Silencing of cyclooxygenase‑2 inhibits the growth, invasion and migration of ovarian cancer cells

Lin,Yang; Cui,Manhua; Xu,Tianmin; Yu,Wei; Zhang,Lihui

doi:10.3892/mmr.2014.2131

Silencing of cyclooxygenase‑2 inhibits the growth, invasion and migration of ovarian cancer cells

Authors:
- Yang Lin
- Manhua Cui
- Tianmin Xu
- Wei Yu
- Lihui Zhang
View Affiliations

Affiliations: Department of Gynaecology and Obstetrics, The Second Hospital of Jilin University, Changchun, Jilin 130041, P.R. China
Published online on: April 9, 2014 https://doi.org/10.3892/mmr.2014.2131
Pages: 2499-2504

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

The present study aimed to investigate the effect of downregulating cyclooxygenase‑2 (COX‑2) expression on the growth of human ovarian cancer cells. The COX‑2‑specific small interfering RNA (siRNA) plasmid vector was constructed and then transfected into ovarian cancer cells. The expression of COX‑2 mRNA and protein was detected by quantitative polymerase chain reaction and western blot analysis, respectively. Cell proliferation, apoptosis, cell cycle distribution and cell migration were assessed following knockdown of COX‑2 by RNA interference (RNAi). Western blot analysis was used to identify differentially expressed angiogenesis- and cell cycle‑associated proteins in cells with silenced COX‑2. The expression levels of COX‑2 in ovarian cancer cells transfected with siRNA were decreased, leading to a significant inhibition of ovarian cancer cell proliferation, migration and invasion. Western blot analysis revealed that silencing of COX‑2 may inhibit vascular endothelial growth factor, matrix metalloproteinase (MMP)‑2 and MMP‑9 protein expression. In conclusion, the present study demonstrated that RNAi can effectively silence COX‑2 gene expression and inhibit the growth of ovarian cancer cells, which indicates that there is a potential of targeting COX‑2 as a novel gene therapy approach for the treatment of ovarian cancer.

