Macrophages Promote Ovarian Cancer-Mesothelial Cell Adhesion by Upregulation of ITGA2 and VEGFC in Mesothelial Cells
Abstract
:1. Introduction
2. Materials and Methods
2.1. Materials
2.2. Cell Culture and Conditioned Medium
2.3. Adhesion Assay
2.4. RNA Isolation and Real-Time RT-PCR
2.5. Gene Knockdown Using Small Interfering RNA
2.6. mRNA Sequencing
2.7. Cytokine Antibody Arrays
2.8. Statistical Analysis
3. Results
3.1. Ovarian Cancer Cells Showed an Increased Adhesion to Macrophage-Stimulated Mesothelial Cells
3.2. Enhanced Expression of Adhesion-Related Genes in Macrophage-Stimulated Mesothelial Cells
3.3. Increased Expression of ITGA2 and VEGFC in Macrophage-Stimulated Mesothelial Cells Promotes Ovarian Cancer-Mesothelial Cell Adhesion
3.4. JNK and Akt Pathways in Macrophage-Stimulated Mesothelial Cells Were Associated with Enhanced Ovarian Cancer-Mesothelial Cell Adhesion
3.5. CCL2 and CCL5 Secreted by Macrophages Are Associated with Enhanced Ovarian Cancer-Mesothelial Cell Adhesion
4. Discussion
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
- Siegel, R.L.; Miller, K.D.; Fuchs, H.E.; Jemal, A. Cancer statistics, 2022. CA Cancer J. Clin. 2022, 72, 7–33. [Google Scholar] [CrossRef] [PubMed]
- Yung, S.; Li, F.K.; Chan, T.M. Peritoneal mesothelial cell culture and biology. Perit. Dial. Int. 2006, 26, 162–173. [Google Scholar] [CrossRef] [PubMed]
- Lengyel, E. Ovarian cancer development and metastasis. Am. J. Pathol. 2010, 177, 1053–1064. [Google Scholar] [CrossRef] [PubMed]
- Castells, M.; Thibault, B.; Delord, J.P.; Couderc, B. Implication of tumor microenvironment in chemoresistance: Tumor-associated stromal cells protect tumor cells from cell death. Int. J. Mol. Sci. 2012, 13, 9545–9571. [Google Scholar] [CrossRef] [PubMed]
- Hanahan, D.; Coussens, L.M. Accessories to the crime: Functions of cells recruited to the tumor microenvironment. Cancer Cell 2012, 21, 309–322. [Google Scholar] [CrossRef] [Green Version]
- Baghban, R.; Roshangar, L.; Jahanban-Esfahlan, R.; Seidi, K.; Ebrahimi-Kalan, A.; Jaymand, M.; Kolahian, S.; Javaheri, T.; Zare, P. Tumor microenvironment complexity and therapeutic implications at a glance. Cell Commun. Signal. 2020, 18, 59. [Google Scholar] [CrossRef] [Green Version]
- Ribeiro Franco, P.I.; Rodrigues, A.P.; de Menezes, L.B.; Pacheco Miguel, M. Tumor microenvironment components: Allies of cancer progression. Pathol. Res. Pract. 2020, 216, 152729. [Google Scholar] [CrossRef]
- Zhang, M.; He, Y.; Sun, X.; Li, Q.; Wang, W.; Zhao, A.; Di, W. A high M1/M2 ratio of tumor-associated macrophages is associated with extended survival in ovarian cancer patients. J. Ovarian Res. 2014, 7, 19. [Google Scholar] [CrossRef] [Green Version]
- Colvin, E.K. Tumor-associated macrophages contribute to tumor progression in ovarian cancer. Front. Oncol. 2014, 4, 137. [Google Scholar] [CrossRef] [Green Version]
- Yuan, X.; Zhang, J.; Li, D.; Mao, Y.; Mo, F.; Du, W.; Ma, X. Prognostic significance of tumor-associated macrophages in ovarian cancer: A meta-analysis. Gynecol. Oncol. 2017, 147, 181–187. [Google Scholar] [CrossRef]
