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181.
Erk is involved in the differentiation induced by diallyl disulfide in the human gastric cancer cell line MGC803 总被引:3,自引:0,他引:3
Ling H Zhang LY Su Q Song Y Luo ZY Zhou XT Zeng X He J Tan H Yuan JP 《Cellular & molecular biology letters》2006,11(3):408-423
Diallyl disulfide (DADS) is a major constituent of garlic. Previously, we found that DADS both inhibited proliferation in
human gastric cancer cells in vitro and in vivo, and induced G2/M arrest. In this study, we investigated whether this differentiation effect was induced by DADS in human
gastric cancer MGC803 cells, and whether it was related to an alteration in ERK activity. The results showed that the growth
of MGC803 cells was inhibited by DADS. Cells treated with DADS displayed a lower nucleocytoplasmic ratio and tended to form
gland and intercellular conjunction structures. The ConA-mediated cell agglutination ratio and cells’ ALP specific activity
decreased. In MGC803 cells, dye transfer was limited to a few cells neighbouring the dye-injected cell and to a depth of 1–2
layers beneath the scrape site. However, after treatment with DADS, the LY (Lucifer Yellow) was transferred to several cells
immediately neighbouring the microinjected cell and to a depth of 2–4 cell layers from the scrape site. This indicated that
DADS induced differentiation in MGC803 cells. Western blot analysis revealed that although DADS did not influence the quantity
of ERK1/2 protein expressed, it did decrease its phosphorylation in a concentration-dependent manner, compared with the controls.
At 30 mg·L−1, DADS inhibited the activation of ERK1/2 in 15–30 min. These results suggested that the DADS-induced differentiation of MGC803
cells involved an alteration of the ERK1/2 signaling pathway. 相似文献
182.
《MABS-AUSTIN》2013,5(8):1358-1366
ABSTRACTReversed-phase liquid chromatography (RPLC) separations of proteins using optical detection generally use trifluoroacetic acid (TFA) because it is a strong, hydrophobic acid and a very effective ion-pairing agent for minimizing chromatographic secondary interactions. Conversely and in order to avoid ion suppression, analyses entailing mass spectrometry (MS) detection is often performed with a weaker ion-pairing modifier, like formic acid (FA), but resolution quality may be reduced. To gain both the chromatographic advantages of TFA and the enhanced MS sensitivity of FA, we explored the use of an alternative acid, difluoroacetic acid (DFA). This acid modifier is less acidic and less hydrophobic than TFA and is believed to advantageously affect the surface tension of electrospray droplets. Thus, it is possible to increase MS sensitivity threefold by replacing TFA with DFA. Moreover, we have observed DFA ion pairing to concomitantly produce higher chromatographic resolution than FA and even TFA. For this reason, we prepared and used MS-quality DFA in place of FA and TFA in separations involving IdeS digested, reduced NIST mAb and a proprietary antibody-drug conjugate (ADC), aiming to increase sensitivity, resolution and protein recovery. The resulting method using DFA was qualified and applied to two other ADCs and gave heightened sensitivity, resolution and protein recovery versus analyses using TFA. This new method, based on a purified, trace metal free DFA, can potentially become a state-of-the-art liquid chromatography-MS technique for the deep characterization of ADCs. 相似文献
183.
目的:通过二烯丙基二硫诱导白血病K562细胞发生自噬性死亡,探讨其作用机制。方法:40 mg/LDADS作用K562细胞12小时后,透射电镜观察K562细胞超微结构,MDC染色荧光显微镜观察自噬泡及流式细胞仪定量检测自噬率,RT-PCR检测Beclin1mRNA的表达水平。结果:DADS作用后的K562细胞后,透射电镜可观察到胞质内出现大量自噬体;MDC染色荧光显微镜观察显示,K562细胞胞浆中的自噬泡明显增多,而空白组与溶媒组胞浆中的自噬泡很少;流式细胞术定量测定空白对照组、溶媒对照组、DADS药物组自噬率分别为(7.27±5.60)%、(7.10±5.13)%、(27.39±6.51)%(P〈0.05);空白对照组为0.658±0.007,溶媒对照组为0.671±0.012,两者的Beclin1mRNA的表达强度无明显差异(P〉0.05),DADS药物组为0.911±0.008,高于对照组(P〈0.05)。结论:二烯丙基二硫可诱导白血病k562细胞发生自噬性死亡,其机制可能与Beclin1的上调有关。 相似文献
184.
