鸟氨酸脱羧酶抗酶I与多胺代谢

鸟氨酸脱羧酶抗酶I（ornithine decarboxylase antizyme 1,OAZ1）是调节细胞内多胺含量的重要因子,参与细胞生长与分化、胚胎发育、基因表达调控等重要生理过程,也是抗肿瘤治疗的潜在分子靶点。简要综述鸟氨酸脱羧酶抗酶I在结构与功能,及其调控多胺代谢机制方面的研究进展。     

鸟氨酸脱羧酶抗酶抑制因子-1与细胞增殖

多胺(腐胺、精脒、精胺)参与细胞增殖、分化和凋亡等重要生命过程,细胞内多胺代谢紊乱也与包括肿瘤在内的多种疾病的发生发展密切相关。鸟氨酸脱羧酶抗酶抑制因子-1(antizyme inhibitor-1,AZIN1)是重要的多胺代谢调节蛋白,它通过多种途径调控细胞的生长。AZIN1能与鸟氨酸脱羧酶抗酶(antizyme,AZ)相互作用,解除后者对多胺合成限速酶鸟氨酸脱羧酶(ornithine decarboxylase,ODC)的抑制,由此上调细胞内多胺的含量。AZIN1还能通过调节细胞周期蛋白cyclin D1的降解速度和干扰细胞中心体复制而影响细胞增殖。AZIN1基因转录后修饰和某些特定mi RNA也对AZIN1的细胞增殖调节功能有重要影响。现对该研究领域的研究进展做简要综述。     

多胺代谢与调节

多胺为阳离子脂肪族胺,在细胞生长、增殖和分化过程中具有重要的调控作用,主要参与DNA、RNA和蛋白质的合成及稳定性的调节以及细胞膜功能、酶功能及环磷酸核苷代谢等过程的调节。在另一方面,多胺代谢中的一些酶含量低,半衰期短,易于被诱导,因此又可受到体内许多因素如激素、体液因子的调节,使多胺维持在一定水平并保持多胺各成分之间的正常比例,保证细胞在正常水平增殖分化。本文着重介绍了多胺的合成代谢、相互转化及分解代谢的途径和参与这些代谢的酶的一些特性及其调节这些酶的各种因素,并探讨了多胺代谢在基础研究及临床应用上的意义。     

多胺在癌症治疗中的作用及机制

多胺是真核细胞生长及发育的必需物质,多胺代谢功能的紊乱与癌症发生密切相关。研究表明,抑制多胺生物合成途径的限速酶鸟氨酸脱羧酶和S-腺苷甲硫氨酸脱羧酶能有效缓解癌症的发展。此外,利用多胺跨膜转运系统的特异性,可将多胺类似物和缀合物转运至细胞内,通过降低细胞内多胺水平,调节组蛋白乙酰化和甲基化水平,促进肿瘤细胞凋亡等途径发挥其抗癌治疗的作用。综述了通过抑制多胺合成酶以及利用多胺类似物和缀合物治疗癌症的研究进展,以期为今后利用多胺代谢途径靶向治疗癌症的研究提供参考。     

《多胺生物学与化学》

《多胺生物学与化学》(The biology and chemistry of polyamines)由Sara H. Goldemberg和Israel D. Algranati编著,1990年IRL Press出版,244页。该书有下列部份:大分子合成和代谢调节中的多胺;酶和多胺代谢调节;微生物和病毒中的多胺;寄生物和植物中的多胺;细胞生长和分化中的多胺;多胺衍生物合成设计与代谢     

