Expression of Deubiquitinating Enzymes in Lung Cancer Cells with Different REST Status

  • Wedad Alkut Biomedical science, University of Liverpool, Crown Street, Liverpool, L69 3BX, UK. Department of Biology, Faculty of Sciences, El-Khoms, El-mergib University.
Keywords: NSCLC, SCLC, Deubiquitinating DUBs, REST.

Abstract

The aims of this study were: (1) to screen a panel of lung cell lines for expression of the proteins UCHL1, USP49, USP15, USP4, and USP11. (2)  To investigate the correlation of transcript and protein expression level of these DUBs. (3) To find any proteins that discriminate non-small cell lung cancer (NSCLC) from neuroendocrine (NE) SCLC or benign lung carcinoid. (4) To investigate any correlation of DUB proteins expression with lung cancer sub-types that has different amounts of REST protein.

Method: Western blots were performed to detect the expression of DUB expression (USP4, USP11, USP15, USP49, and UCHL1) and REST protein in extracts from normal lung cell lines, SCLC, carcinoid, and NSCLC

Results: we found that the transcript level was in general not a good indicator of DUB protein level. There was a significant positive correlation of REST with DUB proteins from whole-cell extracts for USP15, USP11, USP4, and USP49 in normal lung cell lines.  The P values were 0.016, 0.002, 0 .018 and 0 .034 respectively. Also in both normal and NSCLC cell lines total USP15, USP11 and USP49 showed a positive correlation with REST. P values were 0.007, 0.029 and 0.001 respectively. However USP11 protein was highly abundant in REST-deficient SCLC. In contrast, both USP11 and USP15 showed a positive correlation with REST in SCLC and NSCLC nuclear extracts. P values were 0.001 and 0.005 respectively. A positive correlation of DUBs with REST would be consistent with their putative role in stabilizing REST.

References

Amerik, A. Y., & Hochstrasser, M. (2004). Mechanism and function of deubiquitinating enzymes. Biochim Biophys Acta, 1695(1-3), 189-207.

Bührens, R. A., Amelung, J. T., Reymond, M. A., & Beshay, M. (2009). Protein Expression in Human Non-Small Cell Lung Cancer: A Systematic Database. Pathobiology, 76, 277-285.

Celis, J. E., Kruhoffer, M., Gromova, I., Frederiksen, C., Ostergaard, M., Thykjaer, T., . . . Orntoft, T. F. (2000). Gene expression profiling: monitoring transcription and translation products using DNA microarrays and proteomics. FEBS Lett, 480, 2–16.

Coulson, J. M. (2005). Transcriptional Regulation: Cancer, Neurons and the REST. Curr Bio, 15, 65-66.

Coulson, J. M., & et al. (2003). Detection of small cell lung cancer by RT-PCR for neuropeptides, neuropeptide receptors, or a splice variant of the neuron restrictive silencer factor. Methods Mol Med,, 75, 335-352.

Coulson, J. M., Ocejo-Garcia, & Woll, P. J. (2003). Neuroendocrine phenotype of small cell lung cancer. Methods Mol Med,, 74, 61-73.

Daviet, L., & et al. (2008). Targeting ubiqutin specific proteases for drug discovery. Biochimie, 90, 270-283.

Gray, D. A., Inazawa, J., Gupta, K., Wong, A., Ueda, R., & Takahashi, T. (1995). Elevated expression of Unph, a proto-oncogene at 3p21.3, in human lung tumors. Oncogene. 10, :2179–2183.

Guardavaccaro, D., et al,. (2008). Control of chromosome stability by the β-TrCP–REST–Mad2 axis. Nature, 452:, 365–369. .

Haglund, K., & Dikic, I. (2005). Ubiquitylation and cell signaling. EMBO J, 24, 3353 – 3359.

Huang, X., et al. (2009). The COP9 signalosome mediates beta-catenin degradation by deneddylation and blocks adenomatous polyposis coli destruction via USP15. J Mol Biol,, 391(4), 691-702.

Komander, D., Clague, M. J., & Urbe, S. (2009). Breaking the chains: Structure and function of the deubiquitinases. Nat Rev Mol Cell Biol, 10, 550-563.

Lin, C. H., H.S. Chang, and W.C. Yu. (2008). USP11 stabilizes HPV-16E7 and further modulates the E7 biological activity. J Biol Chem, 283(23), 15681-15688.

Reports-Mortality., C. S. (February 2007). Cancer Stats Reports-Mortality. Retrieved from UK:

Sacco, J. J., et al., . (2010). Emerging roles of deubiquitinases in cancer-associated pathways. IUBMB Life, 62(2), 140-157.

Shebl, F. M., Pinto, L. A., Garcia-Pineres, A., Lempicki, R., Williams, M., Harro, C., & Hildesheim, A. (2010). Cancer Epidemiol Biomarkers Prev.

Vos, R. M., et al, . (2009). The ubiquitin-specific peptidase USP15 regulates human papillomavirus type 16 E6 protein stability. J Virol, 83(17), 8885-8892.

Westbrook, T. F., & et al. (2008). SCF-TRCP controls oncogenic transformation and neural differentiation through REST degradation. Nature, 452, 370-470.

Westbrook, T. F., Martin, E. S., & Schlabach, M. R. e. a. (2005). A genetic screen for candidate tumor suppressors identifies REST. Cell, 121, 837-848.

Yamaguchi, T., et al,. (2007). The deubiquitinating enzyme USP11 controls an IkappaB kinase alpha (IKKalpha)-p53 signaling pathway in response to tumor necrosis factor alpha (TNFalpha). J Biol Chem, 282(47), 33943-33948.

Published
2022-04-17
How to Cite
[1]
Alkut, W. 2022. Expression of Deubiquitinating Enzymes in Lung Cancer Cells with Different REST Status. Scientific Journal for the Faculty of Science-Sirte University. 2, 1 (Apr. 2022), 63-70. DOI:https://doi.org/10.37375/sjfssu.v2i1.217.
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