Opposing Functions of Maspin Are Regulated by Its Subcellular Localization in Lung Squamous Cell Carcinoma Cells
Abstract
:Simple Summary
Abstract
1. Introduction
2. Materials and Methods
2.1. Cell Culture
2.2. Real-Time Quantitative Polymerase Chain Reaction (RT-qPCR)
2.3. Western Blotting
2.4. Immunofluorescence
2.5. siRNA Transfection
2.6. Establishment of RERF-LC-AI and LK-2 Cell Lines with High Expression of Maspin
2.7. Cell Invasion Assay
2.8. Cell Migration Assay
2.9. Transcriptome Analysis (mRNA-Seq)
2.10. Statistical Analysis
3. Results
3.1. Expression Status and Subcellular Localization of Maspin in LUSC Cell Lines
3.2. Promotion of Invasive Capability by panMaspin Knockdown in LUSC Cell Line
3.3. Maspin Has Subcellular Localization-Dependent Opposing Effects on Cell Invasion and Migration
3.4. Maspin Re-Expression-Induced Alterations in Gene Expression in LUSC Cell Lines
3.5. PYK2 and SRC Activation in LC-AI-Maspin Cells
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
- Bray, F.; Laversanne, M.; Sung, H.; Ferlay, J.; Siegel, R.L.; Soerjomataram, I.; Jemal, A. Global Cancer Statistics 2022: GLOBOCAN Estimates of Incidence and Mortality Worldwide for 36 Cancers in 185 Countries. CA A Cancer J. Clin. 2024, 74, 229–263. [Google Scholar] [CrossRef] [PubMed]
- Barta, J.A.; Powell, C.A.; Wisnivesky, J.P. Global Epidemiology of Lung Cancer. Ann. Glob. Health 2019, 85, 8. [Google Scholar] [CrossRef] [PubMed]
- Desai, A.; Adjei, A.A. FGFR Signaling as a Target for Lung Cancer Therapy. J. Thorac. Oncol. 2016, 11, 9–20. [Google Scholar] [CrossRef] [PubMed]
- Dotto, G.P.; Rustgi, A.K. Squamous Cell Cancers: A Unified Perspective on Biology and Genetics. Cancer Cell 2016, 29, 622–637. [Google Scholar] [CrossRef] [PubMed]
- Lau, S.C.M.; Pan, Y.; Velcheti, V.; Wong, K.K. Squamous Cell Lung Cancer: Current Landscape and Future Therapeutic Options. Cancer Cell 2022, 40, 1279–1293. [Google Scholar] [CrossRef]
- Felip, E.; Altorki, N.; Zhou, C.; Csőszi, T.; Vynnychenko, I.; Goloborodko, O.; Luft, A.; Akopov, A.; Martinez-Marti, A.; Kenmotsu, H.; et al. Adjuvant Atezolizumab after Adjuvant Chemotherapy in Resected Stage IB–IIIA Non-Small-Cell Lung Cancer (IMpower010): A Randomised, Multicentre, Open-Label, Phase 3 Trial. Lancet 2021, 398, 1344–1357. [Google Scholar] [CrossRef]
- Paz-Ares, L.; Vicente, D.; Tafreshi, A.; Robinson, A.; Soto Parra, H.; Mazières, J.; Hermes, B.; Cicin, I.; Medgyasszay, B.; Rodríguez-Cid, J.; et al. A Randomized, Placebo-Controlled Trial of Pembrolizumab Plus Chemotherapy in Patients With Metastatic Squamous NSCLC: Protocol-Specified Final Analysis of KEYNOTE-407. J. Thorac. Oncol. 2020, 15, 1657–1669. [Google Scholar] [CrossRef]
- Zou, Z.; Anisowicz, A.; Hendrix, M.J.C.; Thor, A.; Neveu, M.; Sheng, S.; Rafidi, K.; Seftor, E.; Sager, R. Maspin, a Serpin with Tumor-Suppressing Activity in Human Mammary Epithelial Cells. Science 1994, 263, 526–529. [Google Scholar] [CrossRef]
- Wang, M.-C.; Yang, Y.-M.; Li, X.-H.; Dong, F.; Li, Y. Maspin Expression and Its Clinicopathological Significance in Tumorigenesis and Progression of Gastric Cancer. World J. Gastroenterol. 2004, 10, 634–637. [Google Scholar] [CrossRef]
