Browse > Article
http://dx.doi.org/10.7314/APJCP.2015.16.4.1343

Knockdown of UHRF1 by Lentivirus-mediated shRNA Inhibits Ovarian Cancer Cell Growth  

Yan, Feng (College of Chemistry and Chemical Engineering, Yangzhou University)
Shao, Li-Jia (Department of clinical Laboratory, Nanjing Medical University Cancer Hospital & Jiangsu Cancer Hospital)
Hu, Xiao-Ya (College of Chemistry and Chemical Engineering, Yangzhou University)
Publication Information
Asian Pacific Journal of Cancer Prevention / v.16, no.4, 2015 , pp. 1343-1348 More about this Journal
Abstract
Human UHRF1 (ubiquitin-like PHD and RING finger domain-containing 1) has been reported to be over-expressed in many cancers, but its role in ovarian cancer remains elusive. Here, we determined whether knockdown of UHRF1 by lentivirus-mediated shRNA could inhibit ovarian cancer cell growth. Lentivirus-mediated short hairpin RNAs (lv-shRNAs-UHRF1) were designed to trigger the gene silencing RNA interference (RNAi) pathway. The efficiency of lentivirus-mediated shRNA infection into HO-8910 and HO-8910 PM cells was determined using fluorescence microscopy to observe lentivirus-mediated GFP expression and was confirmed to be over 80 percent. UHRF1 expression in infected HO-8910 and HO-8910 PM was evaluated by real-time PCR and Western blot analysis. The Cell Counting Kit-8 (CCK-8) assay was used to measure cell viability; flow cytometry and Hoechst 33342 assay was applied to measure cell cycle arrest and apoptosis. Cell invasion was assessed using transwell chambers. Our results demonstrated that the loss of UHRF1 promoted HO-8910 and HO-8910 PM cell apoptosis, while inhibiting cell proliferation. In addition, UHRF1 knockdown significantly inhibited the invasion of human ovarian cancer cells. In the present study, we also showed that depleting HO-8910 cells of UHRF1 caused activation of the DNA damage response pathway, with the cell cycle arrested in G2/M-phase. The DNA damage response in cells depleted of UHRF1 was illustrated by phosphorylation of CHK (checkpoint kinase) 2 on Thr68, phosphorylation of CDC25 (cell division control 25) on Ser 216 and phosphorylation of CDK1 (cyclin-dependent kinase 1) on Tyr 15.
Keywords
Ovarian cancer cells; UHRF1; apoptosis; cell cycle; phosphorylation; transcription factors;
Citations & Related Records
연도 인용수 순위
  • Reference
1 Abbady AQ, Bronner C, Bathami K, et al (2005). TCR pathway involves ICBP90 gene down-regulation via E2F binding sites. Biochem Pharmacol, 70, 570-9.   DOI
2 Achour M, Mousli M, Alhosin M, et al (2013). Epigallocatechin-3-gallate up-regulates tumor suppressor gene expression via a reactive oxygen species-dependent down-regulation of UHRF1. Biochem Biophys Res Commun, 430, 208-12.   DOI
3 Alhosin M, Sharif T, Mousli M, et al (2011). Down-regulation of UHRF1, associated with re-expression of tumor suppressor genes, is a common feature of natural compounds exhibiting anti-cancer properties. J Exp Clin Cancer Res, 30, 41.   DOI
4 Arima Y, Hirota T, Bronner C, et al (2004). Down-regulation of nuclear protein ICBP90 by p53/p21Cip1/WAF1-dependent DNA-damage checkpoint signals contributes to cell cycle arrest at G1/S transition. Genes to Cells, 9, 131-42.   DOI
5 Dandache I, et al (2012). Anti-cancer properties of cranberry juice in human colon cancer: Characterization of the cellular and molecular mechanisms. Fundament Clin Pharmacol, 26, 40.
6 Fang L, Shanqu L, Ping G, et al (2012). Gene therapy with RNAi targeting UHRF1 driven by tumor-specific promoter inhibits tumor growth and enhances the sensitivity of chemotherapeutic drug in breast cancer in vitro and in vivo. Cancer Chemotherapy Pharmacol, 69, 1079-87.   DOI
7 Geng Y, Gao Y, Ju H, et al (2012). Diagnostic and prognostic value of plasma and tissue ubiquitin-like, containing PHD and RING finger domains 1 in breast cancer patients. Cancer Science, 104, 194-9.
8 Jenkins Y, Markovtsov V, Lang W, et al (2005). Critical role of the ubiquitin ligase activity of UHRF1, a nuclear RING finger protein, in tumor cell growth. Molecular Biology of the Cell, 16, 5621-9.   DOI
9 Mousli M, Hopfner R, Abbady AQ, et al (2013). ICBP90 belongs to a new family of proteins with an expression that is deregulated in cancer cells. Br J Cancer, 89, 120-7.
10 Tien AL, Senbanerjee S, Kulkarni A, et al (2011). UHRF1 depletion causes a G2/M arrest, activation of DNA damage response and apoptosis. Biochemical J, 435, 175-85.   DOI
11 Wang F, Yang YZ, Shi CZ, et al (2012). UHRF1 promotes cell growth and metastasis through repression of p16 ink4a in colorectal cancer. Ann Surgical Oncol, 19, 2753-62.   DOI
12 Yang G.L, Zhang LH, Bo JJ, et al (2012). UHRF1 is associated with tumor recurrence in non-muscle-invasive bladder cancer. Medical Oncol, 29, 842-7.   DOI