A Novel Approach to Reducing Lung Metastasis in Osteosarcoma: Increasing Cell Stiffness with Carbenoxolone
Aim: Primary malignant bone tumor osteosarcoma can metastasize to the lung. Diminishing lung metastasis would positively affect the prognosis of patients. Our previous studies demonstrated that highly metastatic osteosarcoma cell lines are significantly softer than low-metastasis cell lines. We ther...
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MDPI AG
2023-05-01
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| Series: | Current Issues in Molecular Biology |
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| Online Access: | https://www.mdpi.com/1467-3045/45/5/278 |
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| author | Kouji Kita Kunihiro Asanuma Takayuki Okamoto Eiji Kawamoto Koichi Nakamura Tomohito Hagi Tomoki Nakamura Motomu Shimaoka Akihiro Sudo |
| author_facet | Kouji Kita Kunihiro Asanuma Takayuki Okamoto Eiji Kawamoto Koichi Nakamura Tomohito Hagi Tomoki Nakamura Motomu Shimaoka Akihiro Sudo |
| author_sort | Kouji Kita |
| collection | DOAJ |
| description | Aim: Primary malignant bone tumor osteosarcoma can metastasize to the lung. Diminishing lung metastasis would positively affect the prognosis of patients. Our previous studies demonstrated that highly metastatic osteosarcoma cell lines are significantly softer than low-metastasis cell lines. We therefore hypothesized that increasing cell stiffness would suppress metastasis by reducing cell motility. In this study, we tested whether carbenoxolone (CBX) increases the stiffness of LM8 osteosarcoma cells and prevents lung metastasis in vivo. Methods: We evaluated the actin cytoskeletal structure and polymerization of CBX-treated LM8 cells using actin staining. Cell stiffness was measured using atomic force microscopy. Metastasis-related cell functions were analyzed using cell proliferation, wound healing, invasion, and cell adhesion assays. Furthermore, lung metastasis was examined in LM8-bearing mice administered with CBX. Results: Treatment with CBX significantly increased actin staining intensity and stiffness of LM8 cells compared with vehicle-treated LM8 cells (<i>p</i> < 0.01). In Young’s modulus images, compared with the control group, rigid fibrillate structures were observed in the CBX treatment group. CBX suppressed cell migration, invasion, and adhesion but not cell proliferation. The number of LM8 lung metastases were significantly reduced in the CBX administration group compared with the control group (<i>p</i> < 0.01). Conclusion: In this study, we demonstrated that CBX increases tumor cell stiffness and significantly reduces lung metastasis. Our study is the first to provide evidence that reducing cell motility by increasing cell stiffness might be effective as a novel anti-metastasis approach in vivo. |
| first_indexed | 2024-03-11T03:50:57Z |
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| language | English |
| last_indexed | 2024-03-11T03:50:57Z |
| publishDate | 2023-05-01 |
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| record_format | Article |
| series | Current Issues in Molecular Biology |
| spelling | doaj.art-99563cbe1ed441c886d5023c824ac1982023-11-18T00:57:17ZengMDPI AGCurrent Issues in Molecular Biology1467-30371467-30452023-05-014554375438810.3390/cimb45050278A Novel Approach to Reducing Lung Metastasis in Osteosarcoma: Increasing Cell Stiffness with CarbenoxoloneKouji Kita0Kunihiro Asanuma1Takayuki Okamoto2Eiji Kawamoto3Koichi Nakamura4Tomohito Hagi5Tomoki Nakamura6Motomu Shimaoka7Akihiro Sudo8Department of Orthopedic Surgery, Mie University Graduate School of Medicine, 2-174 Edobashi, Tsu 514-8507, Mie, JapanDepartment of Orthopedic Surgery, Mie University Graduate School of Medicine, 2-174 Edobashi, Tsu 514-8507, Mie, JapanDepartment of Pharmacology, Faculty of Medicine, Shimane University, 89-1 Enya-cho, Izumo-shi 693-8501, Shimane, JapanDepartment