- Dabiri, S.; Aghtaei, M.M.; Shahryari, J.; Meymandi, S.; Amirpour-Rostami, S.; Foutohi-Ardekani, R. Maspin Gene Expression in Invasive Ductal Carcinoma of Breast. Iran. J. Pathol. 2016, 11, 104–111. [Google Scholar]
- Zou, Z.; Gao, C.; Nagaich, A.K.; Connell, T.; Saito, S.; Moul, J.W.; Seth, P.; Appella, E.; Srivastava, S. P53 Regulates the Expression of the Tumor Suppressor Gene Maspin. J. Biol. Chem. 2000, 275, 6051–6054. [Google Scholar] [CrossRef]
- Lin, Y.-H.; Tsui, K.-H.; Chang, K.-S.; Hou, C.-P.; Feng, T.-H.; Juang, H.-H. Maspin Is a PTEN-Upregulated and P53-Upregulated Tumor Suppressor Gene and Acts as an HDAC1 Inhibitor in Human Bladder Cancer. Cancers 2019, 12, 10. [Google Scholar] [CrossRef] [PubMed]
- Latha, K.; Zhang, W.; Cella, N.; Shi, H.Y.; Zhang, M. Maspin Mediates Increased Tumor Cell Apoptosis upon Induction of the Mitochondrial Permeability Transition. Mol. Cell Biol. 2005, 25, 1737–1748. [Google Scholar] [CrossRef] [PubMed]
- Bodenstine, T.M.; Seftor, R.E.B.; Khalkhali-Ellis, Z.; Seftor, E.A.; Pemberton, P.A.; Hendrix, M.J.C. Maspin: Molecular Mechanisms and Therapeutic Implications. Cancer Metastasis Rev. 2012, 31, 529–551. [Google Scholar] [CrossRef] [PubMed]
- Tang, S.; Ling, Z.; Jiang, J.; Gu, X.; Leng, Y.; Wei, C.; Cheng, H.; Li, X. Integrating the Tumor-Suppressive Activity of Maspin with P53 in Retuning the Epithelial Homeostasis: A Working Hypothesis and Applicable Prospects. Front. Oncol. 2022, 12, 1037794. [Google Scholar] [CrossRef] [PubMed]
- Matsuoka, Y.; Takagi, Y.; Nosaka, K.; Sakabe, T.; Haruki, T.; Araki, K.; Taniguchi, Y.; Shiomi, T.; Nakamura, H.; Umekita, Y. Cytoplasmic Expression of Maspin Predicts Unfavourable Prognosis in Patients with Squamous Cell Carcinoma of the Lung. Histopathology 2016, 69, 114–120. [Google Scholar] [CrossRef]
- Takagi, Y.; Matsuoka, Y.; Shiomi, T.; Nosaka, K.; Takeda, C.; Haruki, T.; Araki, K.; Taniguchi, Y.; Nakamura, H.; Umekita, Y. Cytoplasmic Maspin Expression Is a Predictor of Poor Prognosis in Patients with Lung Adenocarcinoma Measuring <3 Cm. Histopathology 2015, 66, 732–739. [Google Scholar] [CrossRef]
- Umekita, Y.; Ohi, Y.; Sagara, Y.; Yoshida, H. Expression of Maspin Predicts Poor Prognosis in Breast-Cancer Patients. Int. J. Cancer 2002, 100, 452–455. [Google Scholar] [CrossRef]
- Sakabe, T.; Wakahara, M.; Shiota, G.; Umekita, Y. Role of Cytoplasmic Localization of Maspin in Promoting Cell Invasion in Breast Cancer with Aggressive Phenotype. Sci. Rep. 2021, 11, 11321. [Google Scholar] [CrossRef]
- Goulet, B.; Kennette, W.; Ablack, A.; Postenka, C.O.; Hague, M.N.; Mymryk, J.S.; Tuck, A.B.; Giguère, V.; Chambers, A.F.; Lewis, J.D. Nuclear Localization of Maspin Is Essential for Its Inhibition of Tumor Growth and Metastasis. Lab. Investig. 2011, 91, 1181–1187. [Google Scholar] [CrossRef]
- Capes-Davis, A.; Theodosopoulos, G.; Atkin, I.; Drexler, H.G.; Kohara, A.; MacLeod, R.A.F.; Masters, J.R.; Nakamura, Y.; Reid, Y.A.; Reddel, R.R.; et al. Check Your Cultures! A List of Cross-Contaminated or Misidentified Cell Lines. Int. J. Cancer 2010, 127, 1–8. [Google Scholar] [CrossRef] [PubMed]
