THAP11在造血细胞分化中的作用研究

发布时间：2018-01-13 08:19

本文关键词：THAP11在造血细胞分化中的作用研究　出处：《天津大学》2012年硕士论文　论文类型：学位论文

【摘要】：THAP11是THAP蛋白家族的最新成员，前期研究表明THAP11是调控细胞生长、胚胎形成和ES细胞多能性的一个转录因子，THAP11通过下调c-Myc抑制细胞增殖。c-Myc是调控红系巨核系分化的关键转录因子，并且表达谱数据显示THAP11在造血干细胞、多能祖细胞和人脐带血CD34+CD38-细胞中高表达，提示THAP11可能参与造血细胞分化，但目前对于THAP11在造血分化中的作用尚未报道。本论文对THAP11对造血细胞分化的影响进行了深入研究。首先，，通过检测THAP11在人脐带血CD34+细胞向红系和巨核系分化过程中的表达变化，发现THAP11在向红系分化中表达下调，而在巨核分化中表达上调，提示THAP11参与红系/巨核系分化的调控。接着我们构建了THAP11过表达及RNAi的第三代慢病毒载体，通过病毒包装获得慢病毒颗粒，建立了过表达和敲低THAP11的K562和TF-1细胞稳定株。采用hemin诱导K562细胞向红系分化模型，发现THAP11过表达可抑制红系分化，表现为抑制GPA和HBA的上调，联苯胺染色阳性细胞比例降低，而敲低THAP11则促进红系分化。利用EPO诱导TF-1细胞向红系分化的模型，得到相同的结果。采用PMA诱导K562细胞向巨核系分化，发现THAP11过表达可促进巨核系标志物CD61的表达和多倍体形成，与之相反，敲低THAP11则抑制了巨核分化。初步研究表明THAP11可上调GATA-2和Fli1基因，下调c Myc和c-Myb基因的表达，提示THAP11对造血细胞分化的影响可能是通过调控多种基因表达来进行的。另外，过表达THAP11明显抑制CD34+细胞的增殖能力，而敲低THAP11促进CD34+细胞的生长，提示THAP11可能参与了CD34+细胞的生长与增殖。以上结果表明THAP11是一种新的造血细胞分化调控分子，可抑制红系分化，促进巨核系分化。
[Abstract]:THAP11 is the newest member of THAP protein family. Previous studies have shown that THAP11 is a transcription factor regulating cell growth, embryogenesis and es cell pluripotency. THAP11 inhibits cell proliferation by down-regulating c-Myc. C-Myc is the key transcription factor to regulate the differentiation of erythroid megakaryocytes and the expression profile data show that THAP11 plays an important role in hematopoietic stem cells. The overexpression of CD34 CD38- cells in pluripotent progenitor cells and human umbilical cord blood suggests that THAP11 may be involved in hematopoietic cell differentiation. However, the role of THAP11 in hematopoietic differentiation has not been reported. In this paper, the effect of THAP11 on hematopoietic cell differentiation was studied. By detecting the expression of THAP11 in the differentiation of human umbilical cord blood CD34 cells into erythroid and megakaryocyte, it was found that the expression of THAP11 was down-regulated in the differentiation of human umbilical cord blood CD34 cells into erythroid and megakaryocyte. The upregulation in megakaryocyte differentiation suggested that THAP11 was involved in the regulation of erythroid / megakaryocyte differentiation. Then we constructed the third generation lentivirus vector of THAP11 overexpression and RNAi. Lentivirus particles were obtained by virus packaging and stable K562 and TF-1 cell lines with overexpression and knockdown of THAP11 were established. K562 cells were induced into erythroid differentiation model by hemin. It was found that the overexpression of THAP11 could inhibit the differentiation of erythroid cells, which could inhibit the upregulation of GPA and HBA, and decrease the proportion of benzidine staining positive cells. K562 cells were induced to differentiate into megakaryocytes by PMA, and the same results were obtained by using the model of EPO inducing TF-1 cells to differentiate into erythrocytes. It was found that overexpression of THAP11 could promote the expression of megakaryotic marker CD61 and the formation of polyploid. Knockdown of THAP11 inhibited megakaryocyte differentiation. Preliminary studies showed that THAP11 could up-regulate the expression of GATA-2 and Fli1 genes and down-regulate the expression of c Myc and c-Myb genes. The results suggest that the effect of THAP11 on hematopoietic cell differentiation may be mediated by regulating the expression of multiple genes. In addition, overexpression of THAP11 can significantly inhibit the proliferation of CD34 cells. Knockdown of THAP11 promotes the growth of CD34 cells, suggesting that THAP11 may be involved in the growth and proliferation of CD34 cells. These results suggest that THAP11 is a new hematopoietic cell differentiation regulator, which can inhibit erythroid differentiation and promote megakaryocyte differentiation.
【学位授予单位】：天津大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R329

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