of Molecular Pathobiology and Cell Adhesion Biology, Mie University Graduate School of Medicine, 2-174 Edobashi, Tsu 514-8507, Mie, JapanDepartment of Orthopedic Surgery, Mie University Graduate School of Medicine, 2-174 Edobashi, Tsu 514-8507, Mie, JapanDepartment of Orthopedic Surgery, Mie University Graduate School of Medicine, 2-174 Edobashi, Tsu 514-8507, Mie, JapanDepartment of Orthopedic Surgery, Mie University Graduate School of Medicine, 2-174 Edobashi, Tsu 514-8507, Mie, JapanDepartment of Molecular Pathobiology and Cell Adhesion Biology, Mie University Graduate School of Medicine, 2-174 Edobashi, Tsu 514-8507, Mie, JapanDepartment of Orthopedic Surgery, Mie University Graduate School of Medicine, 2-174 Edobashi, Tsu 514-8507, Mie, JapanAim: Primary malignant bone tumor osteosarcoma can metastasize to the lung. Diminishing lung metastasis would positively affect the prognosis of patients. Our previous studies demonstrated that highly metastatic osteosarcoma cell lines are significantly softer than low-metastasis cell lines. We therefore hypothesized that increasing cell stiffness would suppress metastasis by reducing cell motility. In this study, we tested whether carbenoxolone (CBX) increases the stiffness of LM8 osteosarcoma cells and prevents lung metastasis in vivo. Methods: We evaluated the actin cytoskeletal structure and polymerization of CBX-treated LM8 cells using actin staining. Cell stiffness was measured using atomic force microscopy. Metastasis-related cell functions were analyzed using cell proliferation, wound healing, invasion, and cell adhesion assays. Furthermore, lung metastasis was examined in LM8-bearing mice administered with CBX. Results: Treatment with CBX significantly increased actin staining intensity and stiffness of LM8 cells compared with vehicle-treated LM8 cells (<i>p</i> < 0.01). In Young’s modulus images, compared with the control group, rigid fibrillate structures were observed in the CBX treatment group. CBX suppressed cell migration, invasion, and adhesion but not cell proliferation. The number of LM8 lung metastases were significantly reduced in the CBX administration group compared with the control group (<i>p</i> < 0.01). Conclusion: In this study, we demonstrated that CBX increases tumor cell stiffness and significantly reduces lung metastasis. Our study is the first to provide evidence that reducing cell motility by increasing cell stiffness might be effective as a novel anti-metastasis approach in vivo.https://www.mdpi.com/1467-3045/45/5/278metastasiscell stiffnessatomic force microscopycarbenoxoloneactin cytoskeleton |
| spellingShingle | Kouji Kita Kunihiro Asanuma Takayuki Okamoto Eiji Kawamoto Koichi Nakamura Tomohito Hagi Tomoki Nakamura Motomu Shimaoka Akihiro Sudo A Novel Approach to Reducing Lung Metastasis in Osteosarcoma: Increasing Cell Stiffness with Carbenoxolone Current Issues in Molecular Biology metastasis cell stiffness atomic force microscopy carbenoxolone actin cytoskeleton |
| title | A Novel Approach to Reducing Lung Metastasis in Osteosarcoma: Increasing Cell Stiffness with Carbenoxolone |
| title_full | A Novel Approach to Reducing Lung Metastasis in Osteosarcoma: Increasing Cell Stiffness with Carbenoxolone |
| title_fullStr | A Novel Approach to Reducing Lung Metastasis in Osteosarcoma: Increasing Cell Stiffness with Carbenoxolone |
| title_full_unstemmed | A Novel Approach to Reducing Lung Metastasis in Osteosarcoma: Increasing Cell Stiffness with Carbenoxolone |
| title_short | A Novel Approach to Reducing Lung Metastasis in Osteosarcoma: Increasing Cell Stiffness with Carbenoxolone |
| title_sort | novel approach to reducing lung metastasis in osteosarcoma increasing cell stiffness with carbenoxolone |
| topic | metastasis cell stiffness atomic force microscopy carbenoxolone actin cytoskeleton |
| url | https://www.mdpi.com/1467-3045/45/5/278 |
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