MicroRNA-1 attenuates the growth and metastasis of small cell lung cancer through CXCR4/FOXM1/RRM2 axis.
CXCR4
FOXM1
Neuroendocrine carcinoma
RRM2
Small cell lung cancer
microRNAs
Journal
Molecular cancer
ISSN: 1476-4598
Titre abrégé: Mol Cancer
Pays: England
ID NLM: 101147698
Informations de publication
Date de publication:
04 01 2023
04 01 2023
Historique:
received:
08
04
2022
accepted:
06
12
2022
entrez:
3
1
2023
pubmed:
4
1
2023
medline:
6
1
2023
Statut:
epublish
Résumé
Small cell lung cancer (SCLC) is an aggressive lung cancer subtype that is associated with high recurrence and poor prognosis. Due to lack of potential drug targets, SCLC patients have few therapeutic options. MicroRNAs (miRNAs) provide an interesting repertoire of therapeutic molecules; however, the identification of miRNAs regulating SCLC growth and metastasis and their precise regulatory mechanisms are not well understood. To identify novel miRNAs regulating SCLC, we performed miRNA-sequencing from donor/patient serum samples and analyzed the bulk RNA-sequencing data from the tumors of SCLC patients. Further, we developed a nanotechnology-based, highly sensitive method to detect microRNA-1 (miR-1, identified miRNA) in patient serum samples and SCLC cell lines. To assess the therapeutic potential of miR-1, we developed various in vitro models, including miR-1 sponge (miR-1Zip) and DOX-On-miR-1 (Tet-ON) inducible stable overexpression systems. Mouse models derived from intracardiac injection of SCLC cells (miR-1Zip and DOX-On-miR-1) were established to delineate the role of miR-1 in SCLC metastasis. In situ hybridization and immunohistochemistry were used to analyze the expression of miR-1 and target proteins (mouse and human tumor specimens), respectively. Dual-luciferase assay was used to validate the target of miR-1, and chromatin immunoprecipitation assay was used to investigate the protein-gene interactions. A consistent downregulation of miR-1 was observed in tumor tissues and serum samples of SCLC patients compared to their matched normal controls, and these results were recapitulated in SCLC cell lines. Gain of function studies of miR-1 in SCLC cell lines showed decreased cell growth and oncogenic signaling, whereas loss of function studies of miR-1 rescued this effect. Intracardiac injection of gain of function of miR-1 SCLC cell lines in the mouse models showed a decrease in distant organ metastasis, whereas loss of function of miR-1 potentiated growth and metastasis. Mechanistic studies revealed that CXCR4 is a direct target of miR-1 in SCLC. Using unbiased transcriptomic analysis, we identified CXCR4/FOXM1/RRM2 as a unique axis that regulates SCLC growth and metastasis. Our results further showed that FOXM1 directly binds to the RRM2 promoter and regulates its activity in SCLC. Our findings revealed that miR-1 is a critical regulator for decreasing SCLC growth and metastasis. It targets the CXCR4/FOXM1/RRM2 axis and has a high potential for the development of novel SCLC therapies. MicroRNA-1 (miR-1) downregulation in the tumor tissues and serum samples of SCLC patients is an important hallmark of tumor growth and metastasis. The introduction of miR-1 in SCLC cell lines decreases cell growth and metastasis. Mechanistically, miR-1 directly targets CXCR4, which further prevents FOXM1 binding to the RRM2 promoter and decreases SCLC growth and metastasis.
Sections du résumé
BACKGROUND
Small cell lung cancer (SCLC) is an aggressive lung cancer subtype that is associated with high recurrence and poor prognosis. Due to lack of potential drug targets, SCLC patients have few therapeutic options. MicroRNAs (miRNAs) provide an interesting repertoire of therapeutic molecules; however, the identification of miRNAs regulating SCLC growth and metastasis and their precise regulatory mechanisms are not well understood.
METHODS
To identify novel miRNAs regulating SCLC, we performed miRNA-sequencing from donor/patient serum samples and analyzed the bulk RNA-sequencing data from the tumors of SCLC patients. Further, we developed a nanotechnology-based, highly sensitive method to detect microRNA-1 (miR-1, identified miRNA) in patient serum samples and SCLC cell lines. To assess the therapeutic potential of miR-1, we developed various in vitro models, including miR-1 sponge (miR-1Zip) and DOX-On-miR-1 (Tet-ON) inducible stable overexpression systems. Mouse models derived from intracardiac injection of SCLC cells (miR-1Zip and DOX-On-miR-1) were established to delineate the role of miR-1 in SCLC metastasis. In situ hybridization and immunohistochemistry were used to analyze the expression of miR-1 and target proteins (mouse and human tumor specimens), respectively. Dual-luciferase assay was used to validate the target of miR-1, and chromatin immunoprecipitation assay was used to investigate the protein-gene interactions.
