Rab8 GTPase regulates Klotho-mediated inhibition of cell growth and progression by directly modulating its surface expression in human non-small cell lung cancer.
Animals
Biomarkers, Tumor
/ metabolism
Carcinogenesis
/ genetics
Carcinoma, Non-Small-Cell Lung
/ genetics
Cell Line, Tumor
Cell Membrane
/ metabolism
Cell Proliferation
Disease Progression
Down-Regulation
/ genetics
Endocytosis
Epithelial-Mesenchymal Transition
/ genetics
Gene Expression Regulation, Neoplastic
Glucuronidase
/ genetics
Humans
Klotho Proteins
Lung Neoplasms
/ genetics
Male
Mice, Inbred BALB C
Mice, Nude
Protein Binding
Survival Analysis
Wnt Signaling Pathway
/ genetics
rab GTP-Binding Proteins
/ genetics
Cell phenotype
Klotho
Non-small cell lung cancer
Rab8
Surface expression
Wnt signaling
Journal
EBioMedicine
ISSN: 2352-3964
Titre abrégé: EBioMedicine
Pays: Netherlands
ID NLM: 101647039
Informations de publication
Date de publication:
Nov 2019
Nov 2019
Historique:
received:
21
06
2019
revised:
18
10
2019
accepted:
22
10
2019
pubmed:
11
11
2019
medline:
1
4
2020
entrez:
11
11
2019
Statut:
ppublish
Résumé
The klotho (KL) gene is an anti-aging gene that has recently been shown to also function as a general tumor suppressor. However, there is currently only limited information regarding the potential molecular signals for regulation of Klotho without identifying precise molecular mechanisms or interactions. We performed a mass spectrometry (MS) assay to screen candidate proteins complexed with Klotho derived from immunoprecipitation in human non-small cell lung cancer (NSCLC) cells, and identified Rab8 to be the protein that most prominently interacts with Klotho. We further investigated whether Rab8 can regulate trafficking of Klotho and which process it would modulate using surface biotinylation assay, immunofluorescence and fluorescence ratio microscopy. Furthermore, we explored whether Rab8 is involved in Klotho-mediated function in NSCLC, and verified the results which we found in vivo using xenograft mouse model. We report discovery of Rab8 as a Klotho-interacting protein that acts as a critical modulator of Klotho surface expression in human NSCLC. In particular, we report that Rab8 is co-localized and associated with Klotho, and Klotho trafficking is regulated by Rab8. Moreover, we found that Rab8 modulates surface levels of Klotho via a post-biosynthetic pathway, as opposed to an endocytic pathway. Furthermore, we demonstrate that Rab8 is involved in Klotho-mediated regulation of cell proliferation, migration, invasiveness, epithelial-mesenchymal transition (EMT), and Wnt-β-catenin signaling in NSCLC. Additionally, Rab8 overexpression was also found to increase Klotho-mediated inhibition of NSCLC tumorigenesis in vivo. Overall, our findings suggest that Rab8 GTPase can regulate Klotho-mediated inhibition of Wnt signaling activity by modulating translocation of Klotho onto the cell surface, which in turn affects Klotho-mediated inhibition of cell proliferation, migration and invasiveness in NSCLC. These results have important implications for the development of new therapeutic targets, Klotho-related research in the context of NSCLC as well as other areas, and provide a working model for Rab8 function in the context of cancer and cancer biology.
Sections du résumé
BACKGROUND
BACKGROUND
The klotho (KL) gene is an anti-aging gene that has recently been shown to also function as a general tumor suppressor. However, there is currently only limited information regarding the potential molecular signals for regulation of Klotho without identifying precise molecular mechanisms or interactions.
METHODS
METHODS
We performed a mass spectrometry (MS) assay to screen candidate proteins complexed with Klotho derived from immunoprecipitation in human non-small cell lung cancer (NSCLC) cells, and identified Rab8 to be the protein that most prominently interacts with Klotho. We further investigated whether Rab8 can regulate trafficking of Klotho and which process it would modulate using surface biotinylation assay, immunofluorescence and fluorescence ratio microscopy. Furthermore, we explored whether Rab8 is involved in Klotho-mediated function in NSCLC, and verified the results which we found in vivo using xenograft mouse model.
FINDINGS
RESULTS
We report discovery of Rab8 as a Klotho-interacting protein that acts as a critical modulator of Klotho surface expression in human NSCLC. In particular, we report that Rab8 is co-localized and associated with Klotho, and Klotho trafficking is regulated by Rab8. Moreover, we found that Rab8 modulates surface levels of Klotho via a post-biosynthetic pathway, as opposed to an endocytic pathway. Furthermore, we demonstrate that Rab8 is involved in Klotho-mediated regulation of cell proliferation, migration, invasiveness, epithelial-mesenchymal transition (EMT), and Wnt-β-catenin signaling in NSCLC. Additionally, Rab8 overexpression was also found to increase Klotho-mediated inhibition of NSCLC tumorigenesis in vivo.