多胺在肿瘤研究中的意义

多胺（Ｐｏｌｙａｍｉｎｅ）是一类含二个或二个以上氨基的脂肪族化合物,它包括腐胺（Ｐｕｔｒｅｓｃｉｎｅ,Ｐｕｔ）、精脒（亚精胺）（Ｓｐｅｒｍｉｄｉｎｅ,Ｓｐｄ）、精胺（ｓｐｅｒｍｉｎｅｓｐｍ）。多胺来源于Ｌ－鸟氨酸;鸟氨酸脱羧酶（ＯｒｎｉｔｈｉｎｅＤｅｃａｒｂｏｘｙｌａｓｅ,ＯＤＣ）是多胺合成的限速酶。多胺与细胞的分裂分化、核酸代谢、蛋白质生物合成等有密切关系。细胞外液中浓度升高的多胺主要来源于肿瘤细胞。现已发现神经系统肿瘤、白血病、淋巴瘤、恶性黑色素瘤、肺癌、乳腺癌、消化系统肿瘤、泌尿生殖系统肿瘤病人体液中的多胺含量有不同程度的升高,作为恶性肿瘤辅助诊断、疗效及预后的制定,是一项较好的（临床诊断肿瘤）指标。多胺的分析方法、多胺合成抑制剂、鸟氨酸脱羧酶的调控机制及多胺与癌基因之间关系是值得进一步研究的新颖内容。     

胍丁胺酶与相关疾病关系的研究进展

胍丁胺酶(agmatinase,AGM)属于精氨酸酶家族中的一员,它是一类Mn~(2+)依赖的脲水解酶。其主要作用是调节底物胍丁胺的水平,并参与多胺旁路合成途径调节多胺含量。胍丁胺酶在人体多种组织中差异表达,研究表明其表达异常与乙肝、抑郁症、白血病及肿瘤等多种疾病的发生发展有着紧密联系。因此,进一步阐明胍丁胺酶与相关疾病之间的分子机制意义重大。本文主要介绍了胍丁胺酶与相关疾病的关系,旨在为疾病防治提供新的思路。     

细胞内胆固醇水平调节研究进展

细胞内胆固醇水平动态平衡是细胞发挥生理功能的重要保障.破坏细胞内胆固醇水平动态平衡不仅增加心血管系统疾病患病风险,而且与许多代谢性疾病相关.细胞内胆固醇水平主要受胆固醇生物合成、摄取、流出和酯化的调节.3-羟基-3-甲基-戊二酰基辅酶A还原酶、角鲨烯单加氧酶和固醇调节元件结合蛋白2是胆固醇合成关键因子.尼曼-匹克C1型...     

多胺对神经退行性疾病的改善作用

多胺(polyamines)普遍存在于各种生物体中,参与多种生物学功能,包括细胞生长和增殖、蛋白质和核酸的合成、免疫细胞的分化、炎症反应的调节以及肠道功能的维持等。多胺主要包括腐胺(putrescine)、亚精胺(spermidine)、精胺(spermine)以及胍丁胺(agmatine)。随着衰老进程的发展,细胞内的多胺水平会逐渐随之降低。在衰老相关的神经退行性疾病的发生和发展过程中,多胺具有积极的作用。因此,本文就多胺对神经退行性疾病的改善作用作一综述,期望可以对未来神经退行性疾病的治疗和缓解提供参考。     

多胺对菊花激素含量与花芽分化的影响

以切花菊（Dendranthema morifolium）品种‘神马’为试材,外源喷施0．1mmol·L-1的亚精胺（Spd）与多胺合成抑制剂D-精氨酸（D．Arg）,转入昼10h／夜14h的短日条件下进行开花诱导,测定不同花芽分化时期顶芽内源多胺[腐胺（Put）、亚精胺（Spd）、精胺（spm）]和几种激素[生长素（IAA）、玉米素核苷（ZR）、异戊烯基腺苷（iPA）、赤霉素（GA）]含量的动态变化,分析多胺对激素和花芽分化的作用关系。结果表明,外源多胺和多胺合成抑制剂能够显著影响顶芽内源多胺（Put、Spd、Spm）和激素（IAA、ZR、iPA、GA）的含量,顶芽内高水平多胺有利于菊花花芽分化的启动和保持;外源多胺及多胺合成抑制剂可能通过影响内源多胺含量从而影响内源激素或者直接影响内源激素和内源多胺,进而调控花芽分化：内源Put与IAA关系密切,高水平的内源Put不利于IAA的积累;ZR和iPA含量与内源多胺总量的变化趋势一致;外源多胺及多胺合成抑制剂对GA的影响主要在花序分化期和小花分化期,且高水平的内源Spd和Put不利于GA的积累。     

