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Various Isoforms (various + isoform)
Selected AbstractsAlternative splicing of MDM2 mRNA in lung carcinomas and lung cell linesENVIRONMENTAL AND MOLECULAR MUTAGENESIS, Issue 1 2005Mao-Wen Weng Abstract The MDM2 gene is overexpressed in several human tumors and its product may be processed into various isoforms. Recently, alternative splicing forms of MDM2 mRNA have been detected in various types of tumors. In this study, lung tissue from human non small cell lung cancers was examined for MDM2 mRNA splicing variants by nested RT-PCR. Of the 117 lung cancer tissue samples analyzed, a total of 31 (26.5%) had splice variants for the MDM2 gene, while 59 (50.4%) had undetectable levels of MDM2 transcript. Further analysis indicated that the predominant variant for 26 of the 31 samples with alternative MDM2 splicing products was MDM2-657, a splice variant lacking exons 3,11. Significant associations were found between the frequency of alternative splicing and the gender and smoking habits of the patients. Approximately 36% of male patients had alternative splicing of MDM2 compared with only 9.5% of female patients (P = 0.008); 44.2% of the smoker patients had alternative MDM2 splice forms versus 16.2% of nonsmokers (P = 0.003). Furthermore, most normal lung cell lines examined possessed only full-length MDM2 mRNA, while among several lung cancer cell lines, only H1355 and CaLu-1 cells lacked alternatively spliced MDM2 transcripts. When H1355 cells were treated in vitro with the cigarette smoke carcinogen benzo[a]pyrene (B[a]P) or the B[a]P metabolite benzo[a]pyrene diolepoxide (BPDE), three MDM2 splicing products were detected by nested RT-PCR. Finally, with the use of several specific inhibitors, we found that BPDE-induced MDM2 mRNA alternative splicing in H1355 cells may occur through the PI3K or MAPK pathway. Overall, our results suggest that carcinogens present in cigarette smoke increase the risk of alternative MDM2 splicing, which is highly associated with lung cancer. Environ. Mol. Mutagen., 2005. © 2005 Wiley-Liss, Inc. [source] Assessment of Growth, Physiological and Biochemical Parameters and Activities of Antioxidative Enzymes in Salinity Tolerant and Sensitive Basmati Rice VarietiesJOURNAL OF AGRONOMY AND CROP SCIENCE, Issue 6 2007M. P. Singh Abstract This investigation was undertaken to compare the level of salinity tolerance of the newly bred CSR-30 basmati rice variety with that of the salinity sensitive HBC-19 and Pokkali rice varieties. Twenty-one-day-old hydroponically raised seedlings at 6 and 12 dS m,1 were investigated for growth, photosynthetic rate, chlorophyll content, ribulose-1,5-bisphosphate carboxylase-oxygenase (Rubisco) activity, relative water content (RWC), membrane stability index (MSI), lipid peroxidation, Na/K ratio and activities and gene expression of various isoforms of antioxidative enzymes. Salinity stress led to reduction in shoot length, leaf area, dry weight, RWC, MSI, rate of photosynthesis, chlorophyll content and Rubisco activity in all the three rice varieties. The levels of reduction in these parameters were maximal in HBC-19 followed by those in CSR-30 and Pokkali respectively. Cumulative superoxide dismutase (SOD) activity increased in Pokkali and CSR-30 in consonance with increase in salinity stress while it decreased in HBC-19. The Mn-SOD activity however, was enhanced in all three varieties in the presence of salinity stress while the activities of Fe-SOD, Cu/Zn-SOD and ascorbate peroxidase were decreased in HBC-19 when compared with CSR-30 and Pokkali. The activity of catalase (CAT) was higher in HBC-19 when compared with its activity in CSR-30 and Pokkali. The levels of gene expressions of the three isoforms of SOD ascertained by reverse transcriptase polymerase chain reaction were not necessarily indicative of the activities of the corresponding enzymes. Thus, despite the maximal enhancement in gene expression of Fe-SOD in HBC-19 in response to salinity stress, the activity of this enzyme in HBC-19 remained low. Similarly, despite a marginal increase in gene expression of Cu-Zn SOD in the three varieties, its activity was significantly higher in Pokkali and CSR-30 when compared with that in HBC-19. A significant enhancement in the activity of CAT at 12 dS m,1 in HBC-19 when compared with CSR-30 and Pokkali might confer a degree of tolerance to H2O2 stress in this variety in the presence of higher levels of NaCl at the seedling stage. [source] Novel targets for valproic acid: up-regulation of melatonin receptors and neurotrophic factors in C6 glioma cellsJOURNAL OF NEUROCHEMISTRY, Issue 5 2005Lyda M. Rincón Castro Abstract Valproic acid (VPA) is a potent anti-epileptic and effective mood stabilizer. It is known that VPA enhances central GABAergic activity and activates the mitogen-activated protein kinase,extracellular signal-regulated kinase (MAPK,ERK) pathway. It can