Induction of cell proliferation, clonogenicity and cell accumulation in S phase as a consequence of human UBE2Q1 overexpression

Fahmidehkar,Mohammad  Ali; Shafiee,Sayed  Mohammad; Eftekhar,Ebrahim; Mahbudi,Laleh; Seghatoleslam,Atefeh

doi:10.3892/ol.2016.4860

Induction of cell proliferation, clonogenicity and cell accumulation in S phase as a consequence of human UBE2Q1 overexpression

Authors:
- Mohammad Ali Fahmidehkar
- Sayed Mohammad Shafiee
- Ebrahim Eftekhar
- Laleh Mahbudi
- Atefeh Seghatoleslam
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Affiliations: Department of Biochemistry, School of Medicine, Shiraz University of Medical Sciences, Shiraz 71348-45794, Iran, Food and Cosmetic Health Research Center, Hormozgan University of Medical Sciences, Bandar Abbas 79158-73665, Iran
Published online on: July 13, 2016 https://doi.org/10.3892/ol.2016.4860
Pages: 2169-2174

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Abstract

Ubiquitination is an important cellular mechanism with a pivotal role in the degradation of abnormal or short-lived proteins and the regulation of cell cycle and cell growth. The ubiquitin-proteasome pathway is altered in multiple types of human malignancies, including colorectal cancer (CRC). The alteration in the expression of the novel human gene ubiquitin-conjugating enzyme E2 Q1 (UBE2Q1), as a putative member of the E2 ubiquitin‑conjugating enzyme family, has been reported in several malignancies, including carcinoma of the breast, hepatocellular and colorectal cancer, and pediatric acute lymphoblastic leukemia. In the present study, the effect of UBE2Q1 overexpression on cell growth, clonogenicity, motility and cell cycle was investigated in a CRC cell line. The UBE2Q1 gene was cloned in the pCMV6-AN-GFP expression vector. A series of stable transfectants of SW1116 cells overexpressing UBE2Q1 protein were established and confirmed by fluorescence microscopy and western blotting. Using these cells, MTT assay was performed to evaluate cell growth and proliferation, while crystal violet staining was used for clonogenicity assay. Cell cycle analysis was also performed to survey the ratio of cells accumulated in different phases of the cell cycle upon transfection. The motility of these cells was also studied using wound healing assay. UBE2Q1 transfectants exhibited a faster growth in cell culture, increased colony formation capacity and enhanced motility compared with control non-transfected cells and cells transfected with empty vector (mock-transfected cells). UBE2Q1 overexpression also resulted in a significant decrease in the number of cells accumulated in the G0/G1 phase of the cell cycle. The present findings suggest that UBE2Q1 may function as an oncogene that induces proliferation of cancer cells, and could be a novel diagnostic tool and a potential therapeutic target for CRC.

