Elucidating the role of EPPK1 in lung adenocarcinoma development.
CRISPR-Cas9
Epiplakin 1
Epithelial-to-mesenchymal transition
MYC/p53 pathway
Journal
BMC cancer
ISSN: 1471-2407
Titre abrégé: BMC Cancer
Pays: England
ID NLM: 100967800
Informations de publication
Date de publication:
10 Apr 2024
10 Apr 2024
Historique:
received:
27
11
2023
accepted:
26
03
2024
medline:
10
4
2024
pubmed:
10
4
2024
entrez:
9
4
2024
Statut:
epublish
Résumé
We recently found that epiplakin 1 (EPPK1) alterations were present in 12% of lung adenocarcinoma (LUAD) cases and were associated with a poor prognosis in early-stage LUAD when combined with other molecular alterations. This study aimed to identify a probable crucial role for EPPK1 in cancer development. EPPK1 mRNA and protein expression was analyzed with clinical variables. Normal bronchial epithelial cell lines were exposed to cigarette smoke for 16 weeks to determine whether EPPK1 protein expression was altered after exposure. Further, we used CRISPR-Cas9 to knock out (KO) EPPK1 in LUAD cell lines and observed how the cancer cells were altered functionally and genetically. EPPK1 protein expression was associated with smoking and poor prognosis in early-stage LUAD. Moreover, a consequential mesenchymal-to-epithelial transition was observed, subsequently resulting in diminished cell proliferation and invasion after EPPK1 KO. RNA sequencing revealed that EPPK1 KO induced downregulation of 11 oncogenes, 75 anti-apoptosis, and 22 angiogenesis genes while upregulating 8 tumor suppressors and 12 anti-cell growth genes. We also observed the downregulation of MYC and upregulation of p53 expression at both protein and RNA levels following EPPK1 KO. Gene ontology enrichment analysis of molecular functions highlighted the correlation of EPPK1 with the regulation of mesenchymal cell proliferation, mesenchymal differentiation, angiogenesis, and cell growth after EPPK1 KO. Our data suggest that EPPK1 is linked to smoking, epithelial to mesenchymal transition, and the regulation of cancer progression, indicating its potential as a therapeutic target for LUAD.
Sections du résumé
BACKGROUND
BACKGROUND
We recently found that epiplakin 1 (EPPK1) alterations were present in 12% of lung adenocarcinoma (LUAD) cases and were associated with a poor prognosis in early-stage LUAD when combined with other molecular alterations. This study aimed to identify a probable crucial role for EPPK1 in cancer development.
METHODS
METHODS
EPPK1 mRNA and protein expression was analyzed with clinical variables. Normal bronchial epithelial cell lines were exposed to cigarette smoke for 16 weeks to determine whether EPPK1 protein expression was altered after exposure. Further, we used CRISPR-Cas9 to knock out (KO) EPPK1 in LUAD cell lines and observed how the cancer cells were altered functionally and genetically.
RESULTS
RESULTS
EPPK1 protein expression was associated with smoking and poor prognosis in early-stage LUAD. Moreover, a consequential mesenchymal-to-epithelial transition was observed, subsequently resulting in diminished cell proliferation and invasion after EPPK1 KO. RNA sequencing revealed that EPPK1 KO induced downregulation of 11 oncogenes, 75 anti-apoptosis, and 22 angiogenesis genes while upregulating 8 tumor suppressors and 12 anti-cell growth genes. We also observed the downregulation of MYC and upregulation of p53 expression at both protein and RNA levels following EPPK1 KO. Gene ontology enrichment analysis of molecular functions highlighted the correlation of EPPK1 with the regulation of mesenchymal cell proliferation, mesenchymal differentiation, angiogenesis, and cell growth after EPPK1 KO.
CONCLUSIONS
CONCLUSIONS
Our data suggest that EPPK1 is linked to smoking, epithelial to mesenchymal transition, and the regulation of cancer progression, indicating its potential as a therapeutic target for LUAD.