Evidence of a role for antizyme and antizyme inhibitor as regulators of human cancer

Antizyme and its endogenous antizyme inhibitor have recently emerged as prominent regulators of cell growth, transformation, centrosome duplication, and tumorigenesis. Antizyme was originally isolated as a negative modulator of the enzyme ornithine decarboxylase (ODC), an essential component of the polyamine biosynthetic pathway. Antizyme binds ODC and facilitates proteasomal ODC degradation. Antizyme also facilitates degradation of a set of cell cycle regulatory proteins, including cyclin D1, Smad1, and Aurora A kinase, as well as Mps1, a protein that regulates centrosome duplication. Antizyme has been reported to function as a tumor suppressor and to negatively regulate tumor cell proliferation and transformation. Antizyme inhibitor binds to antizyme and suppresses its known functions, leading to increased polyamine synthesis, increased cell proliferation, and increased transformation and tumorigenesis. Gene array studies show antizyme inhibitor to be amplified in cancers of the ovary, breast, and prostate. In this review, we summarize the current literature on the role of antizyme and antizyme inhibitor in cancer, discuss how the ratio of antizyme to antizyme inhibitor can influence tumor growth, and suggest strategies to target this axis for tumor prevention and treatment.     

Antizyme targets cyclin D1 for degradation. A novel mechanism for cell growth repression

Overproduction of the ornithine decarboxylase (ODC) regulatory protein ODC-antizyme has been shown to correlate with cell growth inhibition in a variety of different cell types. Although the exact mechanism of this growth inhibition is not known, it has been attributed to the effect of antizyme on polyamine metabolism. Antizyme binds directly to ODC, targeting ODC for ubiquitin-independent degradation by the 26 S proteasome. We now show that antizyme induction also leads to degradation of the cell cycle regulatory protein cyclin D1. We demonstrate that antizyme is capable of specific, noncovalent association with cyclin D1 and that this interaction accelerates cyclin D1 degradation in vitro in the presence of only antizyme, cyclin D1, purified 26 S proteasomes, and ATP. In vivo, antizyme up-regulation induced either by the polyamine spermine or by antizyme overexpression causes reduction of intracellular cyclin D1 levels. The antizyme-mediated pathway for cyclin D1 degradation is independent of the previously characterized phosphorylation- and ubiquitination-dependent pathway, because antizyme up-regulation induces the degradation of a cyclin D1 mutant (T286A) that abrogates its ubiquitination. We propose that antizyme-mediated degradation of cyclin D1 by the proteasome may provide an explanation for the repression of cell growth following antizyme up-regulation.     

Stable siRNA-mediated silencing of antizyme inhibitor: regulation of ornithine decarboxylase activity

Ornithine decarboxylase (ODC) is the rate-limiting enzyme involved in the biosynthesis of polyamines essential for cell growth and differentiation. Aberrant upregulation of ODC, however, is widely believed to be a contributing factor in tumorigenesis. Antizyme is a major regulator of ODC, inhibiting ODC activity through the formation of complexes and facilitating degradation of ODC by the 26S proteasome. Moreover, the antizyme inhibitor (AZI) serves as another factor in regulating ODC, by binding to antizyme and releasing ODC from ODC-antizyme complexes. In our previous report, we observed elevated AZI expression in tumor specimens. Therefore, to evaluate the role of AZI in regulating ODC activity in tumors, we successfully down-regulated AZI expression using RNA interference technology in A549 lung cancer cells expressing high levels of AZI. Two AZI siRNAs, which were capable to generate a hairpin dsRNA loop targeting AZI, could successively decrease the expression of AZI. Using biological assays, antizyme activity increased in AZI-siRNA-transfected cells, and ODC levels and activity were reduced as well. Moreover, silencing AZI expression decreased intracellular polyamine levels, reduced cell proliferation, and prolonged population doubling time. Our results directly demonstrate that downregulation of AZI regulates ODC activity, intracellular polyamine levels, and cell growth through regulating antizyme activity. This study also suggests that highly expressed AZI may be partly responsible for increased ODC activity and cellular transformation.     