- Etzerodt, A.; Moulin, M.; Doktor, T.K.; Delfini, M.; Mossadegh-Keller, N.; Bajenoff, M.; Sieweke, M.H.; Moestrup, S.K.; Auphan-Anezin, N.; Lawrence, T. Tissue-resident macrophages in omentum promote metastatic spread of ovarian cancer. J. Exp. Med. 2020, 217, e20191869. [Google Scholar] [CrossRef] [PubMed]
- Hensler, M.; Kasikova, L.; Fiser, K.; Rakova, J.; Skapa, P.; Laco, J.; Lanickova, T.; Pecen, L.; Truxova, I.; Vosahlikova, S.; et al. M2-like macrophages dictate clinically relevant immunosuppression in metastatic ovarian cancer. J. Immunother. Cancer 2020, 8, e000979. [Google Scholar] [CrossRef] [PubMed]
- Jeong, M.; Wang, Y.Y.; Choi, J.Y.; Lim, M.C.; Choi, J.H. CC Chemokine Ligand 7 Derived from Cancer-Stimulated Macrophages Promotes Ovarian Cancer Cell Invasion. Cancers 2021, 13, 2745. [Google Scholar] [CrossRef]
- Langmead, B.; Salzberg, S.L. Fast gapped-read alignment with Bowtie 2. Nat. Methods 2012, 9, 357–359. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Quinlan, A.R.; Hall, I.M. BEDTools: A flexible suite of utilities for comparing genomic features. Bioinformatics 2010, 26, 841–842. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Sherman, B.T.; Hao, M.; Qiu, J.; Jiao, X.; Baseler, M.W.; Lane, H.C.; Imamichi, T.; Chang, W. DAVID: A web server for functional enrichment analysis and functional annotation of gene lists (2021 update). Nucleic Acids Res. 2022, 50, W216–W221. [Google Scholar] [CrossRef]
- Chen, J.; Liu, N.N.; Li, J.Q.; Yang, L.; Zeng, Y.; Zhao, X.M.; Xu, L.L.; Luo, X.; Wang, B.; Wang, X.R. Association between ITGA2 C807T polymorphism and gastric cancer risk. World J. Gastroenterol. 2011, 17, 2860–2866. [Google Scholar] [CrossRef]
- Hisamatsu, T.; Mabuchi, S.; Sasano, T.; Kuroda, H.; Takahashi, R.; Matsumoto, Y.; Kawano, M.; Kozasa, K.; Takahashi, K.; Sawada, K.; et al. The significance of lymphatic space invasion and its association with vascular endothelial growth factor-C expression in ovarian cancer. Clin. Exp. Metastasis 2015, 32, 789–798. [Google Scholar] [CrossRef]
- Karolchik, D.; Hinrichs, A.S.; Kent, W.J. The UCSC Genome Browser. Curr. Protoc. Bioinform. 2012, 40, 1–4. [Google Scholar] [CrossRef]
- Fujita, H.; Fujita, T.; Fujii, H. IL-3-Induced Immediate Expression of c-fos and c-jun Is Modulated by the IKK2-JNK Axis. Cells 2022, 11, 1451. [Google Scholar] [CrossRef]
- Jin, J.K.; Lin, J.T.; Xu, A.K.; Lou, J.A.; Qian, C.; Li, X.M.; Wang, Y.T.; Yu, W.; Tao, H.M. CCL2: An Important Mediator Between Tumor Cells and Host Cells in Tumor Microenvironment. Front. Oncol. 2021, 11, 722916. [Google Scholar] [CrossRef] [PubMed]
- Kadomoto, S.; Izumi, K.; Mizokami, A. Roles of CCL2-CCR2 Axis in the Tumor Microenvironment. Int. J. Mol. Sci. 2021, 22, 8530. [Google Scholar] [CrossRef] [PubMed]
- Aldinucci, D.; Borghese, C.; Casagrande, N. The CCL5/CCR5 Axis in Cancer Progression. Cancers 2020, 12, 1765. [Google Scholar] [CrossRef] [PubMed]
- Huang, R.; Wang, S.; Wang, N.; Zheng, Y.; Zhou, J.; Yang, B.; Wang, X.; Zhang, J.; Guo, L.; Wang, S.; et al. CCL5 derived from tumor-associated macrophages promotes prostate cancer stem cells and metastasis via activating beta-catenin/STAT3 signaling. Cell Death Dis. 2020, 11, 234. [Google Scholar] [CrossRef] [Green Version]