- Suarez-Arnedo, A.; Torres Figueroa, F.; Clavijo, C.; Arbeláez, P.; Cruz, J.C.; Muñoz-Camargo, C. An Image J Plugin for the High Throughput Image Analysis of in Vitro Scratch Wound Healing Assays. PLoS ONE 2020, 15, e0232565. [Google Scholar] [CrossRef] [PubMed]
- Matsushige, T.; Sakabe, T.; Umekita, Y. Investigation of the Subcellular Localization-Dependent Anti- or Pro-Tumor Functions of Maspin in Human Lung Adenocarcinoma Cell Line. Yonago Acta Med. 2022, 65, 44–52. [Google Scholar] [CrossRef] [PubMed]
- Shen, T.; Guo, Q. Role of Pyk2 in Human Cancers. Med. Sci. Monit. 2018, 24, 8172–8182. [Google Scholar] [CrossRef]
- Li, X.; Dy, R.C.; Cance, W.G.; Graves, L.M.; Earp, H.S. Interactions between Two Cytoskeleton-Associated Tyrosine Kinases: Calcium-Dependent Tyrosine Kinase and Focal Adhesion Tyrosine Kinase. J. Biol. Chem. 1999, 274, 8917–8924. [Google Scholar] [CrossRef]
- Woenckhaus, M.; Bubendorf, L.; Dalquen, P.; Foerster, J.; Blaszyk, H.; Mirlacher, M.; Soler, M.; Dietmaier, W.; Sauter, G.; Hartmann, A.; et al. Nuclear and Cytoplasmic Maspin Expression in Primary Non-Small Cell Lung Cancer. J. Clin. Pathol. 2007, 60, 483–486. [Google Scholar] [CrossRef]
- Nakagawa, M.; Katakura, H.; Adachi, M.; Takenaka, K.; Yanagihara, K.; Otake, Y.; Wada, H.; Tanaka, F. Maspin Expression and Its Clinical Significance in Non-Small Cell Lung Cancer. Ann. Surg. Oncol. 2006, 13, 1517–1523. [Google Scholar] [CrossRef]
- Takanami, I.; Abiko, T.; Koizumi, S. Expression of Maspin in Non–Small-Cell Lung Cancer: Correlation with Clinical Features. Clinical Lung Cancer 2008, 9, 361–366. [Google Scholar] [CrossRef]
- Lam, Y.W.; Yuan, Y.; Isaac, J.; Babu, C.V.S.; Meller, J.; Ho, S.-M. Comprehensive Identification and Modified-Site Mapping of S-Nitrosylated Targets in Prostate Epithelial Cells. PLoS ONE 2010, 5, e9075. [Google Scholar] [CrossRef]
- Zhao, S.; Xu, W.; Jiang, W.; Yu, W.; Lin, Y.; Zhang, T.; Yao, J.; Zhou, L.; Zeng, Y.; Li, H.; et al. Regulation of Cellular Metabolism by Protein Lysine Acetylation. Science 2010, 327, 1000–1004. [Google Scholar] [CrossRef]
- Odero-Marah, V.A.; Khalkhali-Ellis, Z.; Schneider, G.B.; Seftor, E.A.; Seftor, R.E.B.; Koland, J.G.; Hendrix, M.J.C. Tyrosine Phosphorylation of Maspin in Normal Mammary Epithelia and Breast Cancer Cells. Biochem. Biophys. Res. Commun. 2002, 295, 800–805. [Google Scholar] [CrossRef]
- Tamazato Longhi, M.; Cella, N. Tyrosine Phosphorylation Plays a Role in Increasing Maspin Protein Levels and Its Cytoplasmic Accumulation. FEBS Open Bio 2012, 2, 93–97. [Google Scholar] [CrossRef] [PubMed]
- Tamazato Longhi, M.; Magalhães, M.; Reina, J.; Morais Freitas, V.; Cella, N. EGFR Signaling Regulates Maspin/SerpinB5 Phosphorylation and Nuclear Localization in Mammary Epithelial Cells. PLoS ONE 2016, 11, e0159856. [Google Scholar] [CrossRef] [PubMed]
- Longhi, M.T.; Silva, L.E.; Pereira, M.; Magalhães, M.; Reina, J.; Vitorino, F.N.L.; Gumbiner, B.M.; da Cunha, J.P.C.; Cella, N. PI3K-AKT, JAK2-STAT3 Pathways and Cell-Cell Contact Regulate Maspin Subcellular Localization. Cell Commun. Signal 2021, 19, 86. [Google Scholar] [CrossRef] [PubMed]
- Tanaka, K.; Kumano, K.; Ueno, H. Intracellular Signals of Lung Cancer Cells as Possible Therapeutic Targets. Cancer Sci. 2015, 106, 489–496. [Google Scholar] [CrossRef]