INTERPRETATION
CONCLUSIONS
Overall, our findings suggest that Rab8 GTPase can regulate Klotho-mediated inhibition of Wnt signaling activity by modulating translocation of Klotho onto the cell surface, which in turn affects Klotho-mediated inhibition of cell proliferation, migration and invasiveness in NSCLC. These results have important implications for the development of new therapeutic targets, Klotho-related research in the context of NSCLC as well as other areas, and provide a working model for Rab8 function in the context of cancer and cancer biology.
Identifiants
pubmed: 31707148
pii: S2352-3964(19)30710-8
doi: 10.1016/j.ebiom.2019.10.040
pmc: PMC6945242
pii:
doi:
Substances chimiques
Biomarkers, Tumor
0
Glucuronidase
EC 3.2.1.31
Klotho Proteins
EC 3.2.1.31
RAB8A protein, human
EC 3.6.1.-.
rab GTP-Binding Proteins
EC 3.6.5.2
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
118-132Informations de copyright
Copyright © 2019 The Authors. Published by Elsevier B.V. All rights reserved.
Références
Oncol Lett. 2017 Nov;14(5):5526-5532
pubmed: 29142604
EMBO J. 2007 Mar 21;26(6):1499-510
pubmed: 17332756
J Exp Clin Cancer Res. 2010 Jul 19;29:99
pubmed: 20642846
Lung Cancer. 2011 Jun;72(3):355-9
pubmed: 21075474
Neuron. 2003 Jul 3;39(1):69-84
pubmed: 12848933
J Cell Sci. 2006 Dec 1;119(Pt 23):4866-77
pubmed: 17105768
Mol Cancer. 2010 May 18;9:109
pubmed: 20482749
Proc Natl Acad Sci U S A. 2007 Dec 11;104(50):19796-801
pubmed: 18056631
Oncol Rep. 2012 Sep;28(3):1022-8
pubmed: 22710352
Nature. 2006 Dec 7;444(7120):770-4
pubmed: 17086194
Biochem J. 2009 Feb 15;418(1):163-72
pubmed: 18983266
Small GTPases. 2018 Mar 4;9(1-2):158-181
pubmed: 29239692
Cancer Lett. 2015 Jul 1;362(2):149-57
pubmed: 25827069
Mol Cancer Res. 2015 Oct;13(10):1398-407
pubmed: 26113466
Cytoskeleton (Hoboken). 2011 Oct;68(10):527-39
pubmed: 21850707
J Neurosci. 2013 May 22;33(21):9214-30
pubmed: 23699532
Vitam Horm. 2016;101:197-214
pubmed: 27125743
J Biol Chem. 2009 May 29;284(22):15126-36
pubmed: 19351881
Oncogene. 2008 Nov 27;27(56):7094-105
pubmed: 18762812
Endocr Rev. 2015 Apr;36(2):174-93
pubmed: 25695404
CA Cancer J Clin. 2018 Jan;68(1):7-30
pubmed: 29313949
Nat Rev Mol Cell Biol. 2002 Aug;3(8):600-14
pubmed: 12154371
Nat Commun. 2015 Apr 29;6:7035
pubmed: 25923845
J Biol Chem. 2001 Apr 20;276(16):13209-16
pubmed: 11278749
J Biomed Sci. 2016 Oct 6;23(1):70
pubmed: 27716280
Clin Cancer Res. 2011 Jul 1;17(13):4254-66
pubmed: 21571866
Int J Oncol. 2014 Aug;45(2):611-8
pubmed: 24818842
Cells. 2019 Aug 16;8(8):
pubmed: 31426400
Cancer Res. 2005 Apr 1;65(7):2516-9
pubmed: 15805241
PLoS One. 2013;8(2):e57391
pubmed: 23437382
FEBS Lett. 2009 Oct 6;583(19):3221-4
pubmed: 19737556
Cancer Treat Rev. 2018 Jan;62:50-60
pubmed: 29169144
J Cell Biol. 2001 Apr 30;153(3):555-68
pubmed: 11331306
Lancet. 2011 Nov 12;378(9804):1727-40
pubmed: 21565398
Curr Protein Pept Sci. 2014;15(8):828-35
pubmed: 25466545
Cancer Sci. 2013 Jun;104(6):663-71
pubmed: 23433103
Cancer Biol Ther. 2012 Oct;13(12):1221-8
pubmed: 22922788
Pigment Cell Melanoma Res. 2011 Feb;24(1):175-86
pubmed: 20955350
Nat Cell Biol. 2014 Jun;16(6):488-94
pubmed: 24875735
Mutat Res Rev Mutat Res. 2017 Apr - Jun;772:78-104
pubmed: 28528692
World J Surg Oncol. 2015 Oct 24;13:307
pubmed: 26499380
Lab Invest. 2016 Feb;96(2):197-205
pubmed: 26237271
Nature. 1997 Nov 6;390(6655):45-51
pubmed: 9363890
Dis Esophagus. 2016 Apr;29(3):207-14
pubmed: 25287007
Science. 2005 Sep 16;309(5742):1829-33
pubmed: 16123266
Pharm Res. 2012 Mar;29(3):643-50
pubmed: 21969054