scholarly journals A novel member of the SAF (scaffold attachment factor)-box protein family inhibits gene expression and induces apoptosis

2007 ◽  
Vol 407 (3) ◽  
pp. 355-362 ◽  
Author(s):  
Ching Wan Chan ◽  
Youn-Bok Lee ◽  
James Uney ◽  
Andrea Flynn ◽  
Jonathan H. Tobias ◽  
...  
Keyword(s):  
Gene Expression ◽  
Rna Binding ◽  
Chromatin Condensation ◽  
Inhibitory Effect ◽  
Binding Motif ◽  
Inhibitory Effects ◽  
Cytochrome C Release ◽  
Interacting Protein ◽  
Induced Apoptosis ◽  
Attachment Factor

The SLTM [SAF (scaffold attachment factor)-like transcription modulator] protein contains a SAF-box DNA-binding motif and an RNA-binding domain, and shares an overall identity of 34% with SAFB1 {scaffold attachment factor-B1; also known as SAF-B (scaffold attachment factor B), HET [heat-shock protein 27 ERE (oestrogen response element) and TATA-box-binding protein] or HAP (heterogeneous nuclear ribonucleoprotein A1-interacting protein)}. Here, we show that SLTM is localized to the cell nucleus, but excluded from nucleoli, and to a large extent it co-localizes with SAFB1. In the nucleus, SLTM has a punctate distribution and it does not co-localize with SR (serine/arginine) proteins. Overexpression of SAFB1 has been shown to exert a number of inhibitory effects, including suppression of oestrogen signalling. Although SLTM also suppressed the ability of oestrogen to activate a reporter gene in MCF-7 breast-cancer cells, inhibition of a constitutively active β-galactosidase gene suggested that this was primarily the consequence of a generalized inhibitory effect on transcription. Measurement of RNA synthesis, which showed a particularly marked inhibition of [3H]uridine incorporation into mRNA, supported this conclusion. In addition, analysis of cell-cycle parameters, chromatin condensation and cytochrome c release showed that SLTM induced apoptosis in a range of cultured cell lines. Thus the inhibitory effects of SLTM on gene expression appear to result from generalized down-regulation of mRNA synthesis and initiation of apoptosis consequent upon overexpressing the protein. While indicating a crucial role for SLTM in cellular function, these results also emphasize the need for caution when interpreting phenotypic changes associated with manipulation of protein expression levels.

scholarly journals The Study of Anticancer Activity of Satureja Khuzestanica Alcoholic Extracts On Expression of Bcl2 And Bax Genes In The Pc3 Cell Line

2021 ◽  
Author(s):  
Saman Kazemi ◽  
asghar tanomand ◽  
Hossein Soltanzadeh ◽  
Gholamreza Shahsavari
Keyword(s):  
Gene Expression ◽  
Prostate Cancer ◽  
Cancer Cells ◽  
Rna Extraction ◽  
Inhibitory Effect ◽  
Chloroform Extract ◽  
Antioxidant Compounds ◽  
Anticancer Properties ◽  
Satureja Khuzestanica ◽  
Induced Apoptosis

Abstract Introduction: Prostate cancer is the most common cancer among men after lung cancer. It has grown in Iran in recent years. The use of medicinal plants is one of the most useful ways that causes the least side effects. Due to high levels of antioxidant compounds, Satureja khuzestanica is a good source for drug use to treat and prevent the development and progress of cancers. The aim of the present study was to evaluate the anti-cancer property of Satureja khuzestanica extract on the expression of Bcl2 and Bax genes in prostate cancer cell lines.Methodology: After collecting the plant in spring, the chloroform extract was prepared by rotary device. PC3 cancer cells were incubated at different concentrations of the extract for 24 hours. The inhibitory effect of the extract was evaluated using MTT assay as IC50. To evaluate apoptosis, the level of expression of Bax and BCL-2 genes after RNA extraction and transformation to cDNA were evaluated using Real Time PCR. All data were analyzed using REST software.Results: The results revealed a direct and significant relationship between the two variables of drug composition and rate of PC3 cell death. This composition increased Bax gene expression and decreased BCL-2 gene expression and induced apoptosis (P <0.05).Discussion and Conclusion: Based on the results, Satureja khuzestanica extract is likely to have anticancer properties and seems to be a new drug for killing prostate cancer cells.

