Enhanced p62-NRF2 Feedback Loop due to Impaired Autophagic Flux Contributes to Arsenic-Induced Malignant Transformation of Human Keratinocytes
Identifieur interne : 000778 ( Pmc/Checkpoint ); précédent : 000777; suivant : 000779Enhanced p62-NRF2 Feedback Loop due to Impaired Autophagic Flux Contributes to Arsenic-Induced Malignant Transformation of Human Keratinocytes
Auteurs : Xiafang Wu [République populaire de Chine] ; Ru Sun [République populaire de Chine] ; Huihui Wang [République populaire de Chine] ; Bei Yang [République populaire de Chine] ; Fang Wang [République populaire de Chine] ; Hongtao Xu [République populaire de Chine] ; Shimin Chen [République populaire de Chine] ; Rui Zhao [République populaire de Chine] ; Jingbo Pi [République populaire de Chine] ; Yuanyuan Xu [République populaire de Chine]Source :
- Oxidative Medicine and Cellular Longevity [ 1942-0900 ] ; 2019.
Abstract
Chronic exposure to arsenic induces a variety of cancers, particularly in the skin. Autophagy is a highly conserved process which plays a dual role in tumorigenesis. In the present study, we found that chronic exposure to an environmentally relevant dose of arsenite induced malignant transformation of human keratinocytes (HaCaT) with dysregulated autophagy as indicated by an increased number of autophagosomes, activation of mTORC1 pathway, and elevated protein levels of p62 and LC3II. Meanwhile, arsenite-transformed cells showed lower intracellular levels of reactive oxygen species compared with control. Silencing
Url:
DOI: 10.1155/2019/1038932
PubMed: 31781319
PubMed Central: 6875345
Affiliations:
Links toward previous steps (curation, corpus...)
Links to Exploration step
PMC:6875345Le document en format XML
<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Enhanced p62-NRF2 Feedback Loop due to Impaired Autophagic Flux Contributes to Arsenic-Induced Malignant Transformation of Human Keratinocytes</title>
<author><name sortKey="Wu, Xiafang" sort="Wu, Xiafang" uniqKey="Wu X" first="Xiafang" last="Wu">Xiafang Wu</name>
<affiliation wicri:level="1"><nlm:aff id="I1">School of Public Health, China Medical University, China</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea>School of Public Health, China Medical University</wicri:regionArea>
<wicri:noRegion>China Medical University</wicri:noRegion>
</affiliation>
<affiliation wicri:level="1"><nlm:aff id="I2">The First Hospital of China Medical University, China</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea>The First Hospital of China Medical University</wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Sun, Ru" sort="Sun, Ru" uniqKey="Sun R" first="Ru" last="Sun">Ru Sun</name>
<affiliation wicri:level="1"><nlm:aff id="I1">School of Public Health, China Medical University, China</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea>School of Public Health, China Medical University</wicri:regionArea>
<wicri:noRegion>China Medical University</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Wang, Huihui" sort="Wang, Huihui" uniqKey="Wang H" first="Huihui" last="Wang">Huihui Wang</name>
<affiliation wicri:level="1"><nlm:aff id="I1">School of Public Health, China Medical University, China</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea>School of Public Health, China Medical University</wicri:regionArea>
<wicri:noRegion>China Medical University</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Yang, Bei" sort="Yang, Bei" uniqKey="Yang B" first="Bei" last="Yang">Bei Yang</name>
<affiliation wicri:level="1"><nlm:aff id="I3">College of Basic Medical Sciences, China Medical University, China</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea>College of Basic Medical Sciences, China Medical University</wicri:regionArea>
<wicri:noRegion>China Medical University</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Wang, Fang" sort="Wang, Fang" uniqKey="Wang F" first="Fang" last="Wang">Fang Wang</name>
<affiliation wicri:level="1"><nlm:aff id="I1">School of Public Health, China Medical University, China</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea>School of Public Health, China Medical University</wicri:regionArea>
<wicri:noRegion>China Medical University</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Xu, Hongtao" sort="Xu, Hongtao" uniqKey="Xu H" first="Hongtao" last="Xu">Hongtao Xu</name>
<affiliation wicri:level="1"><nlm:aff id="I3">College of Basic Medical Sciences, China Medical University, China</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea>College of Basic Medical Sciences, China Medical University</wicri:regionArea>
<wicri:noRegion>China Medical University</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Chen, Shimin" sort="Chen, Shimin" uniqKey="Chen S" first="Shimin" last="Chen">Shimin Chen</name>
<affiliation wicri:level="1"><nlm:aff id="I1">School of Public Health, China Medical University, China</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea>School of Public Health, China Medical University</wicri:regionArea>
<wicri:noRegion>China Medical University</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Zhao, Rui" sort="Zhao, Rui" uniqKey="Zhao R" first="Rui" last="Zhao">Rui Zhao</name>
<affiliation wicri:level="1"><nlm:aff id="I4">School of Forensic Medicine, China Medical University, China</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea>School of Forensic Medicine, China Medical University</wicri:regionArea>
<wicri:noRegion>China Medical University</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Pi, Jingbo" sort="Pi, Jingbo" uniqKey="Pi J" first="Jingbo" last="Pi">Jingbo Pi</name>