Identifiants
pubmed: 38594604
doi: 10.1186/s12885-024-12185-x
pii: 10.1186/s12885-024-12185-x
doi:
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
441Subventions
Organisme : NIH HHS
ID : U01CA152662
Pays : United States
Organisme : NIH HHS
ID : U01CA152662
Pays : United States
Organisme : NIH HHS
ID : CA196405
Pays : United States
Informations de copyright
© 2024. The Author(s).
Références
Maemondo M, Inoue A, Kobayashi K, et al. Gefitinib or chemotherapy for non-small-cell lung cancer with mutated EGFR. N Engl J Med. 2010;362:2380–8. https://doi.org/10.1056/NEJMoa0909530 .
doi: 10.1056/NEJMoa0909530
pubmed: 20573926
Takeuchi K, Choi YL, Soda M, et al. Multiplex reverse transcription-PCR screening for EML4-ALK fusion transcripts. Clin Cancer Res. 2008;14:6618–24. https://doi.org/10.1158/1078-0432.CCR-08-1018 .
doi: 10.1158/1078-0432.CCR-08-1018
pubmed: 18927303
Shaw AT, Ou SH, Bang YJ, et al. Crizotinib in ROS1-rearranged non-small-cell lung cancer. N Engl J Med. 2014;371:1963–19671. https://doi.org/10.1056/NEJMoa1406766 .
doi: 10.1056/NEJMoa1406766
pubmed: 25264305
pmcid: 4264527
Drilon A, Oxnard GR, Tan DSW, et al. Efficacy of selpercatinib in RET fusion-positive non-small-cell lung cancer. N Engl J Med. 2020;383:813–24. https://doi.org/10.1056/NEJMoa2005653 .
doi: 10.1056/NEJMoa2005653
pubmed: 32846060
pmcid: 7506467
Planchard D, Smit EF, Groen HJM, et al. Dabrafenib plus trametinib in patients with previously untreated BRAFV600E-mutant metastatic non-small-cell lung cancer: an open-label, phase 2 trial. Lancet Oncol. 2017;18:1307–16. https://doi.org/10.1016/S1470-2045(17)30679-4 .
doi: 10.1016/S1470-2045(17)30679-4
pubmed: 28919011
Skoulidis F, Li BT, Dy GK, et al. Sotorasib for lung cancers with KRAS p. G12C mutation. N Engl J Med. 2021;384:2371–81. https://doi.org/10.1056/NEJMoa2103695 .
doi: 10.1056/NEJMoa2103695
pubmed: 34096690
pmcid: 9116274
Ji X, Qian J, Rahman SMJ, et al. xCT (SLC7A11)-mediated metabolic reprogramming promotes non-small cell lung cancer progression. Oncogene. 2018;37:5007–19. https://doi.org/10.1038/s41388-018-0307-z .
doi: 10.1038/s41388-018-0307-z
pubmed: 29789716
pmcid: 6127081
Jang SI, Kalinin A, Takahashi K, et al. Characterization of human epiplakin: RNAi-mediated epiplakin depletion leads to the disruption of keratin and vimentin IF networks. J Cell Sci. 2005;118:781–93. https://doi.org/10.1242/jcs.01647 .
doi: 10.1242/jcs.01647
pubmed: 15671067
Kokado M, Okada Y, Miyamoto T, et al. Effects of epiplakin-knockdown in cultured corneal epithelial cells. BMC Res Notes. 2016;9:278. https://doi.org/10.1186/s13104-016-2082-7 .
doi: 10.1186/s13104-016-2082-7
pubmed: 27206504
pmcid: 4873999
Shimada H, Nambu-Niibori A, Wilson-Morifuji M, et al. Epiplakin modifies the motility of the HeLa cells and accumulates at the outer surfaces of 3-D cell clusters. J Dermatol. 2013;40:249–58. https://doi.org/10.1111/1346-8138.12076 .
doi: 10.1111/1346-8138.12076
pubmed: 23398049
Qian J, Zhao S, Zou Y, et al. Genomic underpinnings of tumor behavior in in situ and early lung adenocarcinoma. Am J Respir Crit Care Med. 2020;201:697–706. https://doi.org/10.1164/rccm.201902-0294OC .
