TY - JOUR
T1 - Depletion of keratin 8/18 modulates oncogenic potential by governing multiple signaling pathways
AU - Tiwari, Richa
AU - Sahu, Indrajit
AU - Soni, Bihari Lal
AU - Sathe, Gajanan J.
AU - Thapa, Pankaj
AU - Patel, Pavan
AU - Sinha, Shruti
AU - Vadivel, Chella Krishna
AU - Patel, Shweta
AU - Jamghare, Sayli Nitin
AU - Oak, Swapnil
AU - Thorat, Rahul
AU - Gowda, Harsha
AU - Vaidya, Milind M.
N1 - Funding Information:
We thank Dr Sorab N Dalal (ACTREC, TMC, India) for his generous gift HEK293-FT and pTU6 PURO vector. We also thank Dr Rita Mulherkar (ACTREC, TMC, India) for providing us A431 cell line as a kind gift. We thank Pierre A. Coulombe (Professor at Johns Hopkins, Baltimore, Maryland, USA) for providing us the K17 antibody as a kind gift. We thank Dr Neil D. Perkins for providing 3x?B ConA Luc vector and ConA Luc control vector. We also thank Ankit Jain and Kiran Kumar (Institute of Bioinformatics, Bangalore) for their generous help during data analysis. We thank Dr Tamar Ziv (The Smoler Proteomics Center, Department of Biology, Technion, Israel) for her contribution in quantitative proteomics data analysis. We thank Dr. Hunain Alam for providing pTU6 Puro ShRNA K8.2 construct and his guidance during the work. This work was supported by grant from Department of Biotechnology (Grant no. BT/PR7860/BRB/10/1222/2013). RT was supported by fellowship from ACTREC, TMC.
Funding Information:
We thank Dr Sorab N Dalal (ACTREC, TMC, India) for his generous gift HEK293-FT and pTU6 PURO vector. We also thank Dr Rita Mulherkar (ACTREC, TMC, India) for providing us A431 cell line as a kind gift. We thank Pierre A. Coulombe (Professor at Johns Hopkins, Baltimore, Maryland, USA) for providing us the K17 antibody as a kind gift. We thank Dr Neil D. Perkins for providing 3xκB ConA Luc vector and ConA Luc control vector. We also thank Ankit Jain and Kiran Kumar (Institute of Bioinformatics, Bangalore) for their generous help during data analysis. We thank Dr Tamar Ziv (The Smoler Proteomics Center, Department of Biology, Technion, Israel) for her contribution in quantitative proteomics data analysis. We thank Dr. Hunain Alam for providing pTU6 Puro ShRNA K8.2 construct and his guidance during the work. This work was supported by grant from Department of Biotechnology (Grant no. BT/PR7860/BRB/10/1222/2013). RT was supported by fellowship from ACTREC, TMC.
Publisher Copyright:
© 2018 Federation of European Biochemical Societies
Copyright:
Copyright 2019 Elsevier B.V., All rights reserved.
PY - 2018/4/12
Y1 - 2018/4/12
N2 - Keratin 8/18, the predominant keratin pair of simple epithelia, is often aberrantly expressed in various squamous cell carcinomas (SCCs) including skin SCC. Its aberrant expression is correlated with increased invasiveness and poor prognosis of the same, although the underlying mechanism is still unclear. A previous report from our laboratory has shown K8-mediated regulation of α6β4 integrin signaling and thereby tumorigenic potential of oral SCC-derived cells. Another study on transgenic mouse model has shown that during skin carcinogenesis, K8 favors conversion of papillomas toward malignancy. In order to understand the role of K8 and allied mechanism in skin SCC, K8 was stably knocked down in a skin epidermoid carcinoma-derived A431 cells. K8 downregulation significantly reduced the tumorigenic potential of these cells. In agreement with our phenotypic data, differential quantitative proteomics followed by IPA analysis showed altered expression of many proteins associated with biological functions including ‘Cancer’, ‘Cellular movement’, ‘Cell death and survival’, and ‘Cellular morphology’. Some of these proteins were TMS1, MARCKSL1, RanBP1, 14-3-3γ, Rho-GDI2, etc. Furthermore, to our surprise, there was a significant reduction in K17 protein stability upon loss of K8, probably due to its caspase-mediated degradation. This was supported by altered TMS1-NF-κB signaling, leading to increased apoptotic sensitivity of A431 cells which in turn affected ‘Cell death and survival’. Moreover, MARCKSL1-Paxillin1-Rac axis was found to be deregulated bestowing a possible mechanism behind altered ‘Cellular movement’ pathway. Altogether our study unravels a much broader regulatory role of K8, governing multiple signaling pathways and consequently regulating oncogenic potential of skin SCC-derived cells. Database: Proteome Xchange Consortium via PRIDE database (dataset identifier PXD007206).
AB - Keratin 8/18, the predominant keratin pair of simple epithelia, is often aberrantly expressed in various squamous cell carcinomas (SCCs) including skin SCC. Its aberrant expression is correlated with increased invasiveness and poor prognosis of the same, although the underlying mechanism is still unclear. A previous report from our laboratory has shown K8-mediated regulation of α6β4 integrin signaling and thereby tumorigenic potential of oral SCC-derived cells. Another study on transgenic mouse model has shown that during skin carcinogenesis, K8 favors conversion of papillomas toward malignancy. In order to understand the role of K8 and allied mechanism in skin SCC, K8 was stably knocked down in a skin epidermoid carcinoma-derived A431 cells. K8 downregulation significantly reduced the tumorigenic potential of these cells. In agreement with our phenotypic data, differential quantitative proteomics followed by IPA analysis showed altered expression of many proteins associated with biological functions including ‘Cancer’, ‘Cellular movement’, ‘Cell death and survival’, and ‘Cellular morphology’. Some of these proteins were TMS1, MARCKSL1, RanBP1, 14-3-3γ, Rho-GDI2, etc. Furthermore, to our surprise, there was a significant reduction in K17 protein stability upon loss of K8, probably due to its caspase-mediated degradation. This was supported by altered TMS1-NF-κB signaling, leading to increased apoptotic sensitivity of A431 cells which in turn affected ‘Cell death and survival’. Moreover, MARCKSL1-Paxillin1-Rac axis was found to be deregulated bestowing a possible mechanism behind altered ‘Cellular movement’ pathway. Altogether our study unravels a much broader regulatory role of K8, governing multiple signaling pathways and consequently regulating oncogenic potential of skin SCC-derived cells. Database: Proteome Xchange Consortium via PRIDE database (dataset identifier PXD007206).
KW - cell death and survival
KW - cellular movement
KW - MARCKSL1
KW - quantitative proteomics
KW - TMS1
UR - http://www.scopus.com/inward/record.url?scp=85042539317&partnerID=8YFLogxK
U2 - 10.1111/febs.14401
DO - 10.1111/febs.14401
M3 - Article
C2 - 29427328
AN - SCOPUS:85042539317
SN - 1742-464X
VL - 285
SP - 1251
EP - 1276
JO - FEBS Journal
JF - FEBS Journal
IS - 7
ER -