scholarly journals Hypericin Enhances Paclitaxel-Induced B16-F10 Cell Apoptosis by Activating a Cytochrome c Release–Dependent Pathway

2021 ◽  
Vol 12 ◽  
Author(s):  
Liyun Sun ◽  
Zixuan Li ◽  
Huoli Shang ◽  
Xiujuan Xin
Keyword(s):  
Cytochrome C ◽  
Caspase 3 ◽  
Chromatin Condensation ◽  
Combined Treatment ◽  
Inhibitory Effect ◽  
Combination Group ◽  
Oxygen Species ◽  
Apoptotic Body ◽  
Cytochrome C Release ◽  
Dependent Pathway

The enhanced inhibitory effect of paclitaxel (PTX) combined with hypericin (HY) on B16-F10 cells may be realized through the ROS-related cytochrome c release pathway. The apoptotic characteristics of the B16-F10 cells, such as DNA fragmentation, chromatin condensation, and apoptotic body formation, were all enhanced in the combined treatment group. Further investigation showed that the combination of paclitaxel and HY could increase the level of mitochondrial damage and the concentration of cytochrome c, causing the expression of caspase-3 and the cleavage of PARP.1. Compared with paclitaxel or HY alone, the level of reactive oxygen species (ROS) increased significantly, while glutathione reductase (GR) activity and intracellular glutathione (GSH) levels decreased significantly in the combination group.

scholarly journals Astragaloside IV inhibits palmitic acid-induced apoptosis through regulation of calcium homeostasis in mice podocytes

2021 ◽  
Author(s):  
Yingjun Zang ◽  
Shuang Liu ◽  
Aili Cao ◽  
Xiangyu Shan ◽  
Wenjuan Deng ◽  
...  
Keyword(s):  
Palmitic Acid ◽  
Mitochondrial Membrane ◽  
Protective Efficacy ◽  
Calcium Ionophore ◽  
Cytosolic Calcium ◽  
Inhibitory Effect ◽  
Astragaloside Iv ◽  
Cytochrome C Release ◽  
Induced Apoptosis ◽  
Cellular Ca

AbstractLoss of podocytes is a hallmark of diabetic nephropathy, and a growing body of evidence indicates that podocytes are susceptible to palmitic acid (PA). We have previously shown that AS-IV inhibited PA-induced podocyte apoptosis by activating sarcoendoplasmic reticulum Ca2+ ATPase (SERCA), which indicate calcium regulation may involve in the process. Immunofluorescence staining, Western blot and flow cytometry were used to measure the protective efficacy of AS-IV to ameliorate PA-induced ER stress and podocyte apoptosis. Meanwhile, AS-IV inhibited cytochrome c release, decreased mitochondrial membrane potential, accompany with the depletion of endoplasmic reticulum Ca2+ and elevation of cytosolic and mitochondrial Ca2+. Sequestration of cytosolic calcium with BAPTA-AM limited the response of podocyte apoptosis, while during the process the effect of AS-IV was also restrained. In contrast, elevation of cytosolic calcium with calcium ionophore ionomycin was depressed by AS-IV addition. Furthermore, inhibiting TRPC6 expression with SKF96365 or TRPC6 siRNA counteracted the beneficial effect of AS-IV. Our study provides further evidence to conclude the inhibitory effect of AS-IV to podocyte apoptosis is Ca2+-dependent. And the efficacy correlates with inhibiting TRPC6-mediated Ca2+ influx, and then cellular Ca2+ disturbance was coordinated.