Molecular cloning,expression, and hormonal regulation of the chicken microsomal triglyceride transfer protein 总被引:1,自引:0,他引:1
N. Erwin Ivessa Edward Rehberg Bernadette Kienzle Fridolin Seif Robert Hermann Marcela Hermann Wolfgang J. Schneider David A. Gordon 《Gene》2013
During an egg-laying cycle, oviparous animals transfer massive amounts of triglycerides, the major lipid component of very low density lipoprotein (VLDL), from the liver to the developing oocytes. A major stimulus for this process is the rise in estrogen associated with the onset of an egg-laying cycle. In mammals, the microsomal triglyceride transfer protein (MTP) is required for VLDL assembly and secretion. To enable studies to determine if MTP plays a role in basal and estrogen-stimulated VLDL assembly and secretion in an oviparous vertebrate, we have cloned and sequenced the chicken MTP cDNA. This cDNA encodes a protein of 893 amino acids with an N-terminal signal sequence. The primary sequence of chicken MTP is, on average, 65% identical to that of mammalian homologs, and 23% identical to the Drosophila melanogaster protein. We have obtained a clone of chicken embryo fibroblast cells that stably express the avian MTP cDNA and show that these cells display MTP activity as measured by the transfer of a fluorescently labeled neutral lipid. As in mammals, chicken MTP is localized to the endoplasmic reticulum as revealed by indirect immunofluorescence and by the fact that its N-linked oligosaccharide moiety remains sensitive to endoglycosidase H. Endogenous, enzymatically active MTP is also expressed in an estrogen receptor-expressing chicken hepatoma cell line that secretes apolipoprotein B-containing lipoproteins. In this cell line and in vivo, the expression and activity of MTP are not influenced by estrogen. Therefore, up-regulation of MTP in the liver is not required for the increased VLDL assembly during egg production in the chicken. This indicates that MTP is not rate-limiting, even for the massive estrogen-induced secretion of VLDL accompanying an egg-laying cycle. 相似文献
185.
186.
Katariina M. Hutterer Robert W. Hong Jonathon Lull Xiaoyang Zhao Tian Wang Rex Pei M. Eleanor Le Oleg Borisov Rob Piper Yaoqing Diana Liu Krista Petty Izydor Apostol Gregory C. Flynn 《MABS-AUSTIN》2013,5(4):608-613
Manufacturing-induced disulfide reduction has recently been reported for monoclonal human immunoglobulin gamma (IgG) antibodies, a widely used modality in the biopharmaceutical industry. This effect has been tied to components of the intracellular thioredoxin reduction system that are released upon cell breakage. Here, we describe the effect of process parameters and intrinsic molecule properties on the extent of reduction. Material taken from cell cultures at the end of production displayed large variations in the extent of antibody reduction between different products, including no reduction, when subjected to the same reduction-promoting harvest conditions. Additionally, in a reconstituted model in which process variables could be isolated from product properties, we found that antibody reduction was dependent on the cell line (clone) and cell culture process. A bench-scale model using a thioredoxin/thioredoxin reductase regeneration system revealed that reduction susceptibility depended on not only antibody class but also light chain type; the model further demonstrates that the trend in reducibility was identical to DTT reduction sensitivity following the order IgG1λ > IgG1κ > IgG2λ > IgG2κ. Thus, both product attributes and process parameters contribute to the extent of antibody reduction during production. 相似文献
187.
Mariapia Vairetti Andrea Ferrigno Vittoria RizzoGiulia Ambrosi Alberto BianchiPlinio Richelmi Fabio BlandiniMarie-Therese Armentero 《生物化学与生物物理学报:疾病的分子基础》2012,1822(2):176-184
In Parkinson's disease (PD), aside from the central lesion, involvement of visceral organs has been proposed as part of the complex clinical picture of the disease. The issue is still poorly understood and relatively unexplored. In this study we used a classic rodent model of nigrostriatal degeneration, induced by the intrastriatal injection of 6-hydroxydopamine (6-OHDA), to investigate whether and how a PD-like central dopaminergic denervation may influence hepatic functions. Rats received an intrastriatal injection of 6-OHDA or saline (sham), and blood, cerebrospinal fluid, liver and brain samples were obtained for up to 8 weeks after surgery. Specimens were analyzed for changes in cytokine and thyroid hormone levels, as well as liver mitochondrial alterations. Hepatic mitochondria isolated from animals bearing extended nigrostriatal lesion displayed increased ROS production, while membrane potential (ΔΨ) and ATP production were significantly decreased. Reduced ATP production correlated with nigral neuronal loss. Thyroid hormone levels were significantly increased in serum of PD rats compared to sham animals while steady expression of selected cytokines was detected in all groups. Hepatic enzyme functions were comparable in all animals. Our study indicates for the first time that in a rodent model of PD, hepatic mitochondria dysfunctions arise as a consequence of nigrostriatal degeneration, and that thyroid hormone represents a key interface in this CNS-liver interaction. Liver plays a fundamental detoxifying function and a better understanding of PD-related hepatic mitochondrial alterations, which might further promote neurodegeneration, may represent an important step for the development of novel therapeutic strategies. 相似文献
188.