- Wilson, A.P. Mesothelial cells stimulate the anchorage-independent growth of human ovarian tumour cells. Br. J. Cancer 1989, 59, 876–882. [Google Scholar] [CrossRef] [Green Version]
- Mutsaers, S.E.; Prele, C.M.; Pengelly, S.; Herrick, S.E. Mesothelial cells and peritoneal homeostasis. Fertil. Steril. 2016, 106, 1018–1024. [Google Scholar] [CrossRef] [Green Version]
- Fujikake, K.; Kajiyama, H.; Yoshihara, M.; Nishino, K.; Yoshikawa, N.; Utsumi, F.; Suzuki, S.; Niimi, K.; Sakata, J.; Mitsui, H.; et al. A novel mechanism of neovascularization in peritoneal dissemination via cancer-associated mesothelial cells affected by TGF-beta derived from ovarian cancer. Oncol. Rep. 2018, 39, 193–200. [Google Scholar] [CrossRef] [Green Version]
- Kenny, H.A.; Chiang, C.Y.; White, E.A.; Schryver, E.M.; Habis, M.; Romero, I.L.; Ladanyi, A.; Penicka, C.V.; George, J.; Matlin, K.; et al. Mesothelial cells promote early ovarian cancer metastasis through fibronectin secretion. J. Clin. Investig. 2014, 124, 4614–4628. [Google Scholar] [CrossRef] [Green Version]
- Valls, A.F.; Shen, Y.; Schmidt, T. A core of macrophages facilitates ovarian cancer metastases. Transl. Cancer Res. 2017, 6, S189–S196. [Google Scholar] [CrossRef]
- Arango Duque, G.; Descoteaux, A. Macrophage cytokines: Involvement in immunity and infectious diseases. Front. Immunol. 2014, 5, 491. [Google Scholar] [CrossRef]
- Steitz, A.M.; Steffes, A.; Finkernagel, F.; Unger, A.; Sommerfeld, L.; Jansen, J.M.; Wagner, U.; Graumann, J.; Muller, R.; Reinartz, S. Tumor-associated macrophages promote ovarian cancer cell migration by secreting transforming growth factor beta induced (TGFBI) and tenascin C. Cell Death Dis. 2020, 11, 249. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- O’Connor, T.; Heikenwalder, M. CCL2 in the Tumor Microenvironment. Adv. Exp. Med. Biol. 2021, 1302, 1–14. [Google Scholar] [CrossRef] [PubMed]
- Li, D.; Ji, H.; Niu, X.; Yin, L.; Wang, Y.; Gu, Y.; Wang, J.; Zhou, X.; Zhang, H.; Zhang, Q. Tumor-associated macrophages secrete CC-chemokine ligand 2 and induce tamoxifen resistance by activating PI3K/Akt/mTOR in breast cancer. Cancer Sci. 2020, 111, 47–58. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Yang, Y.I.; Wang, Y.Y.; Ahn, J.H.; Kim, B.H.; Choi, J.H. CCL2 overexpression is associated with paclitaxel resistance in ovarian cancer cells via autocrine signaling and macrophage recruitment. Biomed. Pharmacother. 2022, 153, 113474. [Google Scholar] [CrossRef] [PubMed]
- Yasui, H.; Kajiyama, H.; Tamauchi, S.; Suzuki, S.; Peng, Y.; Yoshikawa, N.; Sugiyama, M.; Nakamura, K.; Kikkawa, F. CCL2 secreted from cancer-associated mesothelial cells promotes peritoneal metastasis of ovarian cancer cells through the P38-MAPK pathway. Clin. Exp. Metastasis 2020, 37, 145–158. [Google Scholar] [CrossRef]
- Carroll, M.J.; Fogg, K.C.; Patel, H.A.; Krause, H.B.; Mancha, A.S.; Patankar, M.S.; Weisman, P.S.; Barroilhet, L.; Kreeger, P.K. Alternatively-Activated Macrophages Upregulate Mesothelial Expression of P-Selectin to Enhance Adhesion of Ovarian Cancer Cells. Cancer Res. 2018, 78, 3560–3573. [Google Scholar] [CrossRef] [Green Version]
- Anderson, L.R.; Owens, T.W.; Naylor, M.J. Integrins in development and cancer. Biophys. Rev. 2014, 6, 191–202. [Google Scholar] [CrossRef] [Green Version]