Solution structure of a conserved domain of antizyme: a protein regulator of polyamines

Antizyme and its isoforms are members of an unusual yet broadly conserved family of proteins, with roles in regulating polyamine levels within cells. Antizyme has the ability to bind and inhibit the enzyme ornithine decarboxylase (ODC), targeting it for degradation at the proteasome; antizyme is also known to affect the transport of polyamines and interact with the antizyme inhibitor protein (AZI), as well as the cell-cycle protein cyclin D1. In the present work, NMR methods were used to determine the solution structure of a stable, folded domain of mammalian antizyme isoform-1 (AZ-1), consisting of amino acid residues 87-227. The protein was found to contain eight beta strands and two alpha helices, with the strands forming a mixed parallel and antiparallel beta sheet. At the level of primary sequence, antizyme is not similar to any protein of known structure, and results show that antizyme exhibits a novel arrangement of its strands and helices. Interestingly, however, the fold of antizyme is similar to that found in a family of acetyl transferases, as well as translation initiation factor IF3, despite a lack of functional relatedness between these proteins. Structural results, combined with amino acid sequence comparisons, were used to identify conserved features among the various homologues of antizyme and their isoforms. Conserved surface residues, including a cluster of acidic amino acids, were found to be located on a single face of antizyme, suggesting this surface is a possible site of interaction with target proteins such as ODC. This structural model provides an essential framework for an improved future understanding of how the different parts of antizyme play their roles in polyamine regulation.     

Ornithine decarboxylase antizyme in kidneys of male and female mice.

Antizyme, a protein inhibitor of ornithine decarboxylase (ODC), was shown to be induced in mouse kidney by repeated injection of putrescine. Antizyme was also present as a complex with ODC in the kidney of untreated mouse. The amount of the renal ODC-antizyme complex was 3-fold higher in male mice than in female mice. On the contrary, the proportion of ODC present as a complex with antizyme was 24-fold higher in females than in males, and the decay of renal ODC activity after cycloheximide treatment was about 5-fold more rapid in females than in males. Administration of testosterone to female mice, a procedure known to prolong the half-life of renal ODC, increased both ODC activity and the content of ODC-antizyme complex, but decreased the antizyme/ODC ratio in the kidney. These results are consistent with the previous observation in HTC cells that the decay rate of ODC activity in the presence of cycloheximide correlated well with the proportion of ODC present as a complex with antizyme, suggesting the ubiquitous role of antizyme in ODC degradation.     

Ornithine decarboxylase antizyme inhibitor 2 regulates intracellular vesicle trafficking

Antizyme inhibitor 1 (AZIN1) and 2 (AZIN2) are proteins that activate ornithine decarboxylase (ODC), the key enzyme of polyamine biosynthesis. Both AZINs release ODC from its inactive complex with antizyme (AZ), leading to formation of the catalytically active ODC. The ubiquitously expressed AZIN1 is involved in cell proliferation and transformation whereas the role of the recently found AZIN2 in cellular functions is unknown. Here we report the intracellular localization of AZIN2 and present novel evidence indicating that it acts as a regulator of vesicle trafficking. We used immunostaining to demonstrate that both endogenous and FLAG-tagged AZIN2 localize to post-Golgi vesicles of the secretory pathway. Immuno-electron microscopy revealed that the vesicles associate mainly with the trans-Golgi network (TGN). RNAi-mediated knockdown of AZIN2 or depletion of cellular polyamines caused selective fragmentation of the TGN and retarded the exocytotic release of vesicular stomatitis virus glycoprotein. Exogenous addition of polyamines normalized the morphological changes and reversed the inhibition of protein secretion. Our findings demonstrate that AZIN2 regulates the transport of secretory vesicles by locally activating ODC and polyamine biosynthesis.     