doi: 10.1164/rccm.201902-0294OC
pubmed: 31747302
pmcid: 7068818
Kokado M, Okada Y, Goto M, et al. Increased fragility, impaired differentiation, and acceleration of migration of corneal epithelium of epiplakin-null mice. Invest Ophthalmol Vis Sci. 2013;54:3780–9. https://doi.org/10.1167/iovs.12-11077 .
doi: 10.1167/iovs.12-11077
pubmed: 23599337
Borghaei H, Paz-Ares L, Horn L, et al. Nivolumab versus docetaxel in advanced nonsquamous non-small-cell lung cancer. N Engl J Med. 2015;373:1627–39. https://doi.org/10.1056/NEJMoa1507643 .
doi: 10.1056/NEJMoa1507643
pubmed: 26412456
pmcid: 5705936
Arimura K, Kondo M, Nagashima Y, et al. Comparison of tumor cell numbers and 22C3 PD-L1 expression between cryobiopsy and transbronchial biopsy with endobronchial ultrasonography-guide sheath for lung cancer. Respir Res. 2019;20:185. https://doi.org/10.1186/s12931-019-1162-3 .
doi: 10.1186/s12931-019-1162-3
pubmed: 31420048
pmcid: 6698028
Li T, Fan J, Wang B, et al. TIMER: A web server for comprehensive analysis of tumor-infiltrating immune cells. Cancer Res. 2017;77:e108-110. https://doi.org/10.1158/0008-5472.CAN-17-0307 .
doi: 10.1158/0008-5472.CAN-17-0307
pubmed: 29092952
pmcid: 6042652
Arimura K, Hiroshima K, Nagashima Y, et al. LAG3 is an independent prognostic biomarker and potential target for immune checkpoint inhibitors in malignant pleural mesothelioma: a retrospective study. BMC Cancer. 2023;23:1206. https://doi.org/10.1186/s12885-023-11636-1 .
doi: 10.1186/s12885-023-11636-1
pubmed: 38062416
pmcid: 10704683
Qian J, Zou Y, Rahman JS, et al. Synergy between phosphatidylinositol 3-kinase/Akt pathway and Bcl-xL in the control of apoptosis in adenocarcinoma cells of the lung. Mol Cancer Ther. 2009;8:101–9. https://doi.org/10.1158/1535-7163.MCT-08-0973 .
doi: 10.1158/1535-7163.MCT-08-0973
pubmed: 19139118
pmcid: 3110728
Dobin A, Davis CA, Schlesinger F, et al. STAR: ultrafast universal RNA-seq aligner. Bioinformatics. 2013;29:15–21. https://doi.org/10.1093/bioinformatics/bts635 .
doi: 10.1093/bioinformatics/bts635
pubmed: 23104886
Liao Y, Smyth GK, Shi W. featureCounts: an efficient general purpose program for assigning sequence reads to genomic features. Bioinformatics. 2014;30:923–30. https://doi.org/10.1093/bioinformatics/btt656 .
doi: 10.1093/bioinformatics/btt656
pubmed: 24227677
Love MI, Huber W, Anders S. Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2. Genome Biol. 2014;15:550. https://doi.org/10.1186/s13059-014-0550-8 .
doi: 10.1186/s13059-014-0550-8
pubmed: 25516281
pmcid: 4302049
Dongre A, Weinberg RA. New insights into the mechanisms of epithelial-mesenchymal transition and implications for cancer. Nat Rev Mol Cell Biol. 2019;20:69–84. https://doi.org/10.1038/s41580-018-0080-4 .
doi: 10.1038/s41580-018-0080-4
pubmed: 30459476
Hackshaw AK, Law MR, Wald NJ. The accumulated evidence on lung cancer and environmental tobacco smoke. BMJ. 1997;315:980–8. https://doi.org/10.1136/bmj.315.7114.980 .
