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Cullin 3 overexpression inhibits lung cancer metastasis and is associated with survival of lung adenocarcinoma

  • Jiayu Zhou
  • Shizhen Zhang
  • Yong Xu
  • Weiwen Ye
  • Zhijun Li
  • Zhoumiao Chen
  • Zhengfu HeEmail author
Research Paper

Abstract

Cullin 3 (CUL3), a molecular scaffold of Cullin-RING ubiquitin ligase, plays an important role in regulating biological processes through modulating the ubiquitylation and degradation of various protein substrates. Dysfunction of CUL3 is implicated in the development of several human diseases. However, the clinical significance and prognostic value of CUL3 in lung cancer have not been investigated. This study investigated the CUL3-modulating potential of non-small cell lung cancer cell lines, H1299, H358, H2170 and H520, by using immunoblotting, MTT, migration, invasion, colony formation and in vivo tumorigenicity assays. The prognostic significance of CUL3 was measured by public KM plotter database (http://kmplot.com/analysis/index.php?p=service&cancer=breast) and tissue immunohistochemistry analysis. The public online database analysis revealed that elevated mRNA expression of CUL3 was associated with better prognosis for non-small cell lung cancer and lung adenocarcinoma. In vitro experiments showed that ectopic overexpression of CUL3 significantly inhibited lung adenocarcinoma cell proliferation and migration, and the tumor-suppressive effect of CUL3 was dependent on the Nrf2/RhoA axis. In vivo mice model demonstrated that overexpression of CUL3 lead to a significant reduction of lung adenocarcinoma growth and metastasis. Importantly, tissue immunohistochemistry analysis showed that about 47% of non-small cell lung cancer tissues were expressed of CUL3 at high levels. Overexpression of CUL3 predicted favorable overall survival in non-small cell lung cancer patients, especially in lung adenocarcinoma, but not in lung squamous cell carcinoma patients. CUL3 could serve as a prognostic biomarker for lung adenocarcinoma. Loss of CUL3 might be driving tumorigenesis by activating the Nrf2/RhoA pathway.

Keywords

Cullin 3 Lung cancer Nrf2 RhoA Cell proliferation Metastasis Prognosis 

Abbreviations

CUL3

Cullin 3

NSCLC

Non-small cell lung cancer

OS

Overall survival

CRLs

Cullin-RING ubiquitin ligases

Nrf2

Nuclear factor erythroid 2-related factor

Notes

Acknowledgements

We thank all members of our study team and the included participants for their cooperation.

Author contributions

JZ and SZ designed and performed the experiments, analyzed and interpreted the data, and drafted the manuscript. JZ, SZ, ZC, ZH, and YX performed the experiments. WY analyzed the data. JZ, SZ, YX and ZL revised the manuscript. All authors reviewed the manuscript.

Funding

National Natural Science Foundation of China, Grant/Award Number: 81471874.

Compliance with ethical standards

Conflict of interest

All authors declare that there is no competing interest.

Ethics approval

Clinical samples were collected from Sir Run Run Shaw Hospital of Zhejiang University (Hangzhou City, China). All enrolled patients gave written informed consent for their tissues to be used for scientific research. The study was approved by the Ethics Committee of the Sir Run Run Shaw Hospital, consistent with the recommendations of the declaration of Helsinki for biomedical research, and followed standard institutional protocol for human research. Moreover, the animal study protocol was approved by the Animal Care and User Committee at Sir Run Run Shaw Hospital.

Informed consent

The authors declare that they agree to submit the article for publication.

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Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.Department of Thoracic Surgery, Sir Run Run Shaw Hospital, School of MedicineZhejiang UniversityHangzhouChina
  2. 2.Institute of Translational MedicineZhejiang University School of MedicineHangzhouChina

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