View Figures

View References

1	Lin Y, Peng S, Yu H, Teng H and Cui M: RNAi-mediated downregulation of NOB1 suppresses the growth and colony-formation ability of human ovarian cancer cells. Med Oncol. 29:311–317. 2012. View Article : Google Scholar : PubMed/NCBI
2	Auersperg N, Wong AS, Choi KC, Kang SK and Leung PC: Ovarian surface epithelium: biology, endocrinology, and pathology. Endocr Rev. 22:255–258. 2001.PubMed/NCBI
3	Ziebarth AJ, Landen CN Jr and Alvarez RD: Molecular/genetic therapies in ovarian cancer: future opportunities and challenges. Clin Obstet Gynecol. 55:156–172. 2012. View Article : Google Scholar : PubMed/NCBI
4	Wang H, Linghu H, Wang J, Che YL, Xiang TX, Tang WX and Yao ZW: The role of Crk/Dock180/Rac1 pathway in the malignant behavior of human ovarian cancer cell SKOV3. Tumour Biol. 31:59–67. 2010. View Article : Google Scholar : PubMed/NCBI
5	Fader AN and Rose PG: Role of surgery in ovarian carcinoma. J Clin Oncol. 25:2873–2883. 2007. View Article : Google Scholar : PubMed/NCBI
6	Salzberg M, Thurlimann B, Bonnefois H, Fink D, Rochlitz C, Von Moos R and Senn H: Current concepts of treatment strategies in advanced or recurrent ovarian cancer. Oncology. 68:293–298. 2005. View Article : Google Scholar : PubMed/NCBI
7	McGuire WP and Ozols RF: Chemotherapy of advanced ovarian cancer. Semin Oncol. 25:340–348. 1998.PubMed/NCBI
8	Fung-Kee-Fung M, Oliver T, Elit L, Oza A, Hirte HW and Bryson P: Optimal chemotherapy treatment for women with recurrent ovarian cancer. Curr Oncol. 14:195–208. 2007. View Article : Google Scholar : PubMed/NCBI
9	Dempke W, Rie C, Grothey A and Schmoll HJ: Cyclooxygenase-2: a novel target for cancer chemotherapy? J Cancer Res Clin Oncol. 127:411–417. 2001. View Article : Google Scholar : PubMed/NCBI
10	Williams CS, Mann M and DuBois RN: The role of cyclooxygenases in inflammation, cancer, and development. Oncogene. 18:7908–7916. 1999. View Article : Google Scholar : PubMed/NCBI
11	Li S, Miner K, Fannin R, Carl Barrett J and Davis BJ: Cyclooxygenase-1 and 2 in normal and malignant human ovarian epithelium. Gynecol Oncol. 92:622–627. 2004. View Article : Google Scholar
12	Denkert C, Köbel M, Pest S, Koch I, Berger S, Schwabe M, Siegert A, Reles A, Klosterhalfen B and Hauptmann S: Expression of cyclooxygenase-2 is an independent prognostic factor in human ovarian carcinoma. Am J Pathol. 160:893–903. 2002. View Article : Google Scholar : PubMed/NCBI
13	Erkinheimo TL, Lassus H, Finne P, van Rees BP, Leminen A, Ylikorkala O, Haglund C, Butzow R and Ristimäki A: Elevated cyclooxygenase-2 expression is associated with altered expression of p53 and SMAD4, amplification of HER-2/neu, and poor outcome in serous ovarian carcinoma. Clin Cancer Res. 10:538–545. 2004. View Article : Google Scholar : PubMed/NCBI
14	Arico S, Pattingre S, Bauvy C, Gane P, Barbat A, Codogno P and Ogier-Denis E: Celecoxib induces apoptosis by inhibiting 3-phosphoinositide-dependent protein kinase-1 activity in the human colon cancer HT-29 cell line. J Biol Chem. 277:27613–27621. 2002. View Article : Google Scholar : PubMed/NCBI
15	Cerutti H: RNA interference: traveling in the cell and gaining functions? Trends Genet. 19:39–46. 2003. View Article : Google Scholar : PubMed/NCBI
16	Fire A, Xu S, Montgomery MK, Kostas SA, Driver SE and Mello CC: Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature. 391:806–811. 1998. View Article : Google Scholar : PubMed/NCBI
17	Hannon GL and Rossi JJ: Unlocking the potential of the human genome with RNA interference. Nature. 431:371–378. 2004. View Article : Google Scholar : PubMed/NCBI
18	Izquierdo M: Short interfering RNAs as a tool for cancer gene therapy. Cancer Gene Ther. 12:217–227. 2005. View Article : Google Scholar : PubMed/NCBI
19	Elbashir SM, Harborth J, Lendeckel W, Yalcin A, Weber K and Tuschl T: Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells. Nature. 411:494–498. 2001. View Article : Google Scholar : PubMed/NCBI