Heat-shock protein 70 attenuates nitric oxide-induced apoptosis in RAW macrophages by preventing cytochrome c release

2002 ◽  
Vol 362 (3) ◽  
pp. 635-641 ◽  
Author(s):  
Sabine D. KLEIN ◽  
Bernhard BRÜNE
Keyword(s):  
Nitric Oxide ◽  
Cell Cycle ◽  
Heat Shock ◽  
Cytochrome C ◽  
Heat Shock Protein ◽  
Cytochrome C Release ◽  
Interacting Protein ◽  
P53 Expression ◽  
Induced Apoptosis ◽  
Hoechst Staining

Heat-shock protein (Hsp) 70 is an inhibitor of apoptosis and has been shown to protect against nitric oxide-mediated toxicity. To gain mechanistic insights into the actions of Hsp70, we stably transfected RAW 264.7 mouse macrophages with the human Hsp70 gene and investigated critical steps in the progression towards cell demise. Incubation of control and Hsp70-transfected macrophages with S-nitrosoglutathione induced accumulation of the tumour suppressor p53, expression of p21WAF1/CIP1 (where WAF1 corresponds to wild-type p53-activated fragment 1 and CIP1 corresponds to cyclin-dependent kinase-interacting protein 1) and G1 cell-cycle arrest. However, cytochrome c translocation to the cytosol and activation of caspase 9 and caspase 3 were markedly reduced in Hsp70-overexpressing cells. In addition, changes in nuclear morphology, as determined by Hoechst staining, and the appearance of cells in the sub-G1 phase were diminished in Hsp70-overexpressing cells compared with controls. We conclude that, in macrophages, Hsp70 interferes with cytochrome c release from mitochondria and, thereby, prevents nitric oxide-induced apoptosis, but leaves p53 accumulation and interference in the cell cycle intact.

scholarly journals The RNA-binding Protein HuR Stabilizes Cytosolic Phospholipase A2α mRNA under Interleukin-1β Treatment in Non-small Cell Lung Cancer A549 Cells

2011 ◽  
Vol 286 (41) ◽  
pp. 35499-35508 ◽  
Author(s):  
Wan-Lin Liao ◽  
Wei-Chiao Wang ◽  
Wen-Chang Chang ◽  
Joseph T. Tseng
Keyword(s):  
Gene Expression ◽  
P38 Mapk ◽  
Rna Binding ◽  
A549 Cells ◽  
Inhibitory Effect ◽  
Mapk Signaling ◽  
Small Cell Lung ◽  
Target Mrnas ◽  
Cytosolic Phospholipase ◽  
Mrna And Protein Expression

The activation of cytosolic phospholipase A2α (cPLA2α) plays an important role in initiating the inflammatory response. The regulation of cPLA2α mRNA turnover has been proposed to control cPLA2α gene expression under cytokine and growth factor stimulation. However, the detailed mechanism is still unknown. In this report, we have demonstrated that the cPLA2α mRNA stability was increased under IL-1β treatment in A549 cells. By using EMSAs, HuR was identified as binding with the cPLA2α mRNA 3′-UTR, and the binding region was located at nucleotides 2716–2807, a fragment containing AUUUA flanked by U-rich sequences. IL-1β treatment enhanced the association of cPLA2α mRNA with cytosolic HuR. The reduction of HuR expression by RNA interference technology inhibited IL-1β-induced cPLA2α mRNA and protein expression. Furthermore, blocking the p38 MAPK signaling pathway with SB203580 abolished the effect of IL-1β-induced cPLA2α gene expression. Phosphorylation at residue Thr-118 of HuR is crucial in regulating the interaction between HuR and its target mRNAs. Mutation of HuR Thr-118 reduced the association between HuR and cPLA2α mRNA under IL-1β treatment. This inhibitory effect was also observed in binding with COX-2 mRNA. This result indicated that p38 MAPK-mediated Thr-118 phosphorylation may play a key role in regulating the interaction of HuR with its target mRNAs in inflammation.