Inducible expression of antizyme 1 in prostate cancer cell lines after lentivirus mediated gene transfer

The prostate has the highest level of polyamines among all tissues, and it is the only tissue in which polyamines are purposely synthesized for export. It has been suggested that the high local polyamine concentrations suppress cell growth of primary prostatic carcinomas and that this growth control is lost when cancer cells metastasize. It has also been shown that the sensitivity to polyamine-induced growth arrest correlates with antizyme induction in prostate carcinoma cell lines. In this study, we evaluated the sensitivity of poorly metastatic (LNCaP) and highly metastatic (DU145) prostate cancer cell lines to conditional antizyme 1 over-expression. Antizyme 1 induction resulted in a marked loss of ODC activity and polyamine uptake in both cell lines. However, the proliferation of LNCaP cells was repressed by antizyme 1 induction, whereas the proliferation of DU 145 cells was not affected. Neither cell line showed any reduction in polyamine pools after manipulation nor did polyamine addition into the medium save the LNCaP cells from the growth retardation. The growth inhibition of LNCaP cells was accompanied by accumulation of cells in the G1 phase and depletion of cyclin E1 protein. These results confirm that different prostate cancer cell lines show diverse sensitivities to antizyme 1 which may not be directly polyamine related. The high gene transfer capacity of the used lentiviral vector makes the present approach a useful tool to study the therapeutic potential of antizyme 1 both in cell cultures and experimental animals.     

Ornithine decarboxylase-antizyme is rapidly degraded through a mechanism that requires functional ubiquitin-dependent proteolytic activity.

Antizyme is a polyamine-induced cellular protein that binds to ornithine decarboxylase (ODC), and targets it to rapid ubiquitin-independent degradation by the 26S proteasome. However, the metabolic fate of antizyme is not clear. We have tested the stability of antizyme in mammalian cells. In contrast with previous studies demonstrating stability in vitro in a reticulocyte lysate-based degradation system, in cells antizyme is rapidly degraded and this degradation is inhibited by specific proteasome inhibitors. While the degradation of ODC is stimulated by the presence of cotransfected antizyme, degradation of antizyme seems to be independent of ODC, suggesting that antizyme degradation does not occur while presenting ODC to the 26S proteasome. Interestingly, both species of antizyme, which represent initiation at two in-frame initiation codons, are rapidly degraded. The degradation of both antizyme proteins is inhibited in ts20 cells containing a thermosensitive ubiquitin-activating enzyme, E1. Therefore we conclude that in contrast with ubiquitin-independent degradation of ODC, degradation of antizyme requires a functional ubiquitin system.     

Recurrent Emergence of Catalytically Inactive Ornithine Decarboxylase Homologous Forms That Likely Have Regulatory Function

Ornithine decarboxylase (ODC) catalyzes the first and rate limiting step in the biosynthesis of polyamines in most eukaryotes. Because polyamines have pleiotropic and often dramatic effects on cellular processes at both high and low concentrations, ODC expression is tightly controlled. ODC is regulated by a family of polyamine-induced proteins, antizymes, which bind to, and inactivate it. In mammals, and apparently most vertebrates, antizymes are in turn antagonized by proteins called antizyme inhibitors. Antizyme inhibitors are homologs of ODC that have lost their decarboxylation activity but have retained their ability to bind antizyme, in most cases even more tightly than ODC. We present a phylogenetic analysis of over 200 eukaryotic homologs of ODC and evaluate their potential to be either true ODCs or catalytically inactive proteins that might be analogs of the previously identified antizyme inhibitors. This analysis yielded several orthologous groups of putative novel antizyme inhibitors each apparently arising independently. In the process we also identify previously unrecognized ODC paralogs in several evolutionary branches, including a previously unrecognized ODC paralog in mammals, and we evaluate their biochemical potential based on their pattern of amino acid conservation.