促红细胞生成素的抗炎症作用研究进展

促红细胞生成素是一种造血因子,临床中广泛应用于治疗各种原因造成的贫血。近年的研究发现,EPO具有多种非造血生物作用．其中对各组织以及全身炎症反应的保护作用已成为目前的研究热点之一。但目前EPO抗炎症作用相关的机制尚不明确。本文对EPO的抗炎症作用及其可能机制作一综述,并对EPO在临床中的应用前景进行了展望。     

人促红细胞生成素的表达调控

人促红细胞生成素是控制红细胞生长发育的一种重要的细胞因子,在人体内具发育阶段异性、分泌的诱导性等特点,是研究真核基因表达的理想模型。本文从细胞水平的调控、基因结构以及基因３′和５′侧翼序列的调控元件对人促红细胞生成素表达的影响作用进行了讨论。     

红细胞生成素受体功能域的研究及其意义

红细胞生成素受体是细胞因子受体超家族成员之一,它在红细胞生成素诱导的红系祖细胞的存活,增殖,分化的过程中起着重要的调节作用,本文概述了红细胞生成素受体３个功能域的结构,主要功能及其在医药研究中的重要意义。     

母乳中的促红细胞生成素的生理功能

母乳中存在促红细胞生成素.该文介绍母乳中促红细胞生成素的含量以及来源,并着重探讨促红细胞生成素对幼仔的生理作用.     

促红细胞生成素及其基因治疗的研究进展

在简要回顾ＥＰＯ研究进展的基础上，较系统地介绍了ＥＰＯ在贫血基因治疗研究中的进展，最后，对该研究领域存在的问题及前景进行了总结和展望。     

人促红细胞生成素基因的合成及在大肠杆菌中的表达

人促红细胞生成素(简称hEPO)是一种能够促进祖红细胞分化发育并进而促进红细胞生成的细胞生长因子,它在临床上可用以治疗某些贫血病人特别是由于慢性肾衰所引起的肾性贫血^〔1－3〕．hEPO是一种由166个氨基酸组成的糖基化蛋白质．它的前体还带有27个氨基酸的信号肽^〔4－6〕。有两对由Cys7-Cys161和Cys29－Cys33组成的二硫键,其糖基化位点为Ash--24。Ash一38．Asn一83和Ser一126^〔7－8〕。由于天然来源hEPO的量极少．因此通过基因工程的手段是当前获得大量hEPO的较好的途径^〔9－10〕。我们在前文中曾经报道用单链方法合成天花粉胰蛋白酶抑制荆基因〔11〕以及用单链和PCR相结合来合成绿豆胰蛋白酶抑制荆基因的方法〔12〕．我们在合成hEPO基固(包括带有信号肽的hEPO基因)时也采用了这两种方法：合成的hEPO基因在大脑杆菌、CHO细胞和昆虫细胞中均得到表达。本文报道hEPO基因的合成及在大肠杆菌中的表达结果。     

人促红细胞生成素基因在番茄中的表达

通过子叶与农杆菌（ＡｇｒｏｂａｃｔｅｒｉｕｍｔｕｍｅｆａｃｉｅｎｓＡＧＬ１）共培养,将表达载体ｐＨＬＰＥ中的人促红细胞生成素（ｈＥＰＯ,一种人来源的典型糖蛋白）基因（ｅｐｏ）导入番茄,然后用卡那霉素进行筛选,获得了抗性植株。经点杂交和Ｓｏｕｔｈｅｒｎ印迹分析,证明部分抗性植株中整合了ｅｐｏ基因。通过对转基因番茄植株叶片的粗提蛋白进行ＥＰＯＥＬＩＳＡ检测,结果表明,ｈＥＰＯ在番茄中的表达量为约４００ｐｇ／ｇ叶片。经用转基因植株叶片粗提蛋白饲喂依赖于ＥＰＯ的ＴＦ１细胞,表明番茄ＥＰＯ具有体外（ｉｎｖｉｔｒｏ）生物活性,这暗示用植物生产的ＥＰＯ可作为体外药物加以应用。     