Insulin inhibits glucocorticoid-stimulated L-type 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase gene expression by activation of the c-Jun N-terminal kinase pathway

2001 ◽  
Vol 353 (2) ◽  
pp. 267-273 ◽  
Author(s):  
Elisabet DE LOS PINOS ◽  
Silvia FERNÁNDEZ DE MATTOS ◽  
Manel JOAQUIN ◽  
Albert TAULER
Keyword(s):  
Gene Expression ◽  
Protein Kinase ◽  
Glucocorticoid Receptor ◽  
Map Kinase ◽  
Inhibitory Effect ◽  
Interacting Protein ◽  
Extracellular Signal ◽  
Mitogen Activated Protein ◽  
Ribosomal S6 Kinase ◽  
Phosphoinositide 3 Kinase

The hepatic isoform of 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase (PF2K/Fru-2,6-BPase) is transcriptionally stimulated by glucocorticoids, whereas insulin blocks this stimulatory effect. Although this inhibitory effect has been extensively reported, nothing is known about the signalling pathway responsible. We have used well-characterized inhibitors for proteins involved in different signalling cascades to assess the involvement of these pathways on the transcriptional regulation of glucocorticoid-stimulated PF2K/Fru-2,6-BPase by insulin. Our results demonstrate that the phosphoinositide 3-kinase, p70/p85 ribosomal S6 kinase, extracellular signal-regulated protein kinase (ERK)1/2 and p38 mitogen-activated protein (MAP) kinase pathways are not involved in the inhibitory effect of insulin on glucocorticoid-stimulated PF2K/Fru-2,6-BPase. To evaluate the implication of the MAP kinase/ERK kinase (MEK)-4Őstress-activated protein kinaseŐc-Jun-N-terminal protein kinase (‘JNKŐSAPK’) pathway we overexpressed the N-terminal JNK-binding domain of the JNK-interacting protein 1 (‘JIP-1’), demonstrating that activation of JNK is necessary for the insulin inhibitory effect. Moreover, overexpression of MEK kinase 1 and JNKŐhaemagglutinin resulted in the inhibition of the glucocorticoid-stimulated PF2K/Fru-2,6-BPase. These results provide clear and specific evidence for the role of JNK in the insulin inhibition of glucocorticoid-stimulated PF2K/Fru-2,6-BPase gene expression. In addition, we performed experiments with a mutant of the glucocorticoid receptor in which the JNK phosphorylation target Ser-246 had been mutated to Ala. Our results demonstrate that the phosphorylation of the glucocorticoid receptor on Ser-246 is not responsible for the JNK repression of glucocorticoid-stimulated PF2K/Fru-2,6-BPase gene expression.

scholarly journals Inhibitory Effect of Modified 5′-Capped Short RNA Fragments on Influenza Virus RNA Polymerase Gene Expression

2001 ◽  
Vol 12 (6) ◽  
pp. 353-358 ◽  
Author(s):  
Motoki Tado ◽  
Takayuki Abe ◽  
Toshifumi Hatta ◽  
Masahide Ishikawa ◽  
Susumu Nakada ◽  
...  
Keyword(s):  
Gene Expression ◽  
Influenza Virus ◽  
Rna Polymerase ◽  
Nuclease Activity ◽  
Inhibitory Effect ◽  
Polymerase Gene ◽  
Inhibitory Effects ◽  
Virus Rna ◽  
Rna Fragments ◽  
Rna Polymerase Gene