<affiliation wicri:level="1"><nlm:aff id="I1">School of Public Health, China Medical University, China</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea>School of Public Health, China Medical University</wicri:regionArea>
<wicri:noRegion>China Medical University</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Xu, Yuanyuan" sort="Xu, Yuanyuan" uniqKey="Xu Y" first="Yuanyuan" last="Xu">Yuanyuan Xu</name>
<affiliation wicri:level="1"><nlm:aff id="I1">School of Public Health, China Medical University, China</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea>School of Public Health, China Medical University</wicri:regionArea>
<wicri:noRegion>China Medical University</wicri:noRegion>
</affiliation>
</author>
</titleStmt>
<publicationStmt><idno type="wicri:source">PMC</idno>
<idno type="pmid">31781319</idno>
<idno type="pmc">6875345</idno>
<idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6875345</idno>
<idno type="RBID">PMC:6875345</idno>
<idno type="doi">10.1155/2019/1038932</idno>
<date when="2019">2019</date>
<idno type="wicri:Area/Pmc/Corpus">000762</idno>
<idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">000762</idno>
<idno type="wicri:Area/Pmc/Curation">000762</idno>
<idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Curation">000762</idno>
<idno type="wicri:Area/Pmc/Checkpoint">000778</idno>
<idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Checkpoint">000778</idno>
</publicationStmt>
<sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Enhanced p62-NRF2 Feedback Loop due to Impaired Autophagic Flux Contributes to Arsenic-Induced Malignant Transformation of Human Keratinocytes</title>
<author><name sortKey="Wu, Xiafang" sort="Wu, Xiafang" uniqKey="Wu X" first="Xiafang" last="Wu">Xiafang Wu</name>
<affiliation wicri:level="1"><nlm:aff id="I1">School of Public Health, China Medical University, China</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea>School of Public Health, China Medical University</wicri:regionArea>
<wicri:noRegion>China Medical University</wicri:noRegion>
</affiliation>
<affiliation wicri:level="1"><nlm:aff id="I2">The First Hospital of China Medical University, China</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea>The First Hospital of China Medical University</wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Sun, Ru" sort="Sun, Ru" uniqKey="Sun R" first="Ru" last="Sun">Ru Sun</name>
<affiliation wicri:level="1"><nlm:aff id="I1">School of Public Health, China Medical University, China</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea>School of Public Health, China Medical University</wicri:regionArea>
<wicri:noRegion>China Medical University</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Wang, Huihui" sort="Wang, Huihui" uniqKey="Wang H" first="Huihui" last="Wang">Huihui Wang</name>
<affiliation wicri:level="1"><nlm:aff id="I1">School of Public Health, China Medical University, China</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea>School of Public Health, China Medical University</wicri:regionArea>
<wicri:noRegion>China Medical University</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Yang, Bei" sort="Yang, Bei" uniqKey="Yang B" first="Bei" last="Yang">Bei Yang</name>
<affiliation wicri:level="1"><nlm:aff id="I3">College of Basic Medical Sciences, China Medical University, China</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea>College of Basic Medical Sciences, China Medical University</wicri:regionArea>
<wicri:noRegion>China Medical University</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Wang, Fang" sort="Wang, Fang" uniqKey="Wang F" first="Fang" last="Wang">Fang Wang</name>
<affiliation wicri:level="1"><nlm:aff id="I1">School of Public Health, China Medical University, China</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea>School of Public Health, China Medical University</wicri:regionArea>
<wicri:noRegion>China Medical University</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Xu, Hongtao" sort="Xu, Hongtao" uniqKey="Xu H" first="Hongtao" last="Xu">Hongtao Xu</name>
<affiliation wicri:level="1"><nlm:aff id="I3">College of Basic Medical Sciences, China Medical University, China</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea>College of Basic Medical Sciences, China Medical University</wicri:regionArea>
<wicri:noRegion>China Medical University</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Chen, Shimin" sort="Chen, Shimin" uniqKey="Chen S" first="Shimin" last="Chen">Shimin Chen</name>
<affiliation wicri:level="1"><nlm:aff id="I1">School of Public Health, China Medical University, China</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea>School of Public Health, China Medical University</wicri:regionArea>
<wicri:noRegion>China Medical University</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Zhao, Rui" sort="Zhao, Rui" uniqKey="Zhao R" first="Rui" last="Zhao">Rui Zhao</name>
<affiliation wicri:level="1"><nlm:aff id="I4">School of Forensic Medicine, China Medical University, China</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea>School of Forensic Medicine, China Medical University</wicri:regionArea>
<wicri:noRegion>China Medical University</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Pi, Jingbo" sort="Pi, Jingbo" uniqKey="Pi J" first="Jingbo" last="Pi">Jingbo Pi</name>
<affiliation wicri:level="1"><nlm:aff id="I1">School of Public Health, China Medical University, China</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea>School of Public Health, China Medical University</wicri:regionArea>