人促红细胞生成素在转基因小鼠乳汁中表达

将山羊的β乳球蛋白启动子连接人促红细胞生成素基因,构建转基因表达载体。通过显微注射的方法转基因小鼠。经PCR斑点杂交和Southern杂交检测验证,获得4只转基因阳性小鼠,其中3只母鼠哺乳期收集乳汁,经EpoELISA检测为阳性。对4组转基因阳性小鼠的部分子代母鼠,经PCR和Southern杂交分析,又鉴定出6只获得遗传的阳性G1代母鼠以Dot-ELISA的方法检测,其中两只G1代母鼠乳汁中的Epo含量为0.5μg/mL。实验证实,867bp的β乳球蛋白启动子可以指导人促红细胞生成素在小鼠乳汁中表达。     

红细胞生成素受体研究进展

红细胞生成素受体是促红细胞生成素的作用受体 ,属于细胞因子超家族的成员。近年来 ,由于其克隆表达技术的应用使人们在对造血机制、EPO的信号转导机制 ,及利用其受体筛选 EPO类似物方面的研究有了新的进展。本文综合近年来有关文献对 EPO受体的研究作一概括性介绍。     

The Erythropoietin/Erythropoietin Receptor Signaling Pathway Promotes Growth and Invasion Abilities in Human Renal Carcinoma Cells

Co-expression of erythropoietin (Epo) and erythropoietin receptor (EpoR) has been found in various non-hematopoietic cancers including hereditary and sporadic renal cell carcinomas (RCC), but the Epo/EpoR autocrine and paracrine mechanisms in tumor progression have not yet been identified. In this study, we used RNA interference method to down-regulate EpoR to investigate the function of Epo/EpoR pathway in human RCC cells. Epo and EpoR co-expressed in primary renal cancer cells and 6 human RCC cell lines. EpoR signaling was constitutionally phosphorylated in primary renal cancer cells, 786-0 and Caki-1 cells, and recombinant human Epo (rhEpo) stimulation had no significant effects on further phosphorylation of EpoR pathway, proliferation, and invasiveness of the cells. Down-regulation of EpoR expression in 786-0 cells by lentivirus-introduced siRNA resulted in inhibition of growth and invasiveness in vitro and in vivo, and promotion of cell apoptosis. In addition, rhEpo stimulation slightly antagonized the anti-tumor effect of Sunitinib on 786-0 cells. Sunitinib could induce more apoptotic cells in 786-0 cells with knockdown EpoR expression. Our results suggested that Epo/EpoR pathway was involved in cell growth, invasion, survival, and sensitivity to the multi-kinases inhibitor Sunitinib in RCC cells.     

人红细胞生成素受体的研究进展

人红细胞生成素受体(hEPOR)是位于相对成熟阶段人体红系祖细胞表面的跨膜蛋白,它能专一性结合人红细胞生成素(hEPO),将促进细胞生长、增殖和分化的信号传导到膜内,该过程涉及了hEPOR自身及部分相关蛋白的磷酸化.hEPOR具有一些与其功能相关的保守结构,它的氨基酸序列与鼠EPOR(mEPOP)高度同源.EPOR因为膜外R129C突变或结合特定蛋白质而具有组成型活性.EPOR还与红白血病等多种血液病密切相关.     

Erythropoietin Receptor Expression Is a Potential Prognostic Factor in Human Lung Adenocarcinoma

