-globin gene is detected at the differentiated stage of F9 cells. These series of experiments will allow me to identify the retinoic acid regulated sequences directly or indirectly. Retinoic acid has been implicated in the control of differentiation in epithelial tissues and in some other cell types. The mechanism by which retinoic acid induce differentiation of teratocarcinoma cells is not well understood. Investigation in this field speculate that retinoic acid may act in a manner analogous to steroid hormones. However, the experimental data is too preliminary to make any firm conclusions. The studies of differentiation genes by using molecular cDNA as probe and further identifications of retinoic acid regulatory elements will inform us of the underlying mechanism of the action of retinoic acid. In addition, the understanding of early embryonic differentiation may also facilitate the understanding of oncogenesis which results from dedifferentiation in some cases.