Cullin 3 overexpression inhibits lung cancer metastasis and is associated with survival of lung adenocarcinoma
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.
KeywordsCullin 3 Lung cancer Nrf2 RhoA Cell proliferation Metastasis Prognosis
Non-small cell lung cancer
Cullin-RING ubiquitin ligases
Nuclear factor erythroid 2-related factor
We thank all members of our study team and the included participants for their cooperation.
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.
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.
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.
The authors declare that they agree to submit the article for publication.
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