also inhibit various isoforms of the enzyme, histone deacetylase (HDAC), which is associated with modulation of gene transcription. Recent in vivo studies indicate a neuroprotective role for VPA, which has been found to up-regulate the expression of brain-derived neurotrophic factor (BDNF) in the rat brain. Given the interaction between the pineal hormone, melatonin, and GABAergic systems in the central nervous system, the effects of VPA on the expression of the mammalian melatonin receptor subtypes, MT1 and MT2, were examined in rat C6 glioma cells. The effects of VPA on the expression of glial cell line-derived neurotrophic factor (GDNF) and BDNF were also examined. RT-PCR studies revealed a significant induction of melatonin MT1 receptor mRNA in C6 cells following treatment with 3 or 5 mm VPA for 24 h or 5 mm VPA for 48 h. Western analysis and immunocytochemical detection confirmed that the VPA-induced increase in MT1 mRNA results in up-regulation of MT1 protein expression. Blockade of the MAPK,ERK pathway by PD98059 enhanced the effect of VPA on MT1 expression, suggesting a negative role for this pathway in MT1 receptor regulation. In addition, significant increases in BDNF, GDNF and HDAC mRNA expression were observed after treatment with VPA for 24 or 48 h. Taken together, the present findings suggest that the neuroprotective properties of VPA involve modulation of neurotrophic factors and receptors for melatonin, which is also thought to play a role in neuroprotection. Moreover, the foregoing suggests that combinations of VPA and melatonin could provide novel therapeutic strategies in neurological and psychiatric disorders. [source] Interaction of Drugs and Chinese Herbs: Pharmacokinetic Changes of Tolbutamide and Diazepam Caused by Extract of Angelica dahuricaJOURNAL OF PHARMACY AND PHARMACOLOGY: AN INTERNATI ONAL JOURNAL OF PHARMACEUTICAL SCIENCE, Issue 8 2000KAZUHISA ISHIHARA The inhibitory effects of Angelica dahurica root extract on rat liver microsomal cytochrome P450 and drug-drug interactions were studied. The 2,- and 16,-hydroxylase activity of testosterone were most strongly inhibited, with 17.2% and 28.5% of their activity remaining, respectively, after oral administration of A. dahurica extract at a 1 g kg,1 dose. 6,-Hydroxylase activity was also inhibited, with 70% of its activity remaining, under the same conditions. In addition, treatment with the extract inhibited the metabolism of tolbutamide, nifedipine and bufuralol. These results showed that the extract inhibited the various isoforms of cytochrome P450 such as CYP2C, CYP3A and CYP2D1. The A. dahurica extract delayed elimination of tolbutamide after intravenous administration at a 10 mg kg,1 dose to rats. Thus, the extract altered the liver intrinsic clearance. It had little effect, however, on the pharmacokinetic parameters of diazepam after intravenous administration at 10 mg kg,1. Since diazepam showed high clearance, it underwent hepatic blood flow rate-limited metabolism. Therefore, the change of intrinsic clearance had little effect on hepatic clearance. However, the Cmax value after oral administration of diazepam with extract treatment was four times that with non-treatment. It was suggested that the first-pass effect was changed markedly by the extract. High-dose (1 g kg,1), but not low dose (0.3 g kg,1), administration of A. dahurica extract increased significantly the duration of rotarod disruption following intravenous administration of diazepam at 5 mg kg,1. It was concluded that administration of A. dahurica extract has the potential to interfere with the metabolism, by liver cytochrome P450, of other drugs. [source] Structural Features of the NAD-Dependent In Situ Retinoic Acid Supply System in Esophageal MucosaALCOHOLISM, Issue 2010Hirokazu Yokoyama Background:, We previously reported that an NAD-dependent in situ retinoic acid supply system, which comprises some isoforms of alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) and provides retinoic acid from retinol via a 2-step oxidation process, exists in the rat esophagus. Herein, their isoforms responsible for the pathway and its localization in the rat esophagus was examined. Methods:, The expressions of mRNAs of various isoforms of ADH and ALDH were examined in the fraction mainly comprising mucosal layer of the rat esophagus by RT-PCR. Expression levels of Class IV ADH and ALDH 1A1 were compared between the fractions and that mainly comprising muscle layer of the rat esophagus by quantitative PCR. The catalytic activities producing retinoic acid from retinal were compared between the 2 fractions and its optimum pH was also determined. Results:, Classes I, III, and IV ADHs and ALDHs 1A1 and 3A1 were predominant isoforms in the rat esophageal mucosa. The expression levels of mRNA of Class IV ADH and ALDH 3A1 were significantly higher in the mucosal than in the muscle layer. Consistently, the catalytic activities producing