doi: 10.1136/bmj.315.7114.980
pubmed: 9365295
pmcid: 2127653
Albino AP, Jorgensen ED, Rainey P, et al. gammaH2AX: a potential DNA damage response biomarker for assessing toxicological risk of tobacco products. Mutat Res. 2009;678:43–52. https://doi.org/10.1016/j.mrgentox.2009.06.009 .
doi: 10.1016/j.mrgentox.2009.06.009
pubmed: 19591958
de Bruin EC, McGranahan N, Mitter R, et al. Spatial and temporal diversity in genomic instability processes defines lung cancer evolution. Science. 2014;346:251–6. https://doi.org/10.1126/science.1253462 .
doi: 10.1126/science.1253462
pubmed: 25301630
pmcid: 4636050
Massion PP, Zou Y, Chen H, et al. Smoking-related genomic signatures in non-small cell lung cancer. Am J Respir Crit Care Med. 2008;178:1164–72. https://doi.org/10.1164/rccm.200801-142OC .
doi: 10.1164/rccm.200801-142OC
pubmed: 18776155
pmcid: 2720147
Shen T, Lu Y, Zhang Q. High squalene epoxidase in tumors predicts worse survival in patients with hepatocellular carcinoma: Integrated bioinformatic analysis on NAFLD and HCC. Cancer Control. 2020;27:1073274820914663. https://doi.org/10.1177/1073274820914663 .
doi: 10.1177/1073274820914663
pubmed: 32216563
pmcid: 7137641
Qiao Z, Dai C, Wang Z, et al. Epiplakin1 promotes the progression of esophageal squamous cell carcinoma by activating the PI3K-AKT signaling pathway. Thorac Cancer. 2022;13:1117–25. https://doi.org/10.1111/1759-7714.14366 .
doi: 10.1111/1759-7714.14366
pubmed: 35238170
pmcid: 9013648
Yoshida T, Shiraki N, Baba H, et al. Expression patterns of epiplakin1 in pancreas, pancreatic cancer and regenerating pancreas. Genes Cells. 2008;13:667–78. https://doi.org/10.1111/j.1365-2443.2008.01196.x .
doi: 10.1111/j.1365-2443.2008.01196.x
pubmed: 18498355
Guo X, Hao Y, Kamilijiang M, et al. Potential predictive plasma biomarkers for cervical cancer by 2D-DIGE proteomics and ingenuity pathway analysis. Tumour Biol. 2015;36:1711–1120. https://doi.org/10.1007/s13277-014-2772-5 .
doi: 10.1007/s13277-014-2772-5
pubmed: 25427637
Pattabiraman DR, Bierie B, Kober KI, et al. Activation of PKA leads to mesenchymal-to-epithelial transition and loss of tumor-initiating ability. Science. 2016;351:aad3680. https://doi.org/10.1126/science.aad3680 .
doi: 10.1126/science.aad3680
pubmed: 26941323
pmcid: 5131720
Olivero CE, Martínez-Terroba E, Zimmer J, et al. p53 activates the long noncoding RNA Pvt1b to inhibit myc and suppress tumorigenesis. Mol Cell. 2020;77:761-774.e8. https://doi.org/10.1016/j.molcel.2019.12.014 .
doi: 10.1016/j.molcel.2019.12.014
pubmed: 31973890
pmcid: 7184554
Ho JS, Ma W, Mao DY, Benchimol S. p53-Dependent transcriptional repression of c-myc is required for G1 cell cycle arrest. Mol Cell Biol. 2005;25:7423–31. https://doi.org/10.1128/MCB.25.17.7423-7431.2005 .
doi: 10.1128/MCB.25.17.7423-7431.2005
pubmed: 16107691
pmcid: 1190302
Felsher DW, Zetterberg A, Zhu J, et al. Overexpression of MYC causes p53-dependent G2 arrest of normal fibroblasts. Proc Natl Acad Sci U S A. 2000;97:10544–8. https://doi.org/10.1073/pnas.190327097 .
doi: 10.1073/pnas.190327097
pubmed: 10962037
pmcid: 27061