Characterization of a membrane-associated protein kinase of multidrug-resistant HL60 cells which phosphorylates P-glycoprotein

Cells containing increased levels of the membrane phosphoprotein P-glycoprotein exhibit a multidrug-resistant phenotype. In the present study we have analyzed protein kinases capable of phosphorylating P-glycoprotein in membranes of HL60 cells isolated for resistance to vincristine. Analysis of this system demonstrates that in isolated membranes the protein kinase inhibitor staurosporine greatly reduces P-glycoprotein phosphorylation. In contrast, the kinase inhibitor H-7 does not affect this reaction. Fractionation of solubilized membrane proteins from sensitive and resistant cells on DEAE-cellulose reveals a major protein kinase (PK-1) which exhibits optimal activity in the presence of Mn2+ and histone H1. This enzyme fraction does not contain detectable levels of protein kinase C or cAMP-dependent protein kinase. PK-1 phosphorylation of two endogenous proteins is, however, greatly enhanced in the presence of phosphatidylserine or phosphatidyl-inositol. In reaction mixtures containing Mg2+ or Mn2+ in the absence of phospholipid, PK-1 from resistant cells phosphorylates an endogenous protein of 180 kilodaltons (P180), which exhibits an electrophoretic mobility identical to P-glycoprotein. In parallel experiments with PK-1 from sensitive cells there is no detectable phosphorylation of a P180 protein. P180 phosphorylated by PK-1 from resistant cells is immunoprecipitated by antibody against P-glycoprotein. Additional studies demonstrate that PK-1 is capable of phosphorylating specific synthetic peptides which correspond to the sequence of P-glycoprotein. Peptide phosphorylation occurs at both serine and threonine residues. These studies thus identify a novel membrane-associated protein kinase in HL60 cells which is capable of phosphorylating P-glycoprotein. This enzyme may have an important role in regulating levels of multidrug resistance.     

Adriamycin resistance in HL60 cells in the absence of detectable P-glycoprotein

Previous studies have shown that the development of multi-drug resistance in cell lines treated with chemotherapeutic agents is closely associated with the overexpression of a 170-180 kilodalton surface membrane glycoprotein (P-glycoprotein). In the present study a monoclonal antibody against the P-glycoprotein was used to determine if this protein is overexpressed in multi-drug resistant HL60 cells. Using either indirect immunofluorescent staining or immunoblot analysis P-glycoprotein could not be detected in HL60 cells isolated for resistance to adriamycin. In contrast HL60 cells isolated for resistance to vincristine contain the P-glycoprotein and the amount of this material increases with increasing levels of resistance. These studies thus demonstrate adriamycin resistance in P-glycoprotein negative HL60 cells. Furthermore adriamycin and vincristine are found to have distinct effects in inducing overexpression of P-glycoprotein in the HL60 cell line. This information could be useful in the development of therapeutic strategies for the treatment of certain forms of cancer.     

松辽盆地阿尔必期微体浮游植物新属种

该文描述了松辽盆地中白垩世阿尔必期泉头组三段的微体浮游植物化石1新属7新种,隶属于微咸水沟鞭藻类2属5新种(含4新亚种),淡水绿藻1属1种和疑源类1新属1新种。并对沟鞭藻 Ngktericysta Bint,1986进行了修订。     

松辽盆地白垩纪微体浮游植物群及其环境讨论

该文报道了松辽盆地白垩纪丰富的非海相微体浮游植物群,主要是沟鞭藻类及一些绿藻和疑源类;论述了藻类的生物地层特征,自下而上初步划分出10个组合带;结合微量元素和古地磁等资料,较详细地讨论了含微体浮游植物组段的沉积环境,认为松辽盆地在白垩纪至少遭受过两次重要的海侵(分别在青山口组一段及嫩江组一、二段沉积时期),导致古松辽湖泊五种不同水体环境的演替,指出微体浮游植物组合的变化是受古盐度、古温度和古水深等因素控制的。此外,对有关组段的地质时代也进行了讨论,进一步补充了新的浮游植物化石证据。     

Generation of, lipid neutrophil chemoattractant activity by histamine-stimulated cultured endothelial cells