retinoic acid from retinal were significantly higher in the former than the latter. The optimum pH of the process was 9.0. Conclusions:, Considering the affinities for retinol and retinal of ADHs and ALDHs expressed in the rat esophagus, the NAD-dependent in situ retinoic acid supply system in the rat esophagus is thought to comprise Class IV ADH and ALDH 1A1. In the rat esophagus, the system exists predominantly in the mucosal layer. [source] The transcription factor CREM, and cAMP regulate promoter activity of the Na,K-ATPase ,4 isoformMOLECULAR REPRODUCTION & DEVELOPMENT, Issue 11 2006Marianna Rodova Abstract The Na,K-ATPase is an essential enzyme of the plasma membrane that plays a key role in numerous cell processes that depend on the transcellular gradients of Na+ and K+. Among the various isoforms of the catalytic subunit of the Na,K-ATPase, ,4 exhibits the most limited pattern of expression, being restricted to male germ cells. Activity of ,4 is essential for sperm function, and ,4 is upregulated during spermatogenesis. The present study addressed the transcriptional control of the human Na,K-ATPase ,4 gene, ATP1A4. We describe that a 5, untranslated region of the ATP1A4 gene (designated ,339/+480 based on the ATP1A4 transcription initiation site) has promoter activity in luciferase reporter assays. Computer analysis of this promoter region revealed consensus sites (CRE) for the cyclic AMP (cAMP) response element modulator (CREM). Accordingly, dibutyryl cAMP (db-cAMP) and ectopic expression of CREM,, a testis specific splice variant of CREM were able to activate the ATP1A4 promoter driven expression of luciferase in HEK 293 T, JEG-3 and GC-1 cells. Further characterization of the effect of db-cAMP and CREM, on deleted constructs of the ATP1A4 promoter (,339/+80, and +25/+480), and on the ,339/+480 region carrying mutations in the CRE sites showed that db-cAMP and CREM, effect required the CRE motif located 263 bp upstream the transcription initiation site. EMSA experiments confirmed the CRE sequence as a bonafide CREM, binding site. These results constitute the first demonstration of the transcriptional control of ATP1A4 gene expression by cAMP and by CREM,, a transcription factor essential for male germ cell gene expression. Mol. Reprod. Dev. 73: 1435,1447, 2006. © 2006 Wiley-Liss, Inc. [source] Non-solid oncogenes in solid tumors: EML4,ALK fusion genes in lung cancerCANCER SCIENCE, Issue 12 2008Hiroyuki Mano It is generally accepted that recurrent chromosome translocations play a major role in the molecular pathogenesis of hematological malignancies but not of solid tumors. However, chromosome translocations involving the e26 transformation-specific sequence transcription factor loci have been demonstrated recently in many prostate cancer cases. Furthermore, through a functional screening with retroviral cDNA expression libraries, we have discovered the fusion-type protein tyrosine kinase echinoderm microtubule-associated protein like-4 (EML4),anaplastic lymphoma kinase (ALK) in non-small cell lung cancer (NSCLC) specimens. A recurrent chromosome translocation, inv(2)(p21p23), in NSCLC generates fused mRNA encoding the amino-terminal half of EML4 ligated to the intracellular region of the receptor-type protein tyrosine kinase ALK. EML4,ALK oligomerizes constitutively in cells through the coiled coil domain within the EML4 region, and becomes activated to exert a marked oncogenicity both in vitro and in vivo. Break and fusion points within the EML4 locus may diverge in NSCLC cells to generate various isoforms of EML4,ALK, which may constitute ~5% of NSCLC cases, at least in the Asian ethnic group. In the present review I summarize how detection of EML4,ALK cDNA may become a sensitive diagnostic means for NSCLC cases that are positive for the fusion gene, and discuss whether suppression of ALK enzymatic activity could be an effective treatment strategy against this intractable disorder. (Cancer Sci 2008; 99: 2349,2355) [source] Snake venom hyaluronidase: a therapeutic targetCELL BIOCHEMISTRY AND FUNCTION, Issue 1 2006K. Kemparaju Abstract The diffusion of toxins from the site of a bite into the circulation is essential for successful envenomation. Degradation of hyaluronic acid in the extracellular matrix (ECM) by venom hyaluronidase is a key factor in this diffusion. Hyaluronidase not only increases the potency of other toxins but also damages the local tissue. In spite of its important role, little attention has been paid to this enzyme. Hyaluronidase exists in various isoforms and generates a wide range of hyaluronic acid degradation products. This suggests that beyond its role as a spreading factor venom hyaluronidase deserves to be explored as a possible therapeutic target for inhibiting the systemic distribution of venom and also for minimizing local tissue destruction at the site of the bite. Copyright © 2005 John Wiley & Sons, Ltd. [source] | |||||||||||||