<wicri:noRegion>China Medical University</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Xu, Yuanyuan" sort="Xu, Yuanyuan" uniqKey="Xu Y" first="Yuanyuan" last="Xu">Yuanyuan Xu</name>
<affiliation wicri:level="1"><nlm:aff id="I1">School of Public Health, China Medical University, China</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea>School of Public Health, China Medical University</wicri:regionArea>
<wicri:noRegion>China Medical University</wicri:noRegion>
</affiliation>
</author>
</analytic>
<series><title level="j">Oxidative Medicine and Cellular Longevity</title>
<idno type="ISSN">1942-0900</idno>
<idno type="eISSN">1942-0994</idno>
<imprint><date when="2019">2019</date>
</imprint>
</series>
</biblStruct>
</sourceDesc>
</fileDesc>
<profileDesc><textClass></textClass>
</profileDesc>
</teiHeader>
<front><div type="abstract" xml:lang="en"><p>Chronic exposure to arsenic induces a variety of cancers, particularly in the skin. Autophagy is a highly conserved process which plays a dual role in tumorigenesis. In the present study, we found that chronic exposure to an environmentally relevant dose of arsenite induced malignant transformation of human keratinocytes (HaCaT) with dysregulated autophagy as indicated by an increased number of autophagosomes, activation of mTORC1 pathway, and elevated protein levels of p62 and LC3II. Meanwhile, arsenite-transformed cells showed lower intracellular levels of reactive oxygen species compared with control. Silencing <italic>p62</italic>
ameliorated elevation in mRNA levels of NRF2 downstream genes (<italic>AKR1C1</italic>
and <italic>NQO1</italic>
) and malignant phenotypes (acquired invasiveness and anchor-independent growth) induced by chronic arsenite exposure. On the other hand, silencing <italic>NRF2</italic>
abrogated the increase in mRNA and protein levels of p62 and malignant phenotypes induced by arsenite. In response to acute arsenite exposure, impaired autophagic flux with an increase in p62 protein level and interrupted autophagosome-lysosome fusion was observed. The increase in p62 protein levels in response to arsenite was not completely dependent on NRF2 activation and at least partially attributed to protein degradation. Our data indicate that accumulation of p62 by impaired autophagic flux is involved in the activation of NRF2 and contributes to skin tumorigenesis due to chronic arsenite exposure.</p>
</div>
</front>
<back><div1 type="bibliography"><listBibl><biblStruct></biblStruct>
<biblStruct><analytic><author><name sortKey="Platanias, L C" uniqKey="Platanias L">L. C. Platanias</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Alain, G" uniqKey="Alain G">G. Alain</name>
</author>
<author><name sortKey="Tousignant, J" uniqKey="Tousignant J">J. Tousignant</name>
</author>
<author><name sortKey="Rozenfarb, E" uniqKey="Rozenfarb E">E. Rozenfarb</name>
</author>
</analytic>
</biblStruct>
<biblStruct></biblStruct>
<biblStruct></biblStruct>
<biblStruct><analytic><author><name sortKey="Degenhardt, K" uniqKey="Degenhardt K">K. Degenhardt</name>
</author>
<author><name sortKey="Mathew, R" uniqKey="Mathew R">R. Mathew</name>
</author>
<author><name sortKey="Beaudoin, B" uniqKey="Beaudoin B">B. Beaudoin</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="White, E" uniqKey="White E">E. White</name>
</author>
<author><name sortKey="Karp, C" uniqKey="Karp C">C. Karp</name>
</author>
<author><name sortKey="Strohecker, A M" uniqKey="Strohecker A">A. M. Strohecker</name>
</author>
<author><name sortKey="Guo, Y" uniqKey="Guo Y">Y. Guo</name>
</author>
<author><name sortKey="Mathew, R" uniqKey="Mathew R">R. Mathew</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Rybstein, M D" uniqKey="Rybstein M">M. D. Rybstein</name>
</author>
<author><name sortKey="Bravo San Pedro, J M" uniqKey="Bravo San Pedro J">J. M. Bravo-San Pedro</name>
</author>
<author><name sortKey="Kroemer, G" uniqKey="Kroemer G">G. Kroemer</name>
</author>
<author><name sortKey="Galluzzi, L" uniqKey="Galluzzi L">L. Galluzzi</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Amaravadi, R K" uniqKey="Amaravadi R">R. K. Amaravadi</name>
</author>
<author><name sortKey="Thompson, C B" uniqKey="Thompson C">C. B. Thompson</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Rao, S" uniqKey="Rao S">S. Rao</name>
</author>
<author><name sortKey="Tortola, L" uniqKey="Tortola L">L. Tortola</name>
</author>
<author><name sortKey="Perlot, T" uniqKey="Perlot T">T. Perlot</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Rogov, V" uniqKey="Rogov V">V. Rogov</name>
</author>
<author><name sortKey="Dotsch, V" uniqKey="Dotsch V">V. Dotsch</name>
</author>
<author><name sortKey="Johansen, T" uniqKey="Johansen T">T. Johansen</name>
</author>
<author><name sortKey="Kirkin, V" uniqKey="Kirkin V">V. Kirkin</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Thompson, H G R" uniqKey="Thompson H">H. G. R. Thompson</name>
</author>
<author><name sortKey="Harris, J W" uniqKey="Harris J">J. W. Harris</name>
</author>
<author><name sortKey="Wold, B J" uniqKey="Wold B">B. J. Wold</name>
</author>
<author><name sortKey="Lin, F" uniqKey="Lin F">F. Lin</name>
</author>
<author><name sortKey="Brody, J P" uniqKey="Brody J">J. P. Brody</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Inoue, D" uniqKey="Inoue D">D. Inoue</name>
</author>
<author><name sortKey="Suzuki, T" uniqKey="Suzuki T">T. Suzuki</name>
</author>
<author><name sortKey="Mitsuishi, Y" uniqKey="Mitsuishi Y">Y. Mitsuishi</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Umemura, A" uniqKey="Umemura A">A. Umemura</name>
</author>
