Abstract | BACKGROUND: METHODS: We used CRISPR/Cas9 gene editing to knock-out plectin in B16 mouse melanoma cells. Protein levels of plectin and Src activity were examined by western blotting analysis. In vivo tumor formation was assessed by subcutaneous injection of B16 cells into nude mice and histological analysis performed after 2 weeks by Hematoxylin- Eosin (H&E) staining. Cell proliferation was evaluated by direct cell count, cell counting kit-8 assays, cyclin D1 mRNA expression and Ki-67 immunostaining. Cell aggregation and adhesion were examined by spheroid formation, dispase-based dissociation assay and cell adhesion assays. RESULTS: In in vivo tumor formation assays, depletion of plectin resulted in low-density tumors with large intercellular spaces. In vitro experiments revealed that plectin-deficient B16 cells exhibit reduced cell proliferation and reduced cell-to-cell adhesion. Since Src activity is reduced in plectin-deficient melanomas, we examined the relationship between plectin and Src signaling. Src overexpression in plectin knockout B16 cells rescued cell proliferation and improved cell-to-cell adhesion and cell to extracellular matrix adhesion. CONCLUSION: These results suggest that plectin plays critical roles in tumor formation by promoting cell proliferation and cell-to-cell adhesion through Src signaling activity in melanoma cells.
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Authors | Kana Mizuta, Takuma Matsubara, Akino Goto, William N Addison, Mitsushiro Nakatomi, Kou Matsuo, Yukiyo Tada-Shigeyama, Tatsuki Yaginuma, Hiromi Honda, Izumi Yoshioka, Shoichiro Kokabu |
Journal | BMC cancer
(BMC Cancer)
Vol. 22
Issue 1
Pg. 936
(Aug 30 2022)
ISSN: 1471-2407 [Electronic] England |
PMID | 36038818
(Publication Type: Journal Article)
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Copyright | © 2022. The Author(s). |
Chemical References |
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Topics |
- Animals
- Cell Line, Tumor
- Cell Movement
- Cell Proliferation
- Melanoma, Experimental
(metabolism)
- Mice
- Mice, Nude
- Oncogenes
- Plectin
(genetics)
- Sarcoma, Avian
(genetics)
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