Direct early growth response-1 knockdown decreases melanoma viability independent of mitogen-activated extracellular signal-related kinase inhibition.
Humans
Melanoma
/ drug therapy
Mitogens
Dimethyl Sulfoxide
Skin Neoplasms
/ drug therapy
Extracellular Signal-Regulated MAP Kinases
/ metabolism
Protein Kinase Inhibitors
/ pharmacology
Mitogen-Activated Protein Kinase Kinases
RNA, Small Interfering
Cell Line, Tumor
Proto-Oncogene Proteins B-raf
/ genetics
Journal
Melanoma research
ISSN: 1473-5636
Titre abrégé: Melanoma Res
Pays: England
ID NLM: 9109623
Informations de publication
Date de publication:
01 12 2023
01 12 2023
Historique:
pmc-release:
01
12
2024
medline:
30
10
2023
pubmed:
31
8
2023
entrez:
31
8
2023
Statut:
ppublish
Résumé
To investigate downstream molecular changes caused by mitogen-activated protein kinase (MEK) inhibitor treatment and further explore the impact of direct knockdown of early growth response-1 ( EGR1 ) in melanoma cell culture. RNA-sequencing (RNA-Seq) was performed to determine gene expression changes with MEK inhibitor treatment. Treatment with MEK inhibitor (trametinib) was then assessed in two cutaneous (MEL888, MEL624) and one conjunctival (YUARGE 13-3064) melanoma cell line. Direct knockdown of EGR1 was accomplished using lentiviral vectors containing shRNA. Cell viability was measured using PrestoBlueHS Cell Viability Reagent. Total RNA and protein were assessed by qPCR and SimpleWestern. RNA-Seq demonstrated a profound reduction in EGR1 with MEK inhibitor treatment, prompting further study of melanoma cell lines. Following trametinib treatment of melanoma cells, viability was reduced in both cutaneous (MEL888 26%, P < 0.01; MEL624 27%, P < 0.001) and conjunctival (YUARGE 13-3064 33%, P < 0.01) melanoma compared with DMSO control, with confirmed EGR1 knockdown to 0.04-, 0.01-, and 0.16-fold DMSO-treated levels (all P < 0.05) in MEL888, MEL624, and YUARGE 13-3064, respectively. Targeted EGR1 knockdown using shRNA reduced viability in both cutaneous (MEL624 78%, P = 0.05) and conjunctival melanoma (YUARGE-13-3064 67%, P = 0.02). RNA-Sequencing in MEK inhibitor-treated cells identified EGR1 as a candidate effector molecule of interest. In a malignant melanoma cell population, MEK inhibition reduced viability in both cutaneous and conjunctival melanoma with a profound downstream reduction in EGR1 expression. Targeted knockdown of EGR1 reduced both cutaneous and conjunctival melanoma cell viability independent of MEK inhibition, suggesting a key role for EGR1 in melanoma pathobiology.
Identifiants
pubmed: 37650708
doi: 10.1097/CMR.0000000000000921
pii: 00008390-990000000-00096
pmc: PMC10615778
mid: NIHMS1923785
doi:
Substances chimiques
Mitogens
0
Dimethyl Sulfoxide
YOW8V9698H
Extracellular Signal-Regulated MAP Kinases
EC 2.7.11.24
Protein Kinase Inhibitors
0
Mitogen-Activated Protein Kinase Kinases
EC 2.7.12.2
RNA, Small Interfering
0
Proto-Oncogene Proteins B-raf
EC 2.7.11.1
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
482-491Subventions
Organisme : NCATS NIH HHS
ID : KL2 TR002379
Pays : United States
Organisme : NCI NIH HHS
ID : P30 CA015083
Pays : United States
Informations de copyright
Copyright © 2023 Wolters Kluwer Health, Inc. All rights reserved.
Références
Cancer Res. 1998 Jun 1;58(11):2461-8
pubmed: 9622090
Mol Cell Biol. 1990 May;10(5):1931-9
pubmed: 2109185
BMC Bioinformatics. 2014 Jun 27;15:224
pubmed: 24972667
Cancer Cell. 2017 Aug 14;32(2):204-220.e15
pubmed: 28810145
Nat Commun. 2014 Dec 02;5:5694
pubmed: 25452114
Cells. 2020 Jan 13;9(1):
pubmed: 31941155
Biochim Biophys Acta. 2007 Jan;1775(1):21-62
pubmed: 16904831
Cell Discov. 2023 Mar 28;9(1):33
pubmed: 36977660
ISRN Mol Biol. 2012 Dec 24;2012:381428
pubmed: 27340590
Eur J Cancer. 2017 Mar;73:93-101
pubmed: 28162869
Biochim Biophys Acta. 2016 Apr;1863(4):770-84
pubmed: 26844774
Med Sci (Basel). 2021 Oct 20;9(4):
pubmed: 34698235
Invest Ophthalmol Vis Sci. 2015 Jul;56(8):4619-30
pubmed: 26200502
CA Cancer J Clin. 2007 Jan-Feb;57(1):43-66
pubmed: 17237035
Cancers (Basel). 2020 Sep 29;12(10):
pubmed: 33003483
Molecules. 2017 Sep 26;22(10):
pubmed: 28954413
iScience. 2021 Aug 30;24(9):103067
pubmed: 34541473
Eur J Cancer. 2018 Nov;103:41-51
pubmed: 30205280
J Clin Oncol. 2005 Mar 20;23(9):2078-93
pubmed: 15774796
BMC Syst Biol. 2014 Sep 11;8:100
pubmed: 25217033
Curr Cancer Drug Targets. 2004 Feb;4(1):43-52
pubmed: 14965266
Oncol Rep. 2021 Sep;46(3):
pubmed: 34296292
Genome Biol. 2014;15(12):550
pubmed: 25516281
Oncotarget. 2018 Apr 6;9(26):18084-18098
pubmed: 29719592
Genes Cancer. 2011 Sep;2(9):900-9
pubmed: 22593802
Cancer Res. 1995 Nov 1;55(21):5054-62
pubmed: 7585551
Clin Cancer Res. 2008 Jan 15;14(2):342-6
pubmed: 18223206
Cell. 2008 Jul 25;134(2):215-30
pubmed: 18662538
Cancer Res. 1994 Jan 15;54(2):575-81
pubmed: 8275496
BMC Bioinformatics. 2012 Jun 18;13:134
pubmed: 22708584
J Pathol. 1993 Nov;171(3):191-7
pubmed: 8277368
Cell Rep. 2019 May 21;27(8):2493-2507.e4
pubmed: 31116991
J Hematol Oncol. 2021 Apr 6;14(1):55
pubmed: 33823905
Life (Basel). 2020 Sep 16;10(9):
pubmed: 32948031
Cancer Res. 2020 Jun 15;80(12):2676-2688
pubmed: 32291316
Int J Mol Sci. 2020 Jun 27;21(13):
pubmed: 32605090
Oncogene. 2003 Jul 3;22(27):4194-204
pubmed: 12833142
Cancers (Basel). 2021 Aug 15;13(16):
pubmed: 34439267
Int J Cancer. 1997 Jul 3;72(1):102-9
pubmed: 9212230
Cytokine Growth Factor Rev. 2009 Aug;20(4):305-17
pubmed: 19656717
Int J Mol Sci. 2021 Jun 23;22(13):
pubmed: 34201614
Trends Cancer. 2020 Sep;6(9):797-810
pubmed: 32540454