Binding between ROCK1 and DCTN2 triggers diabetes‑associated centrosome amplification in colon cancer cells

Li,Yuan Fei; Shi,Lin Jie; Wang,Pu; Wang,Jia Wen; Shi,Guang Yi; Lee,Shao Chin

doi:10.3892/or.2021.8102

Binding between ROCK1 and DCTN2 triggers diabetes‑associated centrosome amplification in colon cancer cells

Authors:
- Yuan Fei Li
- Lin Jie Shi
- Pu Wang
- Jia Wen Wang
- Guang Yi Shi
- Shao Chin Lee
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Affiliations: Department of Oncology, The First Hospital, Shanxi Medical University, Taiyuan, Shanxi 030001, P.R. China, Changzhi Medical University, Changzhi, Shanxi 030001, P.R. China, Institute of Biomedical Sciences of The School of Life Sciences, Jiangsu Normal University, Xuzhou, Jiangsu 221116, P.R. China
Published online on: June 2, 2021 https://doi.org/10.3892/or.2021.8102
Article Number: 151
Copyright: © Li et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Type 2 diabetes increases the risk various types of cancer and is associated with a poor prognosis therein. There is also evidence that the disease is associated with cancer metastasis. Centrosome amplification can initiate tumorigenesis with metastasis in vivo and increase the invasiveness of cancer cells in vitro. Our previous study reported that type 2 diabetes promotes centrosome amplification via the upregulation and centrosomal translocation of Rho‑associated protein kinase 1 (ROCK1), which suggests that centrosome amplification is a candidate biological link between type 2 diabetes and cancer development. In the present study, functional proteomics analysis was used to further investigate the molecular pathways underlying centrosome amplification by targeting ROCK1 binding partners. High glucose, insulin and palmitic acid were used to induce centrosome amplification, and immunofluorescent staining was employed to visualize centrosomal alterations. Combined with immunoprecipitation, mass spectrometry‑based proteomics analysis was used to identify ROCK1 binding proteins, and protein complex disruption was achieved by siRNA‑knockdown. In total, 1,148 ROCK1 binding proteins were identified, among which 106 proteins were exclusively associated with the treated samples, 193 were only associated with the control samples, and 849 were found in both the control and treated samples. Of the proteins with evidence of centrosomal localization, Dynactin subunit 2 (DCTN2) was confirmed to be localized to the centrosomes. Treating the cells with high glucose, insulin and palmitic acid increased the protein levels of ROCK1 and DCTN2, promoted their binding with each other, and triggered centrosome amplification. Disruption of the protein complex by knocking down ROCK1 or DCTN2 expression partially attenuated centrosome amplification, while simultaneous knockdown of both proteins completely inhibited centrosome amplification. These results suggested ROCK1‑DCTN2 binding as a signal for the regulation of centrosome homeostasis, which is key for diabetes‑associated centrosome amplification, and enriches our knowledge of centrosome biology. Therefore, the ROCK1‑DCTN2 complex may serve as a target for inhibiting centrosome amplification both in research or future therapeutic development.

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