<author><name sortKey="He, F" uniqKey="He F">F. He</name>
</author>
<author><name sortKey="Taniguchi, K" uniqKey="Taniguchi K">K. Taniguchi</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Qiang, L" uniqKey="Qiang L">L. Qiang</name>
</author>
<author><name sortKey="Zhao, B" uniqKey="Zhao B">B. Zhao</name>
</author>
<author><name sortKey="Ming, M" uniqKey="Ming M">M. Ming</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Moscat, J" uniqKey="Moscat J">J. Moscat</name>
</author>
<author><name sortKey="Diaz Meco, M T" uniqKey="Diaz Meco M">M. T. Diaz-Meco</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Jain, A" uniqKey="Jain A">A. Jain</name>
</author>
<author><name sortKey="Lamark, T" uniqKey="Lamark T">T. Lamark</name>
</author>
<author><name sortKey="Sj Ttem, E" uniqKey="Sj Ttem E">E. Sjøttem</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Yamamoto, M" uniqKey="Yamamoto M">M. Yamamoto</name>
</author>
<author><name sortKey="Kensler, T W" uniqKey="Kensler T">T. W. Kensler</name>
</author>
<author><name sortKey="Motohashi, H" uniqKey="Motohashi H">H. Motohashi</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Komatsu, M" uniqKey="Komatsu M">M. Komatsu</name>
</author>
<author><name sortKey="Kurokawa, H" uniqKey="Kurokawa H">H. Kurokawa</name>
</author>
<author><name sortKey="Waguri, S" uniqKey="Waguri S">S. Waguri</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Lau, A" uniqKey="Lau A">A. Lau</name>
</author>
<author><name sortKey="Wang, X J" uniqKey="Wang X">X. J. Wang</name>
</author>
<author><name sortKey="Zhao, F" uniqKey="Zhao F">F. Zhao</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Pi, J" uniqKey="Pi J">J. Pi</name>
</author>
<author><name sortKey="Diwan, B A" uniqKey="Diwan B">B. A. Diwan</name>
</author>
<author><name sortKey="Sun, Y" uniqKey="Sun Y">Y. Sun</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Lau, A" uniqKey="Lau A">A. Lau</name>
</author>
<author><name sortKey="Zheng, Y" uniqKey="Zheng Y">Y. Zheng</name>
</author>
<author><name sortKey="Tao, S" uniqKey="Tao S">S. Tao</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Qi, Y" uniqKey="Qi Y">Y. Qi</name>
</author>
<author><name sortKey="Zhang, M" uniqKey="Zhang M">M. Zhang</name>
</author>
<author><name sortKey="Li, H" uniqKey="Li H">H. Li</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Liu, X" uniqKey="Liu X">X. Liu</name>
</author>
<author><name sortKey="Ling, M" uniqKey="Ling M">M. Ling</name>
</author>
<author><name sortKey="Chen, C" uniqKey="Chen C">C. Chen</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Shah, P" uniqKey="Shah P">P. Shah</name>
</author>
<author><name sortKey="Trinh, E" uniqKey="Trinh E">E. Trinh</name>
</author>
<author><name sortKey="Qiang, L" uniqKey="Qiang L">L. Qiang</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Dodson, M" uniqKey="Dodson M">M. Dodson</name>
</author>
<author><name sortKey="De La Vega, M R" uniqKey="De La Vega M">M. R. de la Vega</name>
</author>
<author><name sortKey="Harder, B" uniqKey="Harder B">B. Harder</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Leone, A" uniqKey="Leone A">A. Leone</name>
</author>
<author><name sortKey="Flatow, U" uniqKey="Flatow U">U. Flatow</name>
</author>
<author><name sortKey="King, C R" uniqKey="King C">C. R. King</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Valko, M" uniqKey="Valko M">M. Valko</name>
</author>
<author><name sortKey="Morris, H" uniqKey="Morris H">H. Morris</name>
</author>
<author><name sortKey="Cronin, M T" uniqKey="Cronin M">M. T. Cronin</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Bj Rk Y, G" uniqKey="Bj Rk Y G">G. Bjørkøy</name>
</author>
<author><name sortKey="Lamark, T" uniqKey="Lamark T">T. Lamark</name>
</author>
<author><name sortKey="Pankiv, S" uniqKey="Pankiv S">S. Pankiv</name>
</author>
<author><name sortKey=" Vervatn, A" uniqKey=" Vervatn A">A. Øvervatn</name>
</author>
<author><name sortKey="Brech, A" uniqKey="Brech A">A. Brech</name>
</author>
<author><name sortKey="Johansen, T" uniqKey="Johansen T">T. Johansen</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Mizushima, N" uniqKey="Mizushima N">N. Mizushima</name>
</author>
<author><name sortKey="Yoshimori, T" uniqKey="Yoshimori T">T. Yoshimori</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Shintani, T" uniqKey="Shintani T">T. Shintani</name>
</author>
<author><name sortKey="Klionsky, D J" uniqKey="Klionsky D">D. J. Klionsky</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Pajares, M" uniqKey="Pajares M">M. Pajares</name>
</author>
<author><name sortKey="Rojo, A I" uniqKey="Rojo A">A. I. Rojo</name>
</author>
<author><name sortKey="Arias, E" uniqKey="Arias E">E. Arias</name>
</author>
<author><name sortKey="Diaz Carretero, A" uniqKey="Diaz Carretero A">A. Diaz-Carretero</name>
</author>
<author><name sortKey="Cuervo, A M" uniqKey="Cuervo A">A. M. Cuervo</name>
</author>
<author><name sortKey="Cuadrado, A" uniqKey="Cuadrado A">A. Cuadrado</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Pi, J" uniqKey="Pi J">J. Pi</name>
</author>
<author><name sortKey="He, Y" uniqKey="He Y">Y. He</name>
</author>
<author><name sortKey="Bortner, C" uniqKey="Bortner C">C. Bortner</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Zhao, R" uniqKey="Zhao R">R. Zhao</name>
</author>
<author><name sortKey="Hou, Y" uniqKey="Hou Y">Y. Hou</name>
</author>
<author><name sortKey="Zhang, Q" uniqKey="Zhang Q">Q. Zhang</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Zhao, R" uniqKey="Zhao R">R. Zhao</name>
</author>
<author><name sortKey="Yang, B" uniqKey="Yang B">B. Yang</name>
</author>