- Dhaliwal, D.; Shepherd, T.G. Molecular and cellular mechanisms controlling integrin-mediated cell adhesion and tumor progression in ovarian cancer metastasis: A review. Clin. Exp. Metastasis 2022, 39, 291–301. [Google Scholar] [CrossRef]
- Shen, L.I.; Liu, L.; Yang, Z.; Jiang, N. Identification of genes and signaling pathways associated with squamous cell carcinoma by bioinformatics analysis. Oncol. Lett. 2016, 11, 1382–1390. [Google Scholar] [CrossRef] [Green Version]
- Huang, Y.L.; Liang, C.Y.; Ritz, D.; Coelho, R.; Septiadi, D.; Estermann, M.; Cumin, C.; Rimmer, N.; Schotzau, A.; Nunez Lopez, M.; et al. Collagen-rich omentum is a premetastatic niche for integrin alpha2-mediated peritoneal metastasis. Elife 2020, 9, e59442. [Google Scholar] [CrossRef]
- Gavalas, N.G.; Liontos, M.; Trachana, S.P.; Bagratuni, T.; Arapinis, C.; Liacos, C.; Dimopoulos, M.A.; Bamias, A. Angiogenesis-related pathways in the pathogenesis of ovarian cancer. Int. J. Mol. Sci. 2013, 14, 15885–15909. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Yang, H.; Kim, C.; Kim, M.J.; Schwendener, R.A.; Alitalo, K.; Heston, W.; Kim, I.; Kim, W.J.; Koh, G.Y. Soluble vascular endothelial growth factor receptor-3 suppresses lymphangiogenesis and lymphatic metastasis in bladder cancer. Mol. Cancer 2011, 10, 36. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Kong, D.; Zhou, H.; Neelakantan, D.; Hughes, C.J.; Hsu, J.Y.; Srinivasan, R.R.; Lewis, M.T.; Ford, H.L. VEGF-C mediates tumor growth and metastasis through promoting EMT-epithelial breast cancer cell crosstalk. Oncogene 2021, 40, 964–979. [Google Scholar] [CrossRef] [PubMed]
- Kim, I.; Moon, S.O.; Kim, S.H.; Kim, H.J.; Koh, Y.S.; Koh, G.Y. Vascular endothelial growth factor expression of intercellular adhesion molecule 1 (ICAM-1), vascular cell adhesion molecule 1 (VCAM-1), and E-selectin through nuclear factor-kappa B activation in endothelial cells. J. Biol. Chem. 2001, 276, 7614–7620. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Vlahakis, N.E.; Young, B.A.; Atakilit, A.; Sheppard, D. The lymphangiogenic vascular endothelial growth factors VEGF-C and -D are ligands for the integrin alpha9beta1. J. Biol. Chem. 2005, 280, 4544–4552. [Google Scholar] [CrossRef] [Green Version]
- Xu, S.; Zhang, T.; Cao, Z.; Zhong, W.; Zhang, C.; Li, H.; Song, J. Integrin-alpha9beta1 as a Novel Therapeutic Target for Refractory Diseases: Recent Progress and Insights. Front. Immunol. 2021, 12, 638400. [Google Scholar] [CrossRef]
- Wei, P.L.; Huang, C.Y.; Tai, C.J.; Batzorig, U.; Cheng, W.L.; Hunag, M.T.; Chang, Y.J. Glucose-regulated protein 94 mediates metastasis by CCT8 and the JNK pathway in hepatocellular carcinoma. Tumour Biol. 2016, 37, 8219–8227. [Google Scholar] [CrossRef]
- Chang, M.C.; Chen, C.A.; Chen, P.J.; Chiang, Y.C.; Chen, Y.L.; Mao, T.L.; Lin, H.W.; Lin Chiang, W.H.; Cheng, W.F. Mesothelin enhances invasion of ovarian cancer by inducing MMP-7 through MAPK/ERK and JNK pathways. Biochem. J. 2012, 442, 293–302. [Google Scholar] [CrossRef] [Green Version]
- Eriksson, M.; Arminen, L.; Karjalainen-Lindsberg, M.L.; Leppa, S. AP-1 regulates alpha2beta1 integrin expression by ERK-dependent signals during megakaryocytic differentiation of K562 cells. Exp Cell Res. 2005, 304, 175–186. [Google Scholar] [CrossRef]