- Zhang, L.; Bing, S.; Dong, M.; Lu, X.; Xiong, Y. Targeting Ion Channels for the Treatment of Lung Cancer. Biochim. Biophys. Acta Rev. Cancer 2021, 1876, 188629. [Google Scholar] [CrossRef]
- Alam, M.; Hasan, G.M.; Eldin, S.M.; Adnan, M.; Riaz, M.B.; Islam, A.; Khan, I.; Hassan, M.I. Investigating Regulated Signaling Pathways in Therapeutic Targeting of Non-Small Cell Lung Carcinoma. Biomed. Pharmacother. 2023, 161, 114452. [Google Scholar] [CrossRef]
- Iiizumi, M.; Bandyopadhyay, S.; Pai, S.K.; Watabe, M.; Hirota, S.; Hosobe, S.; Tsukada, T.; Miura, K.; Saito, K.; Furuta, E.; et al. RhoC Promotes Metastasis via Activation of the Pyk2 Pathway in Prostate Cancer. Cancer Res. 2008, 68, 7613–7620. [Google Scholar] [CrossRef] [PubMed]
- Wehrle-Haller, B.; Imhof, B. The Inner Lives of Focal Adhesions. Trends Cell Biol. 2002, 12, 382–389. [Google Scholar] [CrossRef] [PubMed]
- Ridley, A.J.; Schwartz, M.A.; Burridge, K.; Firtel, R.A.; Ginsberg, M.H.; Borisy, G.; Parsons, J.T.; Horwitz, A.R. Cell Migration: Integrating Signals from Front to Back. Science 2003, 302, 1704–1709. [Google Scholar] [CrossRef]
- Hashido, M.; Hayashi, K.; Hirose, K.; Iino, M. Ca2+ Lightning Conveys Cell-Cell Contact Information inside the Cells. EMBO Rep. 2006, 7, 1117–1123. [Google Scholar] [CrossRef] [PubMed]
- Avizienyte, E.; Frame, M.C. Src and FAK Signalling Controls Adhesion Fate and the Epithelial-to-Mesenchymal Transition. Curr. Opin. Cell Biol. 2005, 17, 542–547. [Google Scholar] [CrossRef] [PubMed]
- Sun, C.K.; Ng, K.T.; Lim, Z.X.; Cheng, Q.; Lo, C.M.; Poon, R.T.; Man, K.; Wong, N.; Fan, S.T. Proline-Rich Tyrosine Kinase 2 (Pyk2) Promotes Cell Motility of Hepatocellular Carcinoma through Induction of Epithelial to Mesenchymal Transition. PLoS ONE 2011, 6, e18878. [Google Scholar] [CrossRef] [PubMed]
- Liu, B.-X.; Xie, Y.; Zhang, J.; Zeng, S.; Li, J.; Tao, Q.; Yang, J.; Chen, Y.; Zeng, C. SERPINB5 Promotes Colorectal Cancer Invasion and Migration by Promoting EMT and Angiogenesis via the TNF-α/NF-κB Pathway. Int. Immunopharmacol. 2024, 131, 111759. [Google Scholar] [CrossRef]
- Wendt, M.K.; Schiemann, B.J.; Parvani, J.G.; Lee, Y.-H.; Kang, Y.; Schiemann, W.P. TGF-β Stimulates Pyk2 Expression as Part of an Epithelial-Mesenchymal Transition Program Required for Metastatic Outgrowth of Breast Cancer. Oncogene 2013, 32, 2005–2015. [Google Scholar] [CrossRef]
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Matsushige, T.; Sakabe, T.; Mochida, H.; Umekita, Y. Opposing Functions of Maspin Are Regulated by Its Subcellular Localization in Lung Squamous Cell Carcinoma Cells. Cancers 2024, 16, 3009. https://doi.org/10.3390/cancers16173009
Matsushige T, Sakabe T, Mochida H, Umekita Y. Opposing Functions of Maspin Are Regulated by Its Subcellular Localization in Lung Squamous Cell Carcinoma Cells. Cancers. 2024; 16(17):3009. https://doi.org/10.3390/cancers16173009
Chicago/Turabian StyleMatsushige, Takahiro, Tomohiko Sakabe, Hirotoshi Mochida, and Yoshihisa Umekita. 2024. "Opposing Functions of Maspin Are Regulated by Its Subcellular Localization in Lung Squamous Cell Carcinoma Cells" Cancers 16, no. 17: 3009. https://doi.org/10.3390/cancers16173009