Endothelial cell-neutrophil interactions are an important aspect of inflammatory responses. Because vascular endothelial cells respond to the inflammatory mediator histamine, these studies determined whether histamine could induce endothelial cells to release substances that affect human neutrophil migration. Cultured bovine and human endothelial cells incubated with histamine released neutrophil chemoattractant activity within 1 min; peak levels were noted in 45 min. Cimetidine, an H2 receptor antagonist, blocked chemoattractant production, whereas diphenhydramine, an H1 receptor antagonist, did not. Cycloheximide did not inhibit release of chemoattractant activity, suggesting de novo protein synthesis was not necessary for its appearance. Extraction with acidified diethyl ether partitioned all neutrophil chemoattractant activity into the organic phase. The lipoxygenase pathway inhibitors, diethylcarbamazine and 5,8,11,14 eicosatetraynoic acid, inhibited generation of this lipophilic chemoattractant activity, whereas indomethacin, a cyclo-oxygenase inhibitor, did not. Resolution of the histamine-induced endothelial cell-derived chemoattractant activity by reverse-phase high pressure liquid chromatography yielded several peaks of chemoattractant activity, none of which co-eluted with leukotriene B4, platelet-activating factor, or two mono-hydroxyeicostetraenoic acids. These findings suggest that endothelial cells release lipid neutrophil chemoattractant activity that may play a role in inflammatory responses associated with histamine.     

应用X光荧光法检测硫的化学位移和研究病毒感染植物中硫状态的改变

应用双晶X光荧光光谱仪研究生理性和病理性植物组织中某些元素的分布特征已有报道。本研究工作中采用日本国东芝厂出品的AFV701型双晶X光荧光光谱仪,测定并比较了植物感染病毒前、后的叶片中,不同硫状态相对比例的改变。应用此仪器首先测定各种标准硫化物硫状态的化学位移数值,在确定标准图谱及可分性之后,再测定感染蚕豆萎蔫病毒(BBWV)、黄瓜花叶病毒(CMV)、芜菁花叶病毒(TuMV)的茄、苋色藜、烟草和萝卜等四种寄主植物的显症病叶中不同硫状态的相对比例。发现应用此技术能明显地区别出病态植物与健康植物中硫状态相对比例的变化差异。     

Construction and packaging of pseudotype retrovirus containing human N—ras cDNA antisense sequence and its biological effects on human hepatoma cells

N-ras is one of the transforming genes in human hepatic cancer cells.It has been found that N-ras was overexpressed at the mRNA and protein level in hepatoma cells.In order to explore the biological roles of N-ras in human hepatic carcinogenesis and the potential application in control of cancer cell growth,a preudotype retrovirus containing antisense sequence of human N-ras was constructed and packaged.A recombinant retrovirus vector containing antisense or sense sequences of N-ras cDNA was constructed by pZIP-NeoSV(X)1.The pseudotype virus was packaged ang rescued by transfection and infection in PA317 and ψ 2 helper cells.It has been demonstrated that the pseudotype retrovirus containing antisense N-ras sequence did inhibit the growth of human PLC/PRF/5 hepatoma cells accompanied with inhibition of p21 expression,while the retrovirus containing sense sequence had none.The pseudotype virus had no effect on human diploid fibroblasts.     

天麻消化紫萁小菇及蜜环菌过程中细胞超微结构变化的研究

本文对天麻种子消化入侵的紫萁小菇菌丝及营养繁殖茎消化蜜环菌过程中,细胞超微结构的变化进行了研究。观察结果表明:紫萁小菇侵入种胚后,染菌胚细胞的细胞器逐渐消失,其细胞质产生囊状体起消化菌丝的作用,存在于胚细胞中的紫萁小菇菌丝有脱壁或失去细胞质成为空腔等变化;种子萌发形成的原球茎消化紫萁小菇的方式同胚萌发阶段类同。蜜环菌侵入原球茎分化的营养繁殖茎后,皮层细胞产生消化酶类颗粒或囊状体包围并分解蜜环菌菌丝;被皮层细胞消化的菌丝残物或部分菌丝进入皮层内面的大型细胞,此时大型细胞的代谢功能显著增强,该细胞中的各种水解酶颗粒及液泡等完成对菌体物质的最终同化。紫萁小菇及蜜环菌先后在天麻有性繁殖和无性繁殖阶段侵染供给其营养,但菌丝被消化过程中的细胞形态变化、被消化方式不完全一致。