<author><name sortKey="Wang, L" uniqKey="Wang L">L. Wang</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Ke, N" uniqKey="Ke N">N. Ke</name>
</author>
<author><name sortKey="Wang, X" uniqKey="Wang X">X. Wang</name>
</author>
<author><name sortKey="Xu, X" uniqKey="Xu X">X. Xu</name>
</author>
<author><name sortKey="Abassi, Y A" uniqKey="Abassi Y">Y. A. Abassi</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Xu, Y" uniqKey="Xu Y">Y. Xu</name>
</author>
<author><name sortKey="Tokar, E J" uniqKey="Tokar E">E. J. Tokar</name>
</author>
<author><name sortKey="Sun, Y" uniqKey="Sun Y">Y. Sun</name>
</author>
<author><name sortKey="Waalkes, M P" uniqKey="Waalkes M">M. P. Waalkes</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Ghosh, P" uniqKey="Ghosh P">P. Ghosh</name>
</author>
<author><name sortKey="Banerjee, M" uniqKey="Banerjee M">M. Banerjee</name>
</author>
<author><name sortKey="De Chaudhuri, S" uniqKey="De Chaudhuri S">S. de Chaudhuri</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Tseng, W P" uniqKey="Tseng W">W. P. Tseng</name>
</author>
<author><name sortKey="Chu, H M" uniqKey="Chu H">H. M. Chu</name>
</author>
<author><name sortKey="How, S W" uniqKey="How S">S. W. How</name>
</author>
<author><name sortKey="Fong, J M" uniqKey="Fong J">J. M. Fong</name>
</author>
<author><name sortKey="Lin, C S" uniqKey="Lin C">C. S. Lin</name>
</author>
<author><name sortKey="Yeh, S" uniqKey="Yeh S">S. Yeh</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Edwards, D H" uniqKey="Edwards D">D. H. Edwards</name>
</author>
<author><name sortKey="Li, Y" uniqKey="Li Y">Y. Li</name>
</author>
<author><name sortKey="Ellinsworth, D C" uniqKey="Ellinsworth D">D. C. Ellinsworth</name>
</author>
<author><name sortKey="Griffith, T M" uniqKey="Griffith T">T. M. Griffith</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Snow, E T" uniqKey="Snow E">E. T. Snow</name>
</author>
<author><name sortKey="Sykora, P" uniqKey="Sykora P">P. Sykora</name>
</author>
<author><name sortKey="Durham, T" uniqKey="Durham T">T. Durham</name>
</author>
<author><name sortKey="Klein, C" uniqKey="Klein C">C. Klein</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Zhang, D D" uniqKey="Zhang D">D. D. Zhang</name>
</author>
<author><name sortKey="Hannink, M" uniqKey="Hannink M">M. Hannink</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Son, Y O" uniqKey="Son Y">Y. O. Son</name>
</author>
<author><name sortKey="Pratheeshkumar, P" uniqKey="Pratheeshkumar P">P. Pratheeshkumar</name>
</author>
<author><name sortKey="Roy, R V" uniqKey="Roy R">R. V. Roy</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Ramos Gomez, M" uniqKey="Ramos Gomez M">M. Ramos-Gomez</name>
</author>
<author><name sortKey="Kwak, M K" uniqKey="Kwak M">M. K. Kwak</name>
</author>
<author><name sortKey="Dolan, P M" uniqKey="Dolan P">P. M. Dolan</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Ramos Gomez, M" uniqKey="Ramos Gomez M">M. Ramos-Gomez</name>
</author>
<author><name sortKey="Dolan, P M" uniqKey="Dolan P">P. M. Dolan</name>
</author>
<author><name sortKey="Itoh, K" uniqKey="Itoh K">K. Itoh</name>
</author>
<author><name sortKey="Yamamoto, M" uniqKey="Yamamoto M">M. Yamamoto</name>
</author>
<author><name sortKey="Kensler, T W" uniqKey="Kensler T">T. W. Kensler</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Rolfs, F" uniqKey="Rolfs F">F. Rolfs</name>
</author>
<author><name sortKey="Huber, M" uniqKey="Huber M">M. Huber</name>
</author>
<author><name sortKey="Kuehne, A" uniqKey="Kuehne A">A. Kuehne</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Ni, H M" uniqKey="Ni H">H. M. Ni</name>
</author>
<author><name sortKey="Woolbright, B L" uniqKey="Woolbright B">B. L. Woolbright</name>
</author>
<author><name sortKey="Williams, J" uniqKey="Williams J">J. Williams</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Ngo, H K C" uniqKey="Ngo H">H. K. C. Ngo</name>
</author>
<author><name sortKey="Kim, D H" uniqKey="Kim D">D. H. Kim</name>
</author>
<author><name sortKey="Cha, Y N" uniqKey="Cha Y">Y. N. Cha</name>
</author>
<author><name sortKey="Na, H K" uniqKey="Na H">H. K. Na</name>
</author>
<author><name sortKey="Surh, Y J" uniqKey="Surh Y">Y. J. Surh</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Fu, J" uniqKey="Fu J">J. Fu</name>
</author>
<author><name sortKey="Woods, C G" uniqKey="Woods C">C. G. Woods</name>
</author>
<author><name sortKey="Yehuda Shnaidman, E" uniqKey="Yehuda Shnaidman E">E. Yehuda-Shnaidman</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Copple, I M" uniqKey="Copple I">I. M. Copple</name>
</author>
<author><name sortKey="Lister, A" uniqKey="Lister A">A. Lister</name>
</author>
<author><name sortKey="Obeng, A D" uniqKey="Obeng A">A. D. Obeng</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Taguchi, K" uniqKey="Taguchi K">K. Taguchi</name>
</author>
<author><name sortKey="Fujikawa, N" uniqKey="Fujikawa N">N. Fujikawa</name>
</author>
<author><name sortKey="Komatsu, M" uniqKey="Komatsu M">M. Komatsu</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Komatsu, M" uniqKey="Komatsu M">M. Komatsu</name>
</author>
<author><name sortKey="Kageyama, S" uniqKey="Kageyama S">S. Kageyama</name>
</author>
<author><name sortKey="Ichimura, Y" uniqKey="Ichimura Y">Y. Ichimura</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Linares, J F" uniqKey="Linares J">J. F. Linares</name>
</author>
<author><name sortKey="Duran, A" uniqKey="Duran A">A. Duran</name>
</author>
<author><name sortKey="Reina Campos, M" uniqKey="Reina Campos M">M. Reina-Campos</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Eskelinen, E L" uniqKey="Eskelinen E">E. L. Eskelinen</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Issa, A R" uniqKey="Issa A">A. R. Issa</name>
