应力、应力纤维、Rho三者关系的研究进展

应力影响细胞的形状和骨架结构并因此控制许多与组织发展有关的关键行为，因此应力作用下细胞的种种响应问题成为生物力学目前最热的研究领域，应力纤维、Rho在研究应力信号如何被细胞感受过程中占据核心地位并已成为研究者关注的重点。本文依据国内外近几年进行的细胞对应力响应的研究进展对应力、应力纤维、Rho三者的相互关系进行了综述。     

乳腺癌PKCs表达与其分化程度的相关性

为了研究蛋白激酶C（PKC）和乳腺癌分化及转移的相关性，采用免疫组化SABC方法检测48例人乳腺癌的PKC蛋白表达，包括PKCα、βI、η和ξ等蛋白。结果表明，与分化低的乳腺癌对比，分化高的乳腺癌PKC蛋白表达水平提高。在临床I－II和III级的乳腺癌中，PKCη蛋白表达率分别为61．5％和9．1％，具有显著意义（P＜0．05），PKCξ蛋白表达率分别为92．3％和27．3％，具有极显著意义（P＜0．01）。另外，在乳腺癌未转移的病例中，PKCs蛋白表达水平普遍较高，其中PKCξ与肿转移相关性极显著（P＜0．01）。此外，PKCs蛋白表达水平与肿瘤患者年龄无关。以上结果提示，PKCη和PKCξ有可能作为评价临床乳腺癌分化和转移程度的指标之一。     

Changes in glycosaminoglycan sulfation and protein kinase C subcellular distribution during differentiation of the human colon tumor cell line Caco-2

Summary During the spontaneous differentiation (day 5 to day 15 of the culture) of Caco-2 cells, the sulfation of cell layer glycosaminoglycans increased, whereas protein kinase C activity was concomitantly redistributed from the membrane to the cytosol. The protein kinase C activators, 4-phorbol 12-myristate, 13-acetate and 1,2-dioctanoyl-glycerol inhibited glycosaminoglycan sulfation. By contrast, 4-phorbol 12, 13 didecanoate was ineffective.These results suggest that membrane-bound PKC may exert a modulatory effect on glycosaminoglycan sulfation, and this effect is gradually attenuated as Caco-2 cell differentiation progresses.     

Effect of protein kinase C activation and depletion on insulin stimulation of glycogen synthesis in cultured hepatoma cells

Summary Insulin stimulation of glycogen synthesis was nearly abolished in hepatoma cells shortly treated with 4 ß-phorbol 12 \-myristate, 13 -acetate (protein kinase C activation) but remained unmodified in cells chronically treated with the phorbol ester (protein kinase C depletion). Thus, although exogenous activation of protein kinase C results in an inhibition of insulin action, protein kinase C depletion has no influence on this process. The results suggest that, in hepatoma cells, no endogenous activation of protein kinase C may occur in response to the signal triggered by insulin.     

用复性电泳技术研究溶酶体蛋白水解酶的性质和活性

介绍了复性单向电泳（Ｇ－ＰＡＧＥ）和复性双向电泳（ＩＥＦ－Ｇ－ＰＡＧＥ）的概念、原理、方法、应用范围、优缺点和使用该方法应注意的事项。并用该方法定性和定量分析了Ｃ６大白鼠神经胶质瘤细胞酸性溶酶体蛋白水解酶的性质和活性。     

