Effect of UBE2Q on BECN and CEA Protein in Colorectal Cancer Cells

Abstract

Background: Expression of UBE2Q1, UBE2Q2, and members of the ubiquitinconjugating enzyme family (E2) are affected in colorectal cancer (CRC). The BECN gene plays a key role in CRC cells. In gastrointestinal carcinoma therapy, tumorassociated antigens such as CEA are typically used.To investigate the association
between UBE2Q1 and Beclin1 autophagy marker and CEA protein expression in LS180 CRC cell line.
Materials and Methods: In this study, changes in the expression of BECN marker in LS180 cell lines with the vector containing UBE2Q1 were investigated using real-time PCR. The expression of CEA protein was also evaluated by western blotting. Statistical analyses were performed with Graph Pad Prism software.
Results: The results indicated reduced expression of BECN autophagy marker (P=0). Therefore, in the presence of UBE2Q1, cancer cells have less ability to induce autophagy. However, CEA protein levels in LS180 transfected cells with a UBE2Q1- ORF-containing plasmid decreased when compared to non-transfected cells.
Conclusion: The use of pharmacologic inhibitors related to the autophagy mechanism can be a novel approach in cancer therapy.

References
[1] Friedemann-Sánchez G, Griffin JM, Partin MR. Gender differences in colorectal cancer screening barriers and information needs 1. Health Expect. 2007;10(2):148-60.

[2] Smith RA, Cokkinides V, Eyre HJ. American Cancer Society guidelines for the early detection of cancer, 2003. CA Cancer J Clin. 2003;53(1):27-43.

[3] De Pergola G, Silvestris F. Obesity as a major risk factor for cancer. J Obes. 2013;2013.

[4] Fuchs CS, Giovannucci EL, Colditz GA, Hunter DJ, Speizer FE, Willett WC. A prospective study of family history and the risk of colorectal cancer. N Engl J Med. 1994;331(25):1669-74.

[5] Garraway LA, Lander ES. Lessons from the cancer genome. Cell. 2013; 153(1):17-37.

[6] Loos B, Engelbrecht AM, Lockshin RA, Klionsky DJ, Zakeri Z. The variability of autophagy and cell death susceptibility: Unanswered questions. Autophagy. 2013:9(9):1270-85.

[7] Jiang X, Overholtzer M, Thompson CB. Autophagy in cellular metabolism and cancer. J Clin Invest. 2015; 125:47-54.

[8] Kenific CM, Debnath J. Cellular and metabolic functions for autophagy in cancer cells. Trends Cell Biol. 2015; 25(1): 37-45.

[9] Shafiee SM, Seghatoleslam A, Nikseresht M, Hosseini SV, Alizadeh-Naeeni M, Safaei A, Owji AA. UBE2Q1 expression in human colorectal tumors and cell lines. Mol Biol Rep. 2013; 40(12): 7045-51.

[10] Chang R, Wei L, Lu Y, Cui X, Lu C, Liu L. Upregulated expression of ubiquitin-conjugating enzyme E2Q1 (UBE2Q1) is associated with enhanced cell proliferation and poor prognosis in human hapatocellular
carcinoma. J Mol Histol. 2015; 46(1): 45-56.

[11] Brooks CL, Gu W. Ubiquitination, phosphorylation and acetylation: the molecular basis for p53 regulation. Curr Opin Cell Biol. 2003; 15(2): 164-71.

[12] Kang R, Zeh HJ, Lotze MT, Tang D. The Beclin 1 network regulates autophagy and apoptosis. Cell Death Differ. 2011; 18(4): 571-80.

[13] Yu L, McPhee CK, Zheng L, Mardones GA, Rong Y, Peng J. Termination of autophagy and reformation of lysosomes regulated by mTOR. Nature. 2010; 465(7300):942-6.

[14] Pihl E, Nairn RC, Hughes ES, Cuthbertson AM, Rollo AJ. Mucinous colorectal carcinoma: immunopathology and prognosis. Pathology. 1980; 12(3):439-47.

[15] Wagener C, Müller-Wallraf R, Nisson S, Gröner J, Breuer H. Localization and concentration of carcinoembryonic antigen (CEA) in gastrointestinal tumors: correlation with CEA levels in plasma. J Natl Cancer Inst. 1981; 67(3): 539-47.

[16] Nikseresht M, Seghatoleslam A, Monabati A, Talei A, Ghalati FB, Owji AA. Overexpression of the novel human gene, UBE2Q2, in breast cancer. Cancer Genet Cytogenet. 2010;197(2):101-6.

[17] Mrayam R. Study the effect of UBE2Q1over expression on the protein level of p53 and the expression of cyclin E,BCL2 and MDM2 genes in LS180 and SW480 colorectal cancer cell lines: Shiraz University of Medical Sience; 2015.

[18] Garraway LA, Lander ES. Lessons from the cancer genome. Cell. 2013; 153(1):17-37.

[19] Galluzzi L, Pietrocola F, Bravo-San Pedro JM, Amaravadi RK, Baehrecke EH, Cecconi F, et al. Autophagy in malignant transformation and cancer progression. EMBO J. 2015; 34(7):856-80.

[20] Chang R, Wei L, Lu Y, Cui X, Lu C, Liu L, et al. Upregulated expression of ubiquitin-conjugating enzyme E2Q1 (UBE2Q1) is associated with enhanced cell proliferation and poor prognosis in human hapatocellular carcinoma. J Mol Histol. 2015; 46(1):45-56.

[21] Schmitz KJ, Ademi C, Bertram S, Schmid KW, Baba HA. Prognostic relevance of autophagy-related markers LC3, p62/sequestosome 1, Beclin-1 and ULK1 in colorectal cancer patients with respect to KRAS mutational status. World J Surg Oncol. 2016; 14(1):189.

[22] Wu S, Sun C, Tian D, Li Y, Gao X, He S, Li T. Expression and clinical significances of Beclin1, LC3 and mTOR in colorectal cancer. Int J Clin Exp Pathol. 2015; 8(4): 3882.

[23] Nicholson BD, Shinkins B, Pathiraja I, Roberts NW, James TJ, Mallett S, Perera R, Primrose JN, Mant
D. Blood CEA levels for detecting recurrent colorectal cancer. Cochrane Database Syst Rev. 2015(12): CD011134.

[24] Palos G, Gilmore KR, Chapman P, Lewis-Patterson P, Bi W, Rodriguez MA. Low concordance with CEA
tumor marker monitoring in colorectal cancer survivors. Cancer Res. 2016; 7(14): 3464.