</author>
<author><name sortKey="Sun, J" uniqKey="Sun J">J. Sun</name>
</author>
<author><name sortKey="Petitgas, C" uniqKey="Petitgas C">C. Petitgas</name>
</author>
</analytic>
</biblStruct>
</listBibl>
</div1>
</back>
</TEI>
<pmc article-type="research-article"><pmc-dir>properties open_access</pmc-dir>
<front><journal-meta><journal-id journal-id-type="nlm-ta">Oxid Med Cell Longev</journal-id>
<journal-id journal-id-type="iso-abbrev">Oxid Med Cell Longev</journal-id>
<journal-id journal-id-type="publisher-id">OMCL</journal-id>
<journal-title-group><journal-title>Oxidative Medicine and Cellular Longevity</journal-title>
</journal-title-group>
<issn pub-type="ppub">1942-0900</issn>
<issn pub-type="epub">1942-0994</issn>
<publisher><publisher-name>Hindawi</publisher-name>
</publisher>
</journal-meta>
<article-meta><article-id pub-id-type="pmid">31781319</article-id>
<article-id pub-id-type="pmc">6875345</article-id>
<article-id pub-id-type="doi">10.1155/2019/1038932</article-id>
<article-categories><subj-group subj-group-type="heading"><subject>Research Article</subject>
</subj-group>
</article-categories>
<title-group><article-title>Enhanced p62-NRF2 Feedback Loop due to Impaired Autophagic Flux Contributes to Arsenic-Induced Malignant Transformation of Human Keratinocytes</article-title>
</title-group>
<contrib-group><contrib contrib-type="author"><name><surname>Wu</surname>
<given-names>Xiafang</given-names>
</name>
<xref ref-type="aff" rid="I1"><sup>1</sup>
</xref>
<xref ref-type="aff" rid="I2"><sup>2</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Sun</surname>
<given-names>Ru</given-names>
</name>
<xref ref-type="aff" rid="I1"><sup>1</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Wang</surname>
<given-names>Huihui</given-names>
</name>
<xref ref-type="aff" rid="I1"><sup>1</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Yang</surname>
<given-names>Bei</given-names>
</name>
<xref ref-type="aff" rid="I3"><sup>3</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Wang</surname>
<given-names>Fang</given-names>
</name>
<xref ref-type="aff" rid="I1"><sup>1</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Xu</surname>
<given-names>Hongtao</given-names>
</name>
<xref ref-type="aff" rid="I3"><sup>3</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Chen</surname>
<given-names>Shimin</given-names>
</name>
<xref ref-type="aff" rid="I1"><sup>1</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Zhao</surname>
<given-names>Rui</given-names>
</name>
<xref ref-type="aff" rid="I4"><sup>4</sup>
</xref>
</contrib>
<contrib contrib-type="author"><contrib-id contrib-id-type="orcid" authenticated="false">https://orcid.org/0000-0003-0227-8041</contrib-id>
<name><surname>Pi</surname>
<given-names>Jingbo</given-names>
</name>
<xref ref-type="aff" rid="I1"><sup>1</sup>
</xref>
</contrib>
<contrib contrib-type="author" corresp="yes"><contrib-id contrib-id-type="orcid" authenticated="false">https://orcid.org/0000-0003-2354-9453</contrib-id>
<name><surname>Xu</surname>
<given-names>Yuanyuan</given-names>
</name>
<email>yyxu@cmu.edu.cn</email>
<xref ref-type="aff" rid="I1"><sup>1</sup>
</xref>
</contrib>
</contrib-group>
<aff id="I1"><sup>1</sup>
School of Public Health, China Medical University, China</aff>
<aff id="I2"><sup>2</sup>
The First Hospital of China Medical University, China</aff>
<aff id="I3"><sup>3</sup>
College of Basic Medical Sciences, China Medical University, China</aff>
<aff id="I4"><sup>4</sup>
School of Forensic Medicine, China Medical University, China</aff>
<author-notes><fn fn-type="other"><p>Academic Editor: Aldrin V. Gomes</p>
</fn>
</author-notes>
<pub-date pub-type="collection"><year>2019</year>
</pub-date>
<pub-date pub-type="epub"><day>30</day>
<month>10</month>
<year>2019</year>
</pub-date>
<volume>2019</volume>
<elocation-id>1038932</elocation-id>
<history><date date-type="received"><day>13</day>
<month>3</month>
<year>2019</year>
</date>
<date date-type="rev-recd"><day>16</day>
<month>8</month>
<year>2019</year>
</date>
<date date-type="accepted"><day>29</day>
<month>8</month>
<year>2019</year>
</date>
</history>
<permissions><copyright-statement>Copyright © 2019 Xiafang Wu et al.</copyright-statement>
<copyright-year>2019</copyright-year>
<license xlink:href="http://creativecommons.org/licenses/by/4.0/"><license-p>This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.</license-p>
</license>
</permissions>
<abstract><p>Chronic exposure to arsenic induces a variety of cancers, particularly in the skin. Autophagy is a highly conserved process which plays a dual role in tumorigenesis. In the present study, we found that chronic exposure to an environmentally relevant dose of arsenite induced malignant transformation of human keratinocytes (HaCaT) with dysregulated autophagy as indicated by an increased number of autophagosomes, activation of mTORC1 pathway, and elevated protein levels of p62 and LC3II. Meanwhile, arsenite-transformed cells showed lower intracellular levels of reactive oxygen species compared with control. Silencing <italic>p62</italic>
ameliorated elevation in mRNA levels of NRF2 downstream genes (<italic>AKR1C1</italic>
and <italic>NQO1</italic>
) and malignant phenotypes (acquired invasiveness and anchor-independent growth) induced by chronic arsenite exposure. On the other hand, silencing <italic>NRF2</italic>
