CDC42 Gtpase Activation Affects Hela Cell DNA Repair and Proliferation Following UV Radiation‐Induced Genotoxic Stress

Ascer, Liv Goldstein; Magalhães, Yuli Thamires; Espinha, Gisele T. da S; Osaki, Juliana Harumi; Souza, Renan Cantalice de; Forti, Fábio Luís

doi:10.1002/jcb.25166

Cited by 14 publications

(26 citation statements)

References 46 publications

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“…Our results demonstrated that ROS production increased in ADMSCs from aged rats and that this was attenuated by CASIN. These data, together with the observation that Cdc42 regulates NADPH oxidase activation (Ascer et al 2015;Qian et al 2005;Wang et al 2012b) suggests the possibility that inhibition of Cdc42 leads to an improvement of ADMSC proliferation and differentiation via suppression of NADPH oxidase activity.…”

Section: Discussionmentioning

confidence: 76%

Elevated levels of the small GTPase Cdc42 induces senescence in male rat mesenchymal stem cells

et al. 2018

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Mesenchymal stem cells (MSCs) represent a promising cell source for cellular therapy and tissue engineering and are currently being tested in a number of clinical trials for various diseases. However, like other somatic cells, MSCs age, and this senescence is accompanied by a progressive decline in stem cell function. Several lines of evidence suggest a role for the Rho family GTPase Cdc42 activity in cellular senescence processes. In the present study, we have examined aging-associated Cdc42 activity in rat adipose-derived mesenchymal stem cells (ADMSCs) and the consequences of pharmacological inhibition of Cdc42 in ADMSCs from aged rats. We demonstrate that ADMSCs show a decreased rate of cell growth and a decreased ability to differentiate into chrodrogenic, osteogenic and adipogenic cell lineages as a function of rat age. This is accompanied with an increased staining for SA-β-Gal activity and increased levels of Cdc42 bound to GTP. Treatment of ADMSCs from 24-month old rats with three Cdc42 inhibitors significantly increased proliferation rates, decreased SA-β-Gal staining, and reduced Cdc42-GTP. The Cdc42 inhibitor CASIN increased adipogenic and osteogenic differentiation potential in ADMSCs from 24-month old rats, and decreased the levels of radical oxygen species (ROS), p16 levels, F-actin, and the activity of the ERK1/2 and JNK signaling pathways that were all elevated in these cells. These data suggest that ADMSCs show increased rates of senescence as rats age that appear to be due to elevated Cdc42 activity. Thus, Cdc42 plays important roles in MSC senescence and differentiation potential, and pharmacological reduction of Cdc42 activity can, at least partially, rejuvenate aged MSCs.

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Section: Discussionmentioning

confidence: 76%

Elevated levels of the small GTPase Cdc42 induces senescence in male rat mesenchymal stem cells

et al. 2018

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“…DNA damage can be evaluated by comet assay 18 19 , which was employed in this study to determine whether HBx could result in DNA damage. As shown in Fig.…”

Section: Resultsmentioning

confidence: 99%

Hepatitis B virus X protein (HBx)-induced abnormalities of nucleic acid metabolism revealed by 1H-NMR-based metabonomics

Yue

Zhang

Cheng

et al. 2016

Sci Rep

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Hepatitis B virus X protein (HBx) plays an important role in HBV-related hepatocarcinogenesis; however, mechanisms underlying HBx-mediated carcinogenesis remain unclear. In this study, an NMR-based metabolomics approach was applied to systematically investigate the effects of HBx on cell metabolism. EdU incorporation assay was conducted to examine the effects of HBx on DNA synthesis, an important feature of nucleic acid metabolism. The results revealed that HBx disrupted metabolism of glucose, lipids, and amino acids, especially nucleic acids. To understand the potential mechanism of HBx-induced abnormalities of nucleic acid metabolism, gene expression profiles of HepG2 cells expressing HBx were investigated. The results showed that 29 genes involved in DNA damage and DNA repair were differentially expressed in HBx-expressing HepG2 cells. HBx-induced DNA damage was further demonstrated by karyotyping, comet assay, Western blotting, immunofluorescence and immunohistochemistry analyses. Many studies have previously reported that DNA damage can induce abnormalities of nucleic acid metabolism. Thus, our results implied that HBx initially induces DNA damage, and then disrupts nucleic acid metabolism, which in turn blocks DNA repair and induces the occurrence of hepatocellular carcinoma (HCC). These findings further contribute to our understanding of the occurrence of HCC.

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“…In order to certify that the selected inducers would actually activate RhoGTPases, we conducted an activity assay, to measure GTP-bound Cdc42, Rac and RhoA proteins. The following protocol was adapted from Espinha et al ., 2016 and Ascer et al ., 2015 33 , 34 . E. coli (strain BL21-DE3) bacteria were transformed via thermal shock with plasmids containing the sequence encoding the fusion protein RBD-GST (Rhotekin-Binding Domain-fused to Glutathione S-Transferase) or PBD-GST (PAK1-Binding Domain fused to Glutathione-S Transferase), which was kindly donated by Gary M. Bokoch from Scripps Research Institute, La Jolla, CA, USA.…”

Section: Methodsmentioning

confidence: 99%

“… 33 and Ascer et al . 34 . SHEDs protein lysates from a control sample were obtained from 10-mm dishes at approximately 60% confluence that were serum starved for 18 h and then treated 100 ng/ml of epidermal growth factor – EGF (Peprotech, NJ, USA) to activate Cdc42 21 ( http://www.cytoskeleton.com/cn02 ), 25 ng/ml of EGF to activate Rac1/2/3 22 ( http://www.cytoskeleton.com/cn02 ) and 30 ug/ml of calpeptin (Tocris, UK) to activate RhoA 23 ( http://www.cytoskeleton.com/cn01 ).…”

Section: Methodsmentioning

confidence: 99%

Actin cytoskeleton dynamics in stem cells from autistic individuals

Griesi-Oliveira

Suzuki

Alves

et al. 2018

Sci Rep

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Several lines of indirect evidence, such as mutations or dysregulated expression of genes related to cytoskeleton, have suggested that cytoskeletal dynamics, a process essential for axons and dendrites development, is compromised in autism spectrum disorders (ASD). However, no study has yet examined whether cytoskeleton dynamics is functionally altered in cells from ASD patients. Here we investigated the regulation of actin cytoskeleton dynamics in stem cells from human exfoliated deciduous teeth (SHEDs) of 13 ASD patients and 8 control individuals by inducing actin filament depolymerization and then measuing their reconstruction upon activation of the RhoGTPases Rac, Cdc42 or RhoA. We observed that stem cells from seven ASD individuals (53%) presented altered dymanics of filament reconstruction, including a patient recently studied by our group whose iPSC-derived neuronal cells show shorten and less arborized neurites. We also report potentially pathogenic genetic variants that might be related to the alterations in actin repolymerization dynamics observed in some patient-derived cells. Our results suggest that, at least for a subgroup of ASD patients, the dynamics of actin polymerization is impaired, which might be ultimately leading to neuronal abnormalities.

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CDC42 Gtpase Activation Affects Hela Cell DNA Repair and Proliferation Following UV Radiation‐Induced Genotoxic Stress

Cited by 14 publications

References 46 publications

Elevated levels of the small GTPase Cdc42 induces senescence in male rat mesenchymal stem cells

Elevated levels of the small GTPase Cdc42 induces senescence in male rat mesenchymal stem cells

Hepatitis B virus X protein (HBx)-induced abnormalities of nucleic acid metabolism revealed by 1H-NMR-based metabonomics

Actin cytoskeleton dynamics in stem cells from autistic individuals

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