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Acetylation-stabilized chloride intracellular channel 1 exerts a tumor-promoting effect on cervical cancer cells by activating NF-κB

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Abstract

Purpose

Cervical cancer remains a major cause of cancer-related death in women, especially in developing countries. Previously, we found that the acetylation levels of chloride intracellular channel 1 (CLIC1) at lysine 131 were increased in cervical cancer tissues using a label-free proteomics approach. The aim of this study was to further determine the role of CLIC1 expression and its acetylation in cervical cancer.

Methods

CLIC1 expression and its implications for the prognosis of cervical cancer were analyzed using primary patient samples and cells, and the Gene Expression Profiling Interactive Analysis (GEPIA) database (gepia.cancer-pku.cn). The effect of CLIC1 on cervical cancer cells was evaluated using Cell Counting Kit (CCK)-8, flow cytometry, scratch wound healing, transwell, Western blotting and co-immunoprecipitation (Co-IP) assays. In vivo tumor growth was assessed using mouse xenograft models.

Results

We found that CLIC1 expression was increased in cervical cancer tissues and cells and that patients with a high CLIC1 expression tended to have a shorter overall survival time. Knockdown of CLIC1 significantly reduced in vitro cervical cancer cell proliferation, migration and invasion, and in vivo tumorigenesis. At the molecular level, we found that nuclear factor kappa B (NF-κB) activity was positively regulated by CLIC1. Pyrrolidine dithiocarbamate (PDTC), an inhibitor of NF-κB, attenuated the tumor-promoting effect of CLIC1. Moreover, we found that CLIC1 acetylation at K131 was upregulated in cervical cancer cells, which stabilized CLIC1 by inhibiting its ubiquitynation. Substitution of K131 inhibited CLIC1 ubiquitynation and promoted in vitro cervical cancer cell proliferation, migration and invasion, and in vivo tumor growth. In addition, we found that acetyltransferase HAT1 was responsible for CLIC1 acetylation at K131.

Conclusion

Our data indicate that CLIC1 acts as a tumor promoter in cervical cancer, suggesting a potential treatment strategy for cervical cancer by regulating CLIC1 expression and/or acetylation.

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Acknowledgments

This study was supported by grants from the National Natural Science Foundation of China (No. 81772274), the Natural Science Foundation of Heilongjiang Province (No. H2017045) and the Key Project of the Petrel Foundation of Harbin Medical University Cancer Hospital (No. JJZD2020-07). In addition, the study was supported by the Beijing Medical Award Foundation (No.YXJL-2020-0417-0043 and No.YXJL-2020-0510-0044) and the Heilongjiang University student innovation and Entrepreneurship training project (No.202011230002).

Availability of data and materials

The analyzed data sets generated during the study are available from the corresponding author upon reasonable request.

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Author notes

  1. Wanyue Wang, Xin Li and Ye Xu contributed equally to this work.

Authors and Affiliations

  1. School of Basic Medical Sciences, Qiqihar Medical University, Qiqihar, 161006, Heilongjiang, China

    Wanyue Wang

  2. Department of Oncology, Harbin Medical University Cancer Hospital, Harbin, 150081, Heilongjiang, China

    Xin Li

  3. Department of Gynecology, Harbin Medical University Cancer Hospital, No. 150 Haping Road, Nangang District, Harbin, 150081, Heilongjiang Province, China

    Ye Xu, Weikang Guo, Lu Zhang, Yaoxian Wang & Xiuwei Chen

  4. Department of Cardiopulmonary Function, Harbin Medical University Cancer Hospital, Harbin, 150081, Heilongjiang, China

    Hui Yu

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  1. Wanyue Wang
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Contributions

WYX and CXW conceived and designed the study. WWY, LX and XY conducted the experiments. WWY, GWK, YH and ZL analyzed the data. WWY and LX wrote the manuscript. WYX and CXW modified the manuscript. All authors read and approved the final manuscript.

Corresponding authors

Correspondence to Yaoxian Wang or Xiuwei Chen.

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Ethics approval and consent to participate

The study was approved by the Ethics Committee of the Harbin Medical University and written informed consent was obtained from all patients. All animal experiments were performed in line with the Guide for the Care and Use of Laboratory Animal by the National Institutes of Health.

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Not applicable.

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The authors declare no competing interests.

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Wang, W., Li, X., Xu, Y. et al. Acetylation-stabilized chloride intracellular channel 1 exerts a tumor-promoting effect on cervical cancer cells by activating NF-κB. Cell Oncol. 44, 557–568 (2021). https://doi.org/10.1007/s13402-020-00582-w

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