Recombinant human erythropoietins (rHuEPOs) are used to treat cancer-related anemia. Recent preclinical studies and clinical trials, however, have raised concerns about the potential tumor-promoting effects of these drugs. Because the clinical significance of erythropoietin receptor (EPOR) signaling in human non-small cell lung cancer (NSCLC) also remains controversial, our aim was to study whether EPO treatment modifies tumor growth and if EPOR expression has an impact on the clinical behavior of this malignancy. A total of 43 patients with stage III–IV adenocarcinoma (ADC) and complete clinicopathological data were included. EPOR expression in human ADC samples and cell lines was measured by quantitative real-time polymerase chain reaction. Effects of exogenous rHuEPOα were studied on human lung ADC cell lines in vitro. In vivo growth of human ADC xenografts treated with rHuEPOα with or without chemotherapy was also assessed. In vivo tumor and endothelial cell (EC) proliferation was determined by 5-bromo-2’-deoxy-uridine (BrdU) incorporation and immunofluorescent labeling. Although EPOR mRNA was expressed in all of the three investigated ADC cell lines, rHuEPOα treatment (either alone or in combination with gemcitabine) did not alter ADC cell proliferation in vitro. However, rHuEPOα significantly decreased tumor cell proliferation and growth of human H1975 lung ADC xenografts. At the same time, rHuEPOα treatment of H1975 tumors resulted in accelerated tumor endothelial cell proliferation. Moreover, in patients with advanced stage lung ADC, high intratumoral EPOR mRNA levels were associated with significantly increased overall survival. This study reveals high EPOR level as a potential novel positive prognostic marker in human lung ADC.     

Expression of Asialoglycoprotein Receptor 1 in Human Hepatocellular Carcinoma

Human hepatocellular carcinoma (HCC) is the fifth most common cancer in the world. Currently, surgical resection is the only effective treatment for HCC if the tumor is resectable. Small molecule, biologics and siRNA anti-cancer drugs have been explored for the treatment of HCC. Selective targeting to tumor tissue rather than normal liver in HCC patients is still a challenge. Galactosamine-mediated targeting delivery of anti-cancer drugs in the liver has been tested because its receptor, asialoglycoprotein receptor 1 (ASGPR1), is expressed in the liver and not in other human tissues. We examined ASGPR1 expression levels by immunohistochemistry in HCC with different grades. Guidance for a targeting delivery strategy for anti-cancer drugs to HCC is suggested in this report.     

人促红细胞生成素受体胞外区基因的克隆及其在大肠肝菌中的表达

以人胎肝为材料,通过ＰＴ－ＰＣＲ的方法扩增出人促红细胞生成素受体（ｈＰｅｏＲ）的胞外区基因。将获得的或熟体胞外区基因起始密码子改构后克隆原核表达载体ｐＢＶ２２０中,进行原核温控诱导表达。表达产物经蛋白Ｎ端测序及Ｗｅｓｔｅｒｎ－ｂｌｏｔ实验证实表达产物是ｈＥｐｏＲ胞外区蛋白。利用上罐发酵培养获得的包涵体蛋白经复性纯化后,体外生物学活性检测表明表达产物可特 抑制ＴＦ－１细胞在Ｅｐｏ刺激下的生长,证实了     

HoxA10 Induces Proliferation in Human Prostate Carcinoma PC-3 Cell Line

The objectives of this study were to examine the expression levels of Homeobox A10 (HoxA10) in prostate cancer cells and to study the molecular mechanism of HoxA10-mediated regulation of prostate cancer cell growth and development. We investigated the effect of HoxA10 on cell proliferation by stably overexpressing or silencing HoxA10 in prostate cancer PC-3 cell line using lentiviral vectors. Quantitative real-time PCR and western blotting analysis were used to compare the expressions of HoxA10 in prostate cancer cell lines and normal prostate epithelium. Cancer cell proliferation was examined by MTT assay and colony formation assay. The levels of HoxA10 expression were significantly increased in prostate cancer cell lines and tissues compared to those in normal prostate epithelium. Overexpression of HoxA10 in PC-3 cells induced significant cancer cell proliferation, whereas silencing of HoxA10 expression by RNAi resulted in decreased proliferation rates. HoxA10 was highly expressed in prostate cancer cells and tissues, suggesting its functional involvement in cancer cell proliferation. We successfully overexpressed or silenced HoxA10 in prostate cancer PC-3 cell line and discovered that the levels of HoxA10 directly correlate with cancer cell proliferation. These findings contribute to a better understanding of the regulatory mechanism of HoxA10 in prostate cancer.