CELL SURFACE ALTERATIONS BY TAXOL ASSOCIATED WITH ABNORMAL MORPHOGENESIS IN THE CHICK EMBRYO

The anticancer drug taxol brings about its biological effects by altering the stability of microtubules. We have examined the effects of taxol on early morphogenesis in chick embryos culturedin vitro. Taxol induced various abnormalities in the developing nervous system, heart and somites as well as general retardation of development. SEM studies revealed that taxol treatment leads to dramatic alterations in the embryonic cell surfaces. Time-course experiments demonstrated that the action of taxol is very rapid and becomes evident within a few minutes at the ultrastructural level. Taxol thus throws embryonic cell adhesion and motility out of balance. This appears to be the major cause of abnormal morphogenesis in taxol-treated embryos.     

In vitro culture and precocious germination ofTaxus embryos

Summary The lengthy dormancy requirement of yew seeds can be overcome with a simple in vitro method. Viable embryos were excised from seeds ofTaxus brevifolia and four cultivars ofT. media over a range of developmental stages. Embryos were cultured in several basal media formulations (Whites’, Gamborg’s B5 and Murashige and Skoog’s) under dark or light. After a lag period of 1 to 2 wk, embryos of both species germinated precociously. Germination rates of up to 70% were obtained withT. media cv. Hicksi embryos. The highest rates of germination were obtained in White’s and MS media. Embryos excised from green seeds with undeveloped arils showed the highest germination rates. As the seeds approached maturity, in vitro germination rates of the excised embryos declined dramatically. Green seeds and seeds with developing arils could be stored at 5° C without large loss in embryo germination. Seeds with fully developed arils could be stored frozen at −20° C for 1 wk while still allowing about 50% of embryo germination. At least 30% of the precociously germinated embryos of both species were able to develop into full seedlings. Our method appears to be generally applicable toTaxus spp. This research was supported by a grant from the Hawaii Biotechnology Group, Inc.     

Mitotic drug targets

Mitosis is the key event of the cell cycle during which the sister chromatids are segregated onto two daughter cells. It is well established that abrogation of the normal mitotic progression is a highly efficient concept for anti‐cancer treatment. In fact, various drugs that target microtubules and thus interfere with the function of the mitotic spindle are in clinical use for the treatment of various human malignancies for many years. However, since microtubule inhibitors not only target proliferating cells severe side effects limit their use. Therefore, the identification of novel mitotic drug targets other than microtubules have gained recently much attention. This review will summarize the latest developments on the identification and clinical evaluation of novel mitotic drug targets and will introduce novel concepts for chemotherapy that are based on recent progress in our understanding how mitotic progression is regulated and how anti‐mitotic drugs induce tumor cell death. J. Cell. Biochem. 111: 258–265, 2010. © 2010 Wiley‐Liss, Inc.     

TRAIL Receptors Serve as Stress-Associated Molecular Patterns to Promote ER-Stress-Induced Inflammation

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基于定量PCR技术探讨紫杉醇生物合成的限速步骤

次生代谢产物牛物合成受到发育和诱导的调控,本实验研究了组织分化和诱导处理对紫杉醇生物合成的影响,并采用定量PCR技术分析了紫杉醇生物合成不同阶段关键酶基因的动态表达特征。结果表明。紫杉醇主要分布在中国红豆杉（Taxus chinensis）树皮和根皮组织中,针叶内含量很少,催化紫杉醇功能官能团连接的关键酶摹因也主要定位在树皮和根皮组织巾;茉莉酸甲酯（MJ）和真菌诱导子F5分别提高了中国红豆杉悬浮培养细胞HG-1紫杉醇得率8倍和10倍,同时有效诱导紫杉醇生物合成基因的表达。发现催化紫杉醇侧链连接的基因与紫杉醇生物合成早正相关。结果表明。紫杉醇生物合成的限速步骤是催化功能官能团连接的步骤。     