abrogated the increase in mRNA and protein levels of p62 and malignant phenotypes induced by arsenite. In response to acute arsenite exposure, impaired autophagic flux with an increase in p62 protein level and interrupted autophagosome-lysosome fusion was observed. The increase in p62 protein levels in response to arsenite was not completely dependent on NRF2 activation and at least partially attributed to protein degradation. Our data indicate that accumulation of p62 by impaired autophagic flux is involved in the activation of NRF2 and contributes to skin tumorigenesis due to chronic arsenite exposure.</p>
</abstract>
<funding-group><award-group><funding-source>Department of Science and Technology of Liaoning Province</funding-source>
</award-group>
<award-group><funding-source>Liaoning Revitalization Talents Program</funding-source>
<award-id>XLYC1807225</award-id>
</award-group>
<award-group><funding-source>National Natural Science Foundation of China</funding-source>
<award-id>81302391</award-id>
<award-id>81573187</award-id>
</award-group>
</funding-group>
</article-meta>
</front>
<floats-group><fig id="fig1" orientation="portrait" position="float"><label>Figure 1</label>
<caption><p>Alterations of autophagy markers and upstream signaling pathways in arsenite-transformed (As-TM) cells. HaCaT cells were treated with 100 nM of sodium arsenite (As) for 30 weeks. Passage-matched nontreated cells were used as the control (Con). (a) Cell invasion analyzed with real-time cell analysis (RTCA) xCELLigence system. The left chart shows the kinetic analysis of cell invasion. The right panel shows the average index reflecting cell invasion capacity at 45 h. (b) Colony formation in soft agar. Representative images of the control (upper left panel), As-TM cells (upper right panel), and quantification of colonies (lower panel) are shown. Scale bar is 100 <italic>μ</italic>
m. For quantification, three fields of view were randomly selected from each 35 mm culture dish. The number of clones with a diameter greater than 100 <italic>μ</italic>
m was counted. (c) Increase in number of autophagosomes in As-TM cells. Autophagosomes were observed with transmission electron microscopy (TEM). Scale bar is 2 <italic>μ</italic>
m (up) and 0.5 <italic>μ</italic>
m (down). Arrows indicate autophagosomes. (d) Number of autophagosomes per cell according to TEM. (e) Western blot for autophagy markers, LC3 and p62. Upper: representative image; lower: quantification of protein levels of LC3II/I and p62 determined with Western blot. (f) Western blot for BECN1 and proteins in the mTORC1 pathway, including p-mTOR, mTOR, RAPTOR, p-P70S6K, and P70S6K. Upper: representative image; lower: quantification of p-mTOR/mTOR and p-P70S6K/P70S6K determined with Western blot. <italic>n</italic>
= 3 except for colony formation, in which <italic>n</italic>
= 6. <sup>∗</sup>
<italic>p</italic>
< 0.05, compared with Con.</p>
</caption>
<graphic xlink:href="OMCL2019-1038932.001"></graphic>
</fig>
<fig id="fig2" orientation="portrait" position="float"><label>Figure 2</label>
<caption><p>Long-term exposure to low-level arsenite induced adaptive antioxidative response in HaCaT cells. (a) Representative histogram for intracellular ROS levels detected by flow cytometer. As-TM cells or passage-matched nontreated control (Con) was challenged with 10 <italic>μ</italic>
M of sodium arsenite (As) or equal volume of PBS (Veh) for 24 h. (b) Quantification of intracellular ROS levels determined by flow cytometer. (c) mRNA levels of NRF2 and p62 in As-TM and control cells. (d) Western blot of NRF2 in As-TM and control cells. Upper: representative image; lower: quantification of NRF2 protein levels determined with Western blot. <italic>n</italic>
= 3. <sup>∗</sup>
<italic>p</italic>
< 0.05, compared with Con compartment. <sup>#</sup>
<italic>p</italic>
< 0.05, compared with Veh compartment.</p>
</caption>
<graphic xlink:href="OMCL2019-1038932.002"></graphic>
</fig>
<fig id="fig3" orientation="portrait" position="float"><label>Figure 3</label>
<caption><p>Amplification of p62-NRF2 feedback loop is required for the acquisition of arsenite-induced malignant phenotypes. (a) mRNA levels of <italic>p62</italic>
in HaCaT cells infected with lentiviral vector expressing shRNA targeting <italic>p62</italic>
(<italic>p62</italic>
-KD) or scrambled nontarget negative control (SCR). (b) Protein levels of p62 and NRF2 in <italic>p62</italic>
-KD and SCR cells. Left: representative image; right: quantification of protein levels of p62 and NRF2 determined with Western blot. (c) mRNA levels of <italic>NRF2</italic>
downstream genes, <italic>AKR1C1</italic>
, <italic>GCLC</italic>
, and <italic>NQO1</italic>
, in <italic>p62</italic>
-KD and SCR cells. (d) mRNA levels of <italic>NRF2</italic>
and its downstream genes in chronic arsenite-exposed cells with <italic>NRF2</italic>
knockdown (<italic>NRF2</italic>
-KD). (e) mRNA and protein levels of p62 in <italic>NRF2</italic>
-KD cells analyzed with RT-PCR (left) and Western blot (right), respectively. Upper right: representative image for Western blot; lower right: quantification of p62 protein levels determined with Western blot. (f) Invasion capacity determined by xCELLigence RTCA. Cell index at 45 h after seeding was used to assess invasion capacity. (g) Colony formation in soft agar. Representative image (upper) and quantification of the colonies (lower). Scale bar is 100 <italic>μ</italic>
m. As (As+): cells were chronically exposed to 100 nM of sodium arsenite for 30 weeks. Con: passage-matched nontreated cells. <italic>n</italic>
= 3 except for colony formation assay, in which <italic>n</italic>
= 6. <sup>∗</sup>
<italic>p</italic>