- Lee, C.C.; Chen, S.C.; Tsai, S.C.; Wang, B.W.; Liu, Y.C.; Lee, H.M.; Shyu, K.G. Hyperbaric oxygen induces VEGF expression through ERK, JNK and c-Jun/AP-1 activation in human umbilical vein endothelial cells. J. Biomed. Sci. 2006, 13, 143–156. [Google Scholar] [CrossRef]
- Li, D.; Ni, S.; Miao, K.S.; Zhuang, C. PI3K/Akt and caspase pathways mediate oxidative stress-induced chondrocyte apoptosis. Cell Stress Chaperones 2019, 24, 195–202. [Google Scholar] [CrossRef] [PubMed]
- Zhu, Y.; Pan, Y.; Zhang, G.; Wu, Y.; Zhong, W.; Chu, C.; Qian, Y.; Zhu, G. Chelerythrine Inhibits Human Hepatocellular Carcinoma Metastasis in Vitro. Biol. Pharm. Bull. 2018, 41, 36–46. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Yang, J.; Nie, J.; Ma, X.; Wei, Y.; Peng, Y.; Wei, X. Targeting PI3K in cancer: Mechanisms and advances in clinical trials. Mol. Cancer 2019, 18, 26. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Gasparri, M.L.; Bardhi, E.; Ruscito, I.; Papadia, A.; Farooqi, A.A.; Marchetti, C.; Bogani, G.; Ceccacci, I.; Mueller, M.D.; Benedetti Panici, P. PI3K/AKT/mTOR Pathway in Ovarian Cancer Treatment: Are We on the Right Track? Geburtshilfe Frauenheilkd 2017, 77, 1095–1103. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Hughes, C.E.; Nibbs, R.J.B. A guide to chemokines and their receptors. FEBS J. 2018, 285, 2944–2971. [Google Scholar] [CrossRef]
- Tyner, J.W.; Uchida, O.; Kajiwara, N.; Kim, E.Y.; Patel, A.C.; O’Sullivan, M.P.; Walter, M.J.; Schwendener, R.A.; Cook, D.N.; Danoff, T.M.; et al. CCL5-CCR5 interaction provides antiapoptotic signals for macrophage survival during viral infection. Nat. Med. 2005, 11, 1180–1187. [Google Scholar] [CrossRef]
- Fei, L.; Ren, X.; Yu, H.; Zhan, Y. Targeting the CCL2/CCR2 Axis in Cancer Immunotherapy: One Stone, Three Birds? Front. Immunol. 2021, 12, 771210. [Google Scholar] [CrossRef]
- Osborn, G.; Stavraka, C.; Adams, R.; Sayasneh, A.; Ghosh, S.; Montes, A.; Lacy, K.E.; Kristeleit, R.; Spicer, J.; Josephs, D.H.; et al. Macrophages in ovarian cancer and their interactions with monoclonal antibody therapies. Clin. Exp. Immunol. 2022, 209, 4–21. [Google Scholar] [CrossRef]
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Cho, S.-K.; Lee, K.; Woo, J.-H.; Choi, J.-H. Macrophages Promote Ovarian Cancer-Mesothelial Cell Adhesion by Upregulation of ITGA2 and VEGFC in Mesothelial Cells. Cells 2023, 12, 384. https://doi.org/10.3390/cells12030384
Cho S-K, Lee K, Woo J-H, Choi J-H. Macrophages Promote Ovarian Cancer-Mesothelial Cell Adhesion by Upregulation of ITGA2 and VEGFC in Mesothelial Cells. Cells. 2023; 12(3):384. https://doi.org/10.3390/cells12030384
Chicago/Turabian StyleCho, Seung-Kye, Kijun Lee, Jeong-Hwa Woo, and Jung-Hye Choi. 2023. "Macrophages Promote Ovarian Cancer-Mesothelial Cell Adhesion by Upregulation of ITGA2 and VEGFC in Mesothelial Cells" Cells 12, no. 3: 384. https://doi.org/10.3390/cells12030384
APA StyleCho, S. -K., Lee, K., Woo, J. -H., & Choi, J. -H. (2023). Macrophages Promote Ovarian Cancer-Mesothelial Cell Adhesion by Upregulation of ITGA2 and VEGFC in Mesothelial Cells. Cells, 12(3), 384. https://doi.org/10.3390/cells12030384