植物细胞工程技术生产紫杉醇研究进展

紫杉醇是一种二萜类生物碱,主要从红豆杉树皮中分离提取。目前采用植物细胞工程技术是提高紫杉醇产率、保护稀缺资源红豆杉、解决紫杉醇药源紧缺的一种最有效方法。该文对近十年来国内外有关采用植物细胞工程技术生产紫杉醇的研究进展,包括红豆杉细胞系的建立、悬浮培养条件的研究、生物反应器培养以及紫杉醇合成代谢调控方面的最新研究进展进行综述。并重点论述了前体、诱导子和代谢支路抑制剂对红豆杉细胞悬浮培养生产紫杉醇的影响,为植物细胞工程技术生产紫杉醇提供借鉴和参考。     

Discovery of facile amides-functionalized rhodanine-3-acetic acid derivatives as potential anticancer agents by disrupting microtubule dynamics

Microtubule dynamics are crucial for multiple cell functions, and cancer cells are particularly sensitive to microtubule-modulating agents. Here, we describe the design and synthesis of a series of (Z)-2-(5-benzylidene-4-oxo-2-thioxothiazolidin-3-yl)-N-phenylacetamide derivatives and evaluation of their microtubule-modulating and anticancer activities in vitro. Proliferation assays identified I₂₀ as the most potent of the antiproliferative compounds, with 50% inhibitory concentrations ranging from 7.0 to 20.3 µM with A549, PC-3, and HepG2 human cancer cell lines. Compound I₂₀ also disrupted cancer A549 cell migration in a concentration-dependent manner. Immunofluorescence microscopy, transmission electron microscopy, and tubulin polymerisation assays suggested that compound I₂₀ promoted protofilament assembly. In support of this possibility, computational docking studies revealed a strong interaction between compound I₂₀ and tubulin Arg β369, which is also the binding site for the anticancer drug Taxol. Our results suggest that (Z)-2-(5-benzylidene-4-oxo-2-thioxothiazolidin-3-yl)-N-phenylacetamide derivatives could have utility for the development of microtubule-stabilising therapeutic agents.     

前体及诱导子和发酵条件对烟曲霉TMS-26产紫杉醇发酵体系优化

紫杉醇是一种广谱的抗癌药物,因其具有独特的抗癌机制、良好的抗癌效果和供不应求的市场等特征而备受关注。紫杉醇具有重大经济效益,但产量受到制约,价格极为昂贵,通过内生真菌发酵法生产紫杉醇能在一定程度上缓解其来源困难的问题。在产紫杉醇内生真菌TMS-26发酵液中添加前体物质和诱导子,并通过对接种量、装液量、初始pH和发酵时间等条件进行优化研究。单因素及正交试验表明在PDB培养基中加入苯丙氨酸20mg/L、苯甲酸钠30mg/L、乙酸钠8g/L、甘氨酸15mg/L、CuSO₄ 0.05mg/L、H₂O₂ 6mmol/L、3,5-二硝基水杨酸15mg/L时能有效提高紫杉醇产量,比优化前增产46.64%,达到446.28µg/L,并且发现最适菌株TMS-26的发酵条件为pH7.5、接种量5%、装液量120mL/250mL、发酵时间为10d。     

紫杉醇免疫检测方法的研究进展

紫杉醇是一种有效的抗肿瘤药物,广泛应用于治疗卵巢癌、乳腺癌和肺癌等癌症。紫杉醇在紫杉树皮中的含量极低（仅为0．01％）,而且紫杉醇是一种对蛋白质有着高亲和力的小分子,在体液中约有98％的分子与蛋白质结合,因此需要一种高灵敏度、高通量的检测方法对紫杉醇进行鉴定。在分析紫杉醇检测方法的基础之上,综述了紫杉醇免疫学检测方法的研究进展,包括紫杉醇半抗原的分子修饰、蛋白偶联物的构建和鉴定以及免疫学检测方法在植物组织和病人血浆中紫杉醇定性和定量中的应用。