< 0.05, compared with control (As-) compartment. <sup>#</sup>
<italic>p</italic>
< 0.05, compared with SCR (<italic>NRF2</italic>
-KD- or <italic>p62</italic>
-KD-) compartment.</p>
</caption>
<graphic xlink:href="OMCL2019-1038932.003"></graphic>
</fig>
<fig id="fig4" orientation="portrait" position="float"><label>Figure 4</label>
<caption><p>Arsenite inhibits autophagosome-lysosome fusion. (a) Autophagosomes observed with TEM in cells nontreated (Con) or treated with 100 nM or 200 nM sodium arsenite for 4 h. Arrows indicate autophagosomes. Scale bar is 2 <italic>μ</italic>
m (up) and 0.5 <italic>μ</italic>
m (down). (b) Number of autophagosomes per cell according to TEM. (c) Western blot for LC3 and p62 in HaCaT cells treated with 100 nM, 200 nM, or 500 nM arsenite for 6 h. (d) Western blot for LC3 and p62 in HaCaT cells treated with 100 nM arsenite at different time points. (e) Protein levels of p62 detected with Western blot. Arsenite-induced inhibition of autophagic flux was tested with chloroquine (CQ, 30 <italic>μ</italic>
M) pretreatment and in the absence (-) or presence (+) of 100 nM arsenite for 6 h. (f) Quantification of orange/yellow LC3 puncta in the cell. HaCaT cells were transfected with a tandem mRFP-GFP-LC3 and then treated with 100 nM arsenite for 4 h or 30 <italic>μ</italic>
M CQ for 6 h. The number of puncta in cells was counted using ImageJ software. Average number of orange/yellow puncta per cell from 16 randomly selected cells in each group was shown. (g) Representative image of LC3 fluorescence observed by a confocal microscope. Scale bar is 50 <italic>μ</italic>
m. (h) Western blot for LAMP1 and LAMP2 in HaCaT cells treated with 100 nM, 200 nM, or 500 nM arsenite for 6 h. (i) Western blot for LAMP1 and LAMP2 in HaCaT cells exposed to 100 nM arsenite at different time points. For Western blot, upper: representative image; lower: quantification of protein levels determined with Western blot. <italic>n</italic>
= 3. <sup>∗</sup>
<italic>p</italic>
< 0.05, compared with Con (or As- and CQ-).</p>
</caption>
<graphic xlink:href="OMCL2019-1038932.004"></graphic>
</fig>
<fig id="fig5" orientation="portrait" position="float"><label>Figure 5</label>
<caption><p>Enhanced transcription and decreased protein turnover contribute to accumulation of p62 protein in response to arsenite exposure. (a) Western blot of NRF2 and p62 under basal and arsenite-treated conditions in SCR and <italic>NRF2</italic>
-KD cells. As: cells were treated with 100 nM sodium arsenite for 6 h. PC: positive control, cells were treated with 20 <italic>μ</italic>
M sodium arsenite for 6 h. (b) mRNA levels of <italic>p62</italic>
, <italic>NQO1</italic>
, and <italic>GCLC</italic>
in HaCaT cells treated with 100 nM sodium arsenite at different time points. (c) Exposure to arsenite inhibited p62 degradation in HaCaT cells. Cells were treated with CHX (10 <italic>μ</italic>
g/mL) or CHX+ As (100 nM) at different time points, followed by Western blot analysis. For Western blot, left: representative image; right: quantification of p62 protein levels. <italic>n</italic>
= 3. <sup>∗</sup>
<italic>p</italic>
< 0.05, compared with Con compartment. <sup>#</sup>
<italic>p</italic>
< 0.05, compared with SCR compartment or CHX-treated compartment.</p>
</caption>
<graphic xlink:href="OMCL2019-1038932.005"></graphic>
</fig>
</floats-group>
</pmc>
<affiliations><list><country><li>République populaire de Chine</li>
</country>
</list>
<tree><country name="République populaire de Chine"><noRegion><name sortKey="Wu, Xiafang" sort="Wu, Xiafang" uniqKey="Wu X" first="Xiafang" last="Wu">Xiafang Wu</name>
</noRegion>
<name sortKey="Chen, Shimin" sort="Chen, Shimin" uniqKey="Chen S" first="Shimin" last="Chen">Shimin Chen</name>
<name sortKey="Pi, Jingbo" sort="Pi, Jingbo" uniqKey="Pi J" first="Jingbo" last="Pi">Jingbo Pi</name>
<name sortKey="Sun, Ru" sort="Sun, Ru" uniqKey="Sun R" first="Ru" last="Sun">Ru Sun</name>
<name sortKey="Wang, Fang" sort="Wang, Fang" uniqKey="Wang F" first="Fang" last="Wang">Fang Wang</name>
<name sortKey="Wang, Huihui" sort="Wang, Huihui" uniqKey="Wang H" first="Huihui" last="Wang">Huihui Wang</name>
<name sortKey="Wu, Xiafang" sort="Wu, Xiafang" uniqKey="Wu X" first="Xiafang" last="Wu">Xiafang Wu</name>
<name sortKey="Xu, Hongtao" sort="Xu, Hongtao" uniqKey="Xu H" first="Hongtao" last="Xu">Hongtao Xu</name>
<name sortKey="Xu, Yuanyuan" sort="Xu, Yuanyuan" uniqKey="Xu Y" first="Yuanyuan" last="Xu">Yuanyuan Xu</name>
<name sortKey="Yang, Bei" sort="Yang, Bei" uniqKey="Yang B" first="Bei" last="Yang">Bei Yang</name>
<name sortKey="Zhao, Rui" sort="Zhao, Rui" uniqKey="Zhao R" first="Rui" last="Zhao">Rui Zhao</name>
</country>
</tree>
</affiliations>
</record>
Pour manipuler ce document sous Unix (Dilib)
EXPLOR_STEP=$WICRI_ROOT/Sante/explor/ChloroquineV1/Data/Pmc/Checkpoint
HfdSelect -h $EXPLOR_STEP/biblio.hfd -nk 000778 | SxmlIndent | more
Ou
HfdSelect -h $EXPLOR_AREA/Data/Pmc/Checkpoint/biblio.hfd -nk 000778 | SxmlIndent | more
Pour mettre un lien sur cette page dans le réseau Wicri
{{Explor lien |wiki= Sante |area= ChloroquineV1 |flux= Pmc |étape= Checkpoint |type= RBID |clé= PMC:6875345 |texte= Enhanced p62-NRF2 Feedback Loop due to Impaired Autophagic Flux Contributes to Arsenic-Induced Malignant Transformation of Human Keratinocytes }}
Pour générer des pages wiki
HfdIndexSelect -h $EXPLOR_AREA/Data/Pmc/Checkpoint/RBID.i -Sk "pubmed:31781319" \ | HfdSelect -Kh $EXPLOR_AREA/Data/Pmc/Checkpoint/biblio.hfd \ | NlmPubMed2Wicri -a ChloroquineV1
![]() | This area was generated with Dilib version V0.6.33. | ![]() |