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1	Shen Z, Jiang X, Zeng C, Zheng S, Luo B, Zeng Y, Ding R, Jiang H, He Q, Guo J and Jie W: High expression of ubiquitin-conjugating enzyme 2C (UBE2C) correlates with nasopharyngeal carcinoma progression. BMC Cancer. 13:1922013. View Article : Google Scholar : PubMed/NCBI
2	Grzmil P, Altmann ME, Adham IM, Engel U, Jarry H, Schweyer S, Wolf S, Mänz J and Engel W: Embryo implantation failure and other reproductive defects in Ube2q1-deficient female mice. Reproduction. 145:45–56. 2013. View Article : Google Scholar : PubMed/NCBI
3	Nikseresht M, Seghatoleslam A, Monabati A, Talei A, Ghalati FB and Owji AA: Overexpression of the novel human gene, UBE2Q2, in breast cancer. Cancer Genet Cytogenet. 197:101–106. 2010. View Article : Google Scholar : PubMed/NCBI
4	Hershko A: The ubiquitin system for protein degradation and some of its roles in the control of the cell division cycle. Cell Death Differ. 12:1191–1197. 2005. View Article : Google Scholar : PubMed/NCBI
5	Zhu D, Zhong Y, Wu H, Ye L, Wang J, Li Y, Wei Y, Ren L, Xu B, Xu J and Qin X: Predicting metachronous liver metastasis from colorectal cancer using serum proteomic fingerprinting. J Surg Res. 184:861–866. 2013. View Article : Google Scholar : PubMed/NCBI
6	Zhou LH, Hu Q, Sui H, Ci SJ, Wang Y, Liu X, Liu NN, Yin PH, Qin JM and Li Q: Tanshinone II-a inhibits angiogenesis through down regulation of COX-2 in human colorectal cancer. Asian Pac J Cancer Prev. 13:4453–4458. 2012. View Article : Google Scholar : PubMed/NCBI
7	Van Cutsem E, Borràs JM, Castells A, Ciardiello F, Ducreux M, Haq A, Schmoll HJ and Tabernero J: Improving outcomes in colorectal cancer: Where do we go from here? Eur J Cancer. 49:2476–2485. 2013. View Article : Google Scholar : PubMed/NCBI
8	Kopetz S, Chang GJ, Overman MJ, Eng C, Sargent DJ, Larson DW, Grothey A, Vauthey JN, Nagorney DM and McWilliams RR: Improved survival in metastatic colorectal cancer is associated with adoption of hepatic resection and improved chemotherapy. J Clin Oncol. 27:3677–3683. 2009. View Article : Google Scholar : PubMed/NCBI
9	Chang R, Wei L, Lu Y, Cui X, Lu C, Liu L, Jiang D, Xiong Y, Wang G, Wan C, 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. 46:45–56. 2015. View Article : Google Scholar : PubMed/NCBI
10	Chen Z, He X, Jia M, Liu Y, Qu D, Wu D, Wu P, Ni C, Zhang Z, Ye J, et al: β-catenin overexpression in the nucleus predicts progress disease and unfavourable survival in colorectal cancer: A meta-analysis. PLoS One. 8:e638542013. View Article : Google Scholar : PubMed/NCBI
11	Bai R, Li D, Shi Z, Fang X, Ge W and Zheng S: Clinical significance of Ankyrin repeat domain 12 expression in colorectal cancer. J Exp Clin Cancer Res. 32:352013. View Article : Google Scholar : PubMed/NCBI
12	Seghatoleslam A, Nikseresht M, Shafiee SM, Monabati A, Namavari MM, Talei A, Safaei A and Owji AA: Expression of the novel human gene, UBE2Q1, in breast tumors. Mol Biol Rep. 39:5135–5141. 2012. View Article : Google Scholar : PubMed/NCBI
13	Shafiee SM, Seghatoleslam A, Nikseresht M, Hosseini SV, Alizadeh-Naeeni M, Safaei A and Owji AA: UBE2Q1 expression in human colorectal tumors and cell lines. Mol Biol Rep. 40:7045–7051. 2013. View Article : Google Scholar : PubMed/NCBI
14	Seghatoleslam A, Zambrano A, Millon R, Ganguli G, Argentini M, Cromer A, Abecassis J and Wasylyk B: Analysis of a novel human gene, LOC92912, overexpressed in hypopharyngeal tumours. Biochem Biophys Res Commun. 339:422–429. 2006. View Article : Google Scholar : PubMed/NCBI
15	Seghatoleslam A, Monabati A, Bozorg-Ghalati F, Nikseresht M, Bordbar MR, Rahvar M and Owji AA: Expression of UBE2Q2, a putative member of the ubiquitin-conjugating enzyme family in pediatric acute lymphoblastic leukemia. Arch Iran Med. 15:352–355. 2012.PubMed/NCBI
16	Shafiee SM, Seghatoleslam A, Nikseresht M, Hosseini SV, Alizadeh-Naeeni M, Safaei A and Owji AA: Expression status of UBE2Q2 in colorectal primary tumors and cell lines. Iran J Med Sci. 39(2 Suppl): S196–S202. 2014.
17	Zhu S, Yao F, Li WH, Wan JN, Zhang YM, Tang Z, Khan S, Wang CH and Sun SR: PKC?-dependent activation of the ubiquitin proteasome system is responsible for high glucose-induced human breast cancer MCF-7 cell proliferation, migration and invasion. Asian Pac J Cancer Prev. 14:5687–5692. 2013. View Article : Google Scholar : PubMed/NCBI
18	Burger AM and Seth AK: The ubiquitin-mediated protein degradation pathway in cancer: Therapeutic implications. Eur J Cancer. 40:2217–2229. 2004. View Article : Google Scholar : PubMed/NCBI
19	Voutsadakis IA: Ubiquitin- and ubiquitin-like proteins-conjugating enzymes (E2s) in breast cancer. Mol Biol Rep. 40:2019–2034. 2013. View Article : Google Scholar : PubMed/NCBI