Over the last seven years our laboratory has focused on the determination of the structural aspects of nucleoside triphosphate diphosphohydrolases (NTPDases) using site-directed mutagenesis and computational comparative protein modeling to generate hypotheses and models for the hydrolytic site and enzymatic mechanism of the family of NTPDase nucleotidases. This review summarizes these studies utilizing NTPDase3 (also known as CD39L3 and HB6), an NTPDase family member that is intermediate in its characteristics between the more widely distributed and studied NTPDase1 (also known as CD39) and NTPDase2 (also known as CD39L1 and ecto-ATPase) enzymes. Relevant site-directed mutagenesis studies of other NTPDases are also discussed and compared to NTPDase3 results. It is anticipated that many of the results and conclusions reached via studies of NTPDase3 will be relevant to understanding the structure and enzymatic mechanism of all the cell-surface members of this family (NTPDase1–3, 8), and that understanding these NTPDase enzymes will aid in modulating the many varied processes under purinergic signaling control. This review also integrates the site-directed mutagenesis results with a recent 3-D structural model for the extracellular portion of NTPDases that helps explain the importance of the apyrase conserved regions (ACRs) of the NTPDases. Utilizing this model and published work from Dr Guidotti's laboratory concerning the importance and characteristics of the two transmembrane helices and their movements in response to substrate, we present a speculative cartoon model of the enzymatic mechanism of the membrane-bound NTPDases that integrates movements of the extracellular region required for catalysis with movements of the N- and C-terminal transmembrane helices that are important for control and modulation of enzyme activity. 相似文献
189.
Justin M. Whiteley Simon Hafner Sang Sub Han Seul Cham Kim Kyu Hwan Oh Se‐Hee Lee 《Liver Transplantation》2016,6(15)
A new concept of pairing an active material and a mixed conductor is explored as a solid‐state battery electrode. By imbedding nano‐FeS2 domains into an amorphous LiTiS2 matrix, a hybrid power‐energy system is achieved while additionally improving upon many common solid electrode design flaws. High‐resolution transmission electron microscopy is used to probe the active material/mixed conductor interface over the course of cycling. Arguably the most beneficial development is enhancement of charge transfer, manifesting in a significantly increased exchange current as captured in a Tafel analysis. By developing a solution to active material isolation and creating a more homogenous electrode design, cycling at a high rate of C/2 for 500 cycles is obtained. Additionally, the electrode can recover full capacity simply by reducing system rate. Capacity recovery implicates a lack of active material isolation, a common problem in solid‐state batteries. 相似文献
190.
We recently showed that inter-keratin disulfide bonding plays an important role in the assembly, organization, and dynamics of keratin intermediate filaments in skin keratinocytes. In particular, cysteine 367 located in the central α-helical rod domain of keratin 14 is necessary for the formation of a stable perinuclear network of keratin filaments (with type II partner keratin 5) in skin keratinocytes analyzed by static and live cell imaging. Here, we show that two additional cysteine residues located in the non-helical head domain of K14, Cys-4 and Cys-40, also participate in inter-keratin disulfide bonding and tandemly play a key role complementary to that of Cys-367 in the assembly, organization, and dynamics of keratin filaments in skin keratinocytes in primary culture. Analysis of K14 variants with single or multiple substitutions of cysteine residues points to a spatial and temporal hierarchy in how Cys-4/Cys-40 and Cys-367 regulate keratin assembly in vitro and filament dynamics in live keratinocytes in culture. Our findings substantiate the importance and complexity of a novel determinant, namely inter-keratin disulfide bonding, for the regulation of several aspects of keratin filaments in surface epithelia. 相似文献