We have shown previously that the 5′-capped short phosphodiester RNA fragments, Cap decoy, (Gm 12 nt) are potent inhibitors of influenza virus RNA polymerase gene expression. Here we investigate the modified capped RNA derivative containing phosphorothioate oligonucleotides (Cap decoy) as a potential influenza virus RNA polymerase inhibitor. The modified 5′-capped short phosphorothioate RNA fragments (Gms 12–15 nt) with the 5′-capped structure (m7GpppGm) were synthesized by T7 RNA polymerase. The 5′-capped short RNA fragments (Gms 12–15 nt) were encapsulated in liposome particulates and tested for their inhibitory effects on influenza virus RNA polymerase gene expression in the clone 76 cells. The 12–15 nt long Gms RNA fragments showed highly inhibitory effects. By contrast, the inhibitory effects of the 13 nt long short RNA fragments (Gm 13 nt) were considerably less in comparison with the 5′-capped short phosphorothioate RNA fragments (Gms 12–15 nt). In particular, the various Gms RNA chain lengths showed no significant differences in the inhibition of influenza virus RNA polymerase gene expression. Furthermore, the capped RNA with a phosphorothioate backbone was resistant to nuclease activity. These phosphorothioate RNA fragments exhibited higher inhibitory activity than the 5′-capped short RNA fragments (Gm 12 nt). These decoys may prove to be useful in anti-influenza virus therapeutics.

The expression of the cold shock protein RNA binding motif protein 3 is transcriptionally responsive to organ temperature in mice

2020 ◽  
Vol 27 ◽  
Author(s):  
Ayako Ushio ◽  
Ko Eto
Keyword(s):  
Gene Expression ◽  
Body Temperature ◽  
Rna Binding ◽  
Mild Hypothermia ◽  
Reproductive Organs ◽  
The Body ◽  
Cold Shock Protein ◽  
Binding Motif ◽  
Liver And Kidney ◽  
The Brain

Background: Mild hypothermia, i.e. maintenance of organ temperature by up to 8°C lower than body temperature, is a critical strategy for exerting some functions of the cells and organs normally, and is an useful therapy for recovering properly from some diseases, including myocardial infarction, cardiac arrest, brain injury, and ischemic stroke. Nevertheless, there were no focusses so far on organ temperature and potential responses of gene expression to organ temperature in organs of homeothermic animals that survive under normal conditions. Objective: The present study aimed to assess organ temperature in homeothermic animals and evaluate the effect of their organ temperature on the expression of the cold shock protein RNA binding motif protein 3 (RBM3), and to gain insights into the organ temperature-mediated regulation of RBM3 gene transcription via Nuclear factor β-light-chain-enhancer of activated B cells (NF-κB) p65, which had been identified as a transcription factor that is activated by undergoing the Ser276 phosphorylation and promotes the RBM3 gene expression during mild hypothermia. Methods: We measured the temperature of several organs, where RBM3 expression was examined, in female and male mice. Next, in male mice, we tested NF-κB p65 expression and its Ser276 phosphorylation in organs that have their lower temperature than body temperature and compared them with those in organs that have their temperature near body temperature. Results: Organ temperature was around 32°C in the brain and reproductive organs, which is lower than the body temperature, and around 37°C in the heart, liver, and kidney, which is comparable to the body temperature. The expression of RBM3 was detected greatly in the brain and reproductive organs with their organ temperature of around 32°C, and poorly in the heart, liver, and kidney with their organ temperature of around 37°C. In accordance with the changes in the RBM3 expression, the NF-κB p65 Ser276 phosphorylation was detected more greatly in the testis and brain with their organ temperature of around 32°C, than in the heart, liver, and kidney with their organ temperature of around 37°C, although the NF-κB p65 expression was unchanged among all the organs tested. Discussion: Our data suggested that organ temperature lower than body temperature causes the expression of RBM3 in the brain and reproductive organs of mice, and that lower organ temperature causes the NF-κB p65 activation through the Ser276 phosphorylation, resulting in an increase in the RBM3 gene transcription, in the brain and reproductive organs of mice. Conclusion: The study may present the possibility that organ temperature-induced alterations in gene expression are organ specific in homeotherms and the possibility that organ temperature-induced alterations in gene expression are transcriptionally regulated in some organs of homeotherms.

Sign in / Sign up

Export Citation Format

Share Document