Cdc42 Regulates the Expression of Cytoskeleton and Microtubule Network Proteins to Promote Invasion and Metastasis of Progeny Cells Derived from CoCl2-induced Polyploid Giant Cancer Cells

Minying Zheng; Lankai Chen; Junjie Fu; Xiaohui Yang; Shuo Chen; Wenzheng Fu; Yuwei Li; Shiwu Zhang

doi:10.7150/jca.85032

Cdc42 Regulates the Expression of Cytoskeleton and Microtubule Network Proteins to Promote Invasion and Metastasis of Progeny Cells Derived from CoCl₂-induced Polyploid Giant Cancer Cells

J Cancer. 2023 Jun 26;14(10):1920-1934. doi: 10.7150/jca.85032. eCollection 2023.

Authors

Minying Zheng^{1

2}, Lankai Chen³, Junjie Fu¹, Xiaohui Yang³, Shuo Chen⁴, Wenzheng Fu⁴, Yuwei Li⁴, Shiwu Zhang¹

Affiliations

¹ Department of Pathology, Tianjin Union Medical Center, Nankai University, Tianjin, 300121, P.R. China.
² State Key Laboratory of Medicinal Chemical Biology, NanKai University, Tianjin, 300071, P.R. China.
³ Nankai University School of Medicine, Nankai University, Tianjin, 300071, P.R. China.
⁴ Department of Colorectal Surgery, Tianjin Union Medical Center, Tianjin, 300121, P.R. China.

Abstract

Purpose: Our previous studies have shown that CoCl₂ can induce the formation of polyploid giant cancer cells (PGCCs) and PGCCs could produce progeny cells via asymmetric division. In this study, the molecular mechanism by which PGCCs generate progeny cells with high invasion and migration abilities was explored. Methods: In this study, PGCCs induced by CoCl₂ produced progeny cells via asymmetric division, which was observed dynamically using laser scanning confocal microscopy. Cell cycle in LoVo and Hct116 before and after CoCl₂ treatment was analyzed by flow cytometry. Cell function experiments, co-immunoprecipitation, mass spectrometry analysis, ML141 treatment, western blotting, and siRNA transfection experiments were used to demonstrate that Cdc42/PAK1 was involved in the regulation of cytoskeleton expression. The proliferation, migration, and invasion abilities of PGCCs and progeny cells were compared in PGCCs and progeny cells with and without inhibiting the expression of Cdc42 and PAK1. Results: G2/M phase arrest appeared in CoCl_2-treated LoVo and Hct116 cells. After CoCl₂ treatment, an increased expression of Cdc42 and PAK1 led to a decrease in the expression of stathmin and an increase in the expression of phosphorylated stathmin, which is located in the nucleus of PGCCs and progeny cells. PTPN14 negatively regulates the expression of PAK1 and p38MAPK. Low levels of PTPN14 expression, a downstream regulatory protein of stathmin, endows progeny tumor cells generated by PGCCs with the ability to invade and metastasize. The expression of PKA1α, cathepsin B, and D increased in CoCl₂-treated cells compared with that in the control cells, associated with the infiltration and migration of PGCCs with their progeny cells. Conclusion: CoCl₂-induced overexpression of Cdc42 plays a critical role in increasing the infiltration and migration abilities of PGCCs and progeny cells by regulating cytoskeleton protein expression.

Keywords: Cdc42; PAK1; PGCCs; cathepsin B; cathepsin D; stathmin.