20	Hao Z, Zhang H and Cowell J: Ubiquitin-conjugating enzyme UBE2C: Molecular biology, role in tumorigenesis, and potential as a biomarker. Tumour Biol. 33:723–730. 2012. View Article : Google Scholar : PubMed/NCBI
21	Seghatoleslam A and Zambrano A: Effects of overexpression of LOC92912 gene on cell cycle progression. Iran J Med Sci. 34:277–284. 2009.
22	Tang XK, Wang KJ, Tang YK and Chen L: Effects of ubiquitin-conjugating enzyme 2C on invasion, proliferation and cell cycling of lung cancer cells. Asian Pac J Cancer Prev. 15:3005–3009. 2014. View Article : Google Scholar : PubMed/NCBI
23	Chen S, Chen Y, Hu C, Jing H, Cao Y and Liu X: Association of clinicopathological features with UbcH10 expression in colorectal cancer. J Cancer Res Clin Oncol. 136:419–426. 2010. View Article : Google Scholar : PubMed/NCBI
24	Pallante P, Berlingieri MT, Troncone G, Kruhoffer M, Orntoft TF, Viglietto G, Caleo A, Migliaccio I, Decaussin-Petrucci M, Santoro M, et al: UbcH10 overexpression may represent a marker of anaplastic thyroid carcinomas. Br J Cancer. 93:464–471. 2005. View Article : Google Scholar : PubMed/NCBI
25	Ieta K, Ojima E, Tanaka F, Nakamura Y, Haraguchi N, Mimori K, Inoue H, Kuwano H and Mori M: Identification of overexpressed genes in hepatocellular carcinoma, with special reference to ubiquitin-conjugating enzyme E2C gene expression. Int J Cancer. 121:33–38. 2007. View Article : Google Scholar : PubMed/NCBI
26	Berlingieri MT, Pallante P, Sboner A, Barbareschi M, Bianco M, Ferraro A, Mansueto G, Borbone E, Guerriero E, Troncone G and Fusco A: UbcH10 is overexpressed in malignant breast carcinomas. Eur J Cancer. 43:2729–2735. 2007. View Article : Google Scholar : PubMed/NCBI
27	Bose MV, Gopisetty G, Selvaluxmy G and Rajkumar T: Dominant negative Ubiquitin-conjugating enzyme E2C sensitizes cervical cancer cells to radiation. Int J Radiat Biol. 88:629–634. 2012. View Article : Google Scholar : PubMed/NCBI
28	Perrotta I, Bruno L, Maltese L, Russo E, Donato A and Donato G: Immunohistochemical analysis of the ubiquitin-conjugating enzyme UbcH10 in lung cancer: A useful tool for diagnosis and therapy. J Histochem Cytochem. 60:359–365. 2012. View Article : Google Scholar : PubMed/NCBI
29	Lin J, Raoof DA, Wang Z, Lin MY, Thomas DG, Greenson JK, Giordano TJ, Orringer MB, Chang AC, Beer DG and Lin L: Expression and effect of inhibition of the ubiquitin-conjugating enzyme E2C on esophageal adenocarcinoma. Neoplasia. 8:1062–1071. 2006. View Article : Google Scholar : PubMed/NCBI
30	van Wijk SJ and Timmers HT: The family of ubiquitin-conjugating enzymes (E2s): Deciding between life and death of proteins. FASEB J. 24:981–993. 2010. View Article : Google Scholar : PubMed/NCBI
31	Seghatoleslam A, Bozorg-Ghalati F, Monabati A, Nikseresht M and Owji AA: UBE2Q1, as a down regulated gene in pediatric acute lymphoblastic leukemia. Int J Mol Cell Med. 3:95–101. 2014.PubMed/NCBI
32	Wan C, Chen J, Hu B, Zou H, Li A, Guo A and Jiang J: Downregulation of UBE2Q1 is associated with neuronal apoptosis in rat brain cortex following traumatic brain injury. J Neurosci Res. 92:1–12. 2014. View Article : Google Scholar : PubMed/NCBI
33	Shafiee SM, Rasti M, Seghatoleslam A, Azimi T and Owji AA: UBE2Q1 in a Human Breast Carcinoma Cell Line: Overexpression and Interaction with p53. Asian Pac J Cancer Prev. 16:3723–7. 2015. View Article : Google Scholar : PubMed/NCBI
34	Grelle G, Kostka S, Otto A, Kersten B, Genser KF, Müller EC, Wälter S, Böddrich A, Stelzl U, Hänig C, et al: Identification of VCP/p97, carboxyl terminus of Hsp70-interacting protein (CHIP), and amphiphysin II interaction partners using membrane-based human proteome arrays. Mol Cell Proteomics. 5:234–244. 2006. View Article : Google Scholar : PubMed/NCBI
35	Min JN, Whaley RA, Sharpless NE, Lockyer P, Portbury AL and Patterson C: CHIP deficiency decreases longevity, with accelerated aging phenotypes accompanied by altered protein quality control. Mol Cell Biol. 28:4018–4025. 2008. View Article : Google Scholar : PubMed/NCBI
36	Naito AT, Okada S, Minamino T, Iwanaga K, Liu ML, Sumida T, Nomura S, Sahara N, Mizoroki T, Takashima A, et al: Promotion of CHIP-mediated p53 degradation protects the heart from ischemic injury. Circ Res. 106:1692–1702. 2010. View Article : Google Scholar : PubMed/NCBI
37	Banerjee S, Brooks WS and Crawford DF: Inactivation of the ubiquitin conjugating enzyme UBE2Q2 causes a prophase arrest and enhanced apoptosis in response to microtubule inhibiting agents. Oncogene. 26:6509–6517. 2007. View Article : Google Scholar : PubMed/NCBI
38	Melner MH, Haas AL, Klein JM, Brash AR, Boeglin WE, Nagdas SK, Winfrey VP and Olson GE: Demonstration of ubiquitin thiolester formation of UBE2Q2 (UBCi), a novel ubiquitin-conjugating enzyme with implantation site-specific expression. Biol Reprod. 75:395–406. 2006. View Article : Google Scholar : PubMed/NCBI