RESULTS
A consistent downregulation of miR-1 was observed in tumor tissues and serum samples of SCLC patients compared to their matched normal controls, and these results were recapitulated in SCLC cell lines. Gain of function studies of miR-1 in SCLC cell lines showed decreased cell growth and oncogenic signaling, whereas loss of function studies of miR-1 rescued this effect. Intracardiac injection of gain of function of miR-1 SCLC cell lines in the mouse models showed a decrease in distant organ metastasis, whereas loss of function of miR-1 potentiated growth and metastasis. Mechanistic studies revealed that CXCR4 is a direct target of miR-1 in SCLC. Using unbiased transcriptomic analysis, we identified CXCR4/FOXM1/RRM2 as a unique axis that regulates SCLC growth and metastasis. Our results further showed that FOXM1 directly binds to the RRM2 promoter and regulates its activity in SCLC.
CONCLUSIONS
Our findings revealed that miR-1 is a critical regulator for decreasing SCLC growth and metastasis. It targets the CXCR4/FOXM1/RRM2 axis and has a high potential for the development of novel SCLC therapies. MicroRNA-1 (miR-1) downregulation in the tumor tissues and serum samples of SCLC patients is an important hallmark of tumor growth and metastasis. The introduction of miR-1 in SCLC cell lines decreases cell growth and metastasis. Mechanistically, miR-1 directly targets CXCR4, which further prevents FOXM1 binding to the RRM2 promoter and decreases SCLC growth and metastasis.
Identifiants
pubmed: 36597126
doi: 10.1186/s12943-022-01695-6
pii: 10.1186/s12943-022-01695-6
pmc: PMC9811802
doi:
Substances chimiques
MicroRNAs
0
FOXM1 protein, human
0
Forkhead Box Protein M1
0
CXCR4 protein, human
0
Receptors, CXCR4
0
MIRN1 microRNA, human
0
Mirn1 microRNA, mouse
0
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, U.S. Gov't, Non-P.H.S.
Langues
eng
Sous-ensembles de citation
IM
Pagination
1Subventions
Organisme : NIH HHS
ID : R01CA218545
Pays : United States
Organisme : NCI NIH HHS
ID : R01 CA247471
Pays : United States
Organisme : NCI NIH HHS
ID : R01 CA195586
Pays : United States
Organisme : NIH HHS
ID : R01CA247471
Pays : United States
Organisme : NCI NIH HHS
ID : P30 CA033572
Pays : United States
Organisme : NCI NIH HHS
ID : P01 CA217798
Pays : United States
Organisme : NCI NIH HHS
ID : R01 CA218545
Pays : United States
Organisme : NCI NIH HHS
ID : R01 CA241752
Pays : United States
Informations de copyright
© 2023. The Author(s).
Références
EMBO Mol Med. 2021 Jan 11;13(1):e13122
pubmed: 33296145
Brain Pathol. 2020 Jul;30(4):732-745
pubmed: 32145124
Nat Commun. 2016 Nov 15;7:13398
pubmed: 27845331
Nat Rev Cancer. 2019 May;19(5):289-297
pubmed: 30926931
Gastroenterology. 2020 Nov;159(5):1898-1915.e6
pubmed: 32781084
Sci Transl Med. 2019 Nov 6;11(517):
pubmed: 31694929
Cell. 2009 Jan 23;136(2):215-33
pubmed: 19167326
Nat Commun. 2014 Nov 12;5:5165
pubmed: 25387393
FEBS Lett. 2012 Oct 19;586(20):3639-44
pubmed: 22992418
Nat Genet. 2005 May;37(5):495-500
pubmed: 15806104
Nanomedicine. 2022 Jul;43:102566
pubmed: 35569810