腺病毒载体介导胸苷激酶基因/ACV对大鼠脑胶质瘤的离体和活体杀伤作用

建立缺陷型重组腺病毒载体介导的胸苷激酶基因治疗系统（Adtk／ACV）,进行离休和活体治疗大鼠C6脑胶质瘤研究．将质粒pAdtk与pJM17共转染腺病毒包装细胞──293细胞,同源重组后制备纯化的Adtk并以PCR证实;Adtk对离休C6细胞的杀伤作用随着Adtk滴度和ACV剂量增加而增强,并有旁观者效应,而未转染C6细胞和AdLacZ转染C6细胞均未被杀伤;扫描电镜下见Adtk／ACV处理的细胞呈明显病理改变;ACV可有效杀伤Adtk／C6细胞．在大鼠脑立体定向仪引导下于右额叶接种2×106个C6细胞,分别于第3,6,8和10天原位注射Adtk,同时腹腔注射ACV（100mg／（kg·d-1））,3d组、6d组、8d组大鼠成活期在90d以上,组织学检查未发现肿瘤细胞．10d组成活期（28．5±4．6）d,而C6未治疗组和AdLacZ／ACV治疗组成活期分别为（1．8±3．1）d和（14．0±2．2）d．本文建立的Adtk／ACV在离体和活体水平对大鼠C6脑胶质瘤有强烈的杀伤作用,效果确实,应用方便,ACV价格便宜,有潜在临床应用价值．     

细胞分裂素对大豆叶片衰老过程中蛋白激酶基因表达的影响

利用真核生物催化丝氨酸／苏氨酸残基磷酸化的蛋白激酶ＶＩ和Ⅷ保守区设计兼并引物,进行ＲＴ－ＰＣＲ扩增反应,研究大豆叶片衰老过程中蛋白激酶基因表达的变化及外源细胞分裂素处理的作用。结果表明人工合成的细胞分裂素－６ＢＡ预处理很可能在转录水平上正向调节一些蛋白激酶基因的表达,而诱导衰老处理则起负调控作用。     

Roles of protein kinase C in oocyte meiotic maturation and fertilization

Protein kinase C (PKC) is a superfamily of Ser/Thr protein kinases that is distributed widely in eukaryotes. It plays key regulatory roles at multiple steps of oocyte meiotic maturation and fertilization. During the process of meiotic maturation, the activation of PKC in cumulus cells stimulates meiotic maturation, whereas the activation of PKC in oocytes results in the inhibition of germinal vesicle breakdown. PKC activity increases following the meiotic maturation, and decreases at the transition of metaphase/anaphase in meiosis I, so as to facilitate the release of the first polar body and the entry of meiosis II. In fertilization of mammalian oocytes, PKC may act as one of the downstream targets of Ca2+ to stimulate the cortical granule exocytosis, release the oocytes from MII arrest and to induce pronucleus formation. PKC is also involved in the regulation of maturation promoting factor (MPF) and mitogen-activated protein kinase (MAPK). Several PKC isoforms have been identified in mammalian oocytes, and there is evidence showing that classical PKCs may be the principal mediator of oocyte cortical reaction.     

纳豆激酶基因克隆及其在大肠杆菌中活性表达研究

以纳豆芽孢杆菌基因组DNA为模板，PCR扩增了纳豆激酶基因（natto kinase gene）中编码前肽、成熟肽的核苷酸序列（pro-NK），构建大肠杆菌表达质粒pTYB102，转化大肠杆菌ER2566。在IPTG诱导下，分别在15℃（14h）、30℃（3h）、37℃（2h）条件下培养，pTYB102均能表达出有活性的纳豆激酶。实验证实纳豆激酶基因得到活性表达需要Pro序列。SDS－PAGE表明，15℃和30℃和37℃培养表达的杂蛋白更少。薄层扫描测定表达的纳豆激酶占菌体总蛋白30％以上。     

乳酸对肌酸激酶活性抑制作用的研究

应用酶活测定的方法分析了肌酸激酶在不同浓度乳酸中的活性变化．实验结果说明在失活过程中没有蛋白质的聚沉；肌酸激酶的失活遵循一级反应的一相过程；失活与乳酸浓度和作用时间呈正相关．通过对比实验结果显示，乳酸是通过其解离状态的H^ 改变了机体PH值而影响肌酸激酶活性的，并且就乳酸对肌酸激酶影响及其意义进行探讨．