Lancet. 2015 Jan 3;385(9962):9-10
pubmed: 25230596
Cancer Discov. 2018 Nov;8(11):1422-1437
pubmed: 30181244
Genes Dev. 2009 Sep 15;23(18):2152-65
pubmed: 19759263
Cancer Cell. 2021 Nov 8;39(11):1479-1496.e18
pubmed: 34653364
Nucleic Acids Res. 2019 Jan 8;47(D1):D155-D162
pubmed: 30423142
FEBS Lett. 2010 Aug 20;584(16):3592-600
pubmed: 20655308
Mol Cancer. 2022 Jan 24;21(1):29
pubmed: 35073911
Semin Cell Dev Biol. 2022 Apr;124:114-126
pubmed: 34034986
Clin Cancer Res. 2019 Jul 15;25(14):4480-4492
pubmed: 30996073
Nat Genet. 2012 Oct;44(10):1111-6
pubmed: 22941189
Nat Immunol. 2019 Dec;20(12):1621-1630
pubmed: 31740800
Mol Cancer Ther. 2011 Jun;10(6):1046-58
pubmed: 21518729
Nat Cell Biol. 2005 Feb;7(2):126-36
pubmed: 15654331
Nat Rev Genet. 2013 Aug;14(8):535-48
pubmed: 23817310
Proc Natl Acad Sci U S A. 2011 Mar 22;108(12):5003-8
pubmed: 21383194
Nucleic Acids Res. 1997 May 1;25(9):1715-9
pubmed: 9108152
Int Cancer Conf J. 2019 Mar 7;8(3):109-113
pubmed: 31218185
Anal Chem. 2015 Nov 3;87(21):10822-9
pubmed: 26451797
Lung Cancer. 2017 Mar;105:7-13
pubmed: 28236984
Nature. 2012 Sep 6;489(7414):101-8
pubmed: 22955620
Bone Res. 2022 Jan 20;10(1):6
pubmed: 35058441
Cancer Discov. 2021 Aug;11(8):1952-1969
pubmed: 33707236
Proc Natl Acad Sci U S A. 2020 Sep 29;117(39):24213-24223
pubmed: 32929008
Cancer Res. 2002 Nov 1;62(21):6304-11
pubmed: 12414661
Cancer. 2015 Mar 1;121(5):664-72
pubmed: 25336398
Dev Cell. 2006 Oct;11(4):441-50
pubmed: 17011485
Adv Sci (Weinh). 2021 Sep;8(18):e2100881
pubmed: 34319001
Oncogenesis. 2016 Apr 18;5:e219
pubmed: 27089142
Mol Cancer. 2021 Dec 20;20(1):170
pubmed: 34930277
Lancet. 2011 Nov 12;378(9804):1741-55
pubmed: 21565397
Semin Cancer Biol. 2022 Dec;87:117-126
pubmed: 36371025
Nat Rev Dis Primers. 2021 Jan 14;7(1):3
pubmed: 33446664
Cancer Res. 2017 Jul 15;77(14):3931-3941
pubmed: 28487384
Sci Signal. 2019 Feb 05;12(567):
pubmed: 30723171
Nat Genet. 2012 Oct;44(10):1104-10
pubmed: 22941188
Oncogene. 2009 Dec 3;28(48):4295-305
pubmed: 19749794
Oncogene. 2021 Jul;40(30):4847-4858
pubmed: 34155349
Cancer Cell. 2015 Jul 13;28(1):57-69
pubmed: 26175415
J Thorac Oncol. 2020 Apr;15(4):618-627
pubmed: 31870883
Cancer Discov. 2019 Feb;9(2):230-247
pubmed: 30373918
Semin Cancer Biol. 2022 Aug;83:57-76
pubmed: 33220460
Invest New Drugs. 2017 Jun;35(3):334-344
pubmed: 28299514
Cancer Cell. 2017 Feb 13;31(2):286-299
pubmed: 28196596
Eur Rev Med Pharmacol Sci. 2019 Oct;23(20):8870-8877
pubmed: 31696489
Nature. 2005 Jul 14;436(7048):214-20
pubmed: 15951802
Nature. 2004 Sep 16;431(7006):350-5
pubmed: 15372042
Nature. 2015 Aug 6;524(7563):47-53
pubmed: 26168399
Oncogene. 2003 Nov 6;22(50):8093-101
pubmed: 14603250
Cancer Res. 2014 Feb 1;74(3):738-750
pubmed: 24310399
Oncogene. 2005 Jun 23;24(27):4462-71
pubmed: 15806155
Cancer Res. 2008 Jul 1;68(13):5049-58
pubmed: 18593903
J Med Chem. 1995 Sep 15;38(19):3865-73
pubmed: 7562918
J Clin Oncol. 2010 Apr 1;28(10):1721-6
pubmed: 20194856
Nat Genet. 2006 Feb;38(2):228-33
pubmed: 16380711
Cell. 2012 May 25;149(5):1023-34
pubmed: 22632967
Mol Cancer. 2021 Mar 19;20(1):54
pubmed: 33740988