20	Caplen NJ: Gene therapy progress and prospects. Downregulating gene expression: the impact of RNA interference. Gene Ther. 11:1241–1248. 2004. View Article : Google Scholar : PubMed/NCBI
21	Elbashir SM, Harborth J, Weber K and Tuschl T: Analysis of gene function in somatic mammalian cells using small interfering RNAs. Methods. 26:199–213. 2002. View Article : Google Scholar : PubMed/NCBI
22	Tai MH, Weng CH, Mon DP, Hu CY and Wu MH: Ultraviolet C irradiation induces different expression of cyclooxygenase 2 in NIH 3T3 cells and A431 cells: the roles of COX-2 are different in various cell lines. Int J Mol Sci. 13:4351–4366. 2012. View Article : Google Scholar : PubMed/NCBI
23	Kim DH, Longo M, Han Y, Lundberg P, Cantin E and Rossi JJ: Interferon induction by siRNAs and ssRNAs synthesized by phage polymerase. Nat Biotechnol. 22:321–325. 2004. View Article : Google Scholar : PubMed/NCBI
24	Meister G and Tuschi T: Mechanisms of gene silencing by double-stranded RNA. Nature. 431:343–349. 2004. View Article : Google Scholar : PubMed/NCBI
25	Mello CC and Conte D Jr: Revealing the world of RNA interference. Nature. 431:338–342. 2004. View Article : Google Scholar : PubMed/NCBI
26	Wang R, Wang X, Lin F, Gao P, Dong K and Zhang HZ: shRNA-targeted cyclooxygenase (COX)-2 inhibits proliferation, reduces invasion and enhances chemosensitivity in laryngeal carcinoma cells. Mol Cell Biochem. 317:179–188. 2008. View Article : Google Scholar
27	Chen Q, Pan Q, Cai R and Qian C: Prospects of RNA interference induced by RNA Pol II promoter in cancer therapy. Prog Biochem Biophys. 34:806–815. 2007.
28	Strillacci A, Griffoni C, Spisni E, Manara MC and Tomasi V: RNA interference as a key to knockdown overexpressed cyclooxygenase-2 gene in tumour cells. Br J Cancer. 94:1300–1310. 2006. View Article : Google Scholar : PubMed/NCBI
29	Vogler M, Walczak H, Stadel D, Haas TL, Genze F, Jovanovic M, Gschwend JE, Simmet T, Debatin KM and Fulda S: Targeting XIAP bypasses Bcl-2-mediated resistance to TRAIL and cooperates with TRAIL to suppress pancreatic cancer growth in vitro and in vivo. Cancer Res. 68:7956–7965. 2008. View Article : Google Scholar : PubMed/NCBI
30	Zhang L, Wu YD, Li P, Tu J, Niu YL, Xu CM and Zhang ST: Effects of cyclooxygenase-2 on human esophageal squamous cell carcinoma. World J Gastroenterol. 17:4572–4580. 2011. View Article : Google Scholar : PubMed/NCBI
31	Denkert C, Fürstenberg A, Daniel PT, Koch I, Köbel M, Weichert W, Siegert A and Hauptmann S: Induction of G0/G1 cell cycle arrest in ovarian carcinoma cells by the anti-inflammatory drug NS-398, but not by COX-2-specific RNA interference. Oncogene. 22:8653–8661. 2003. View Article : Google Scholar : PubMed/NCBI
32	Park W, Oh YT, Han JH and Pyo H: Antitumor enhancement of celecoxib, a selective cyclooxygenase-2 inhibitor, in a Lewis lung carcinoma expressing cyclooxygenase-2. J Exp Clin Cancer Res. 27:662008. View Article : Google Scholar : PubMed/NCBI
33	Müller-Decker K and Fürstenberger G: The cyclooxygenase-2-mediated prostaglandin signaling is causally related to epithelial carcinogenesis. Mol Carcinog. 46:705–710. 2007.PubMed/NCBI
34	Leahy KM, Ornberg RL, Wang Y, Zweifel BS, Koki AT and Masferrer JL: Cyclooxygenase-2 inhibition by celecoxib reduces proliferation and induces apoptosis in angiogenic endothelial cells in vivo. Cancer Res. 62:625–631. 2002.PubMed/NCBI
35	Mamdani M, Juurlink DN, Lee DS, Rochon PA, Koop A, Naglie G, Austin PC, Laupacis A and Stukel TA: Cyclo-oxygenase-2 inhibitors versus non-selective non-steroidal anti-inflammatory drugs and congestive heart failure outcomes in elderly patients: a population-based cohort study. Lancet. 363:1751–1756. 2004. View Article : Google Scholar : PubMed/NCBI
36	Rasmuson A, Kock A, Fuskevåg OM, Kruspig B, Simón-Santamaría J, Gogvadze V, Johnsen JI, Kogner P and Sveinbjörnsson B: Autocrine prostaglandin E2 signaling promotes tumor cell survival and proliferation in childhood neuroblastoma. PLoS One. 7:e293312012. View Article : Google Scholar : PubMed/NCBI
37	Pockaj BA, Basu GD, Pathangey LB, Gray RJ, Hernandez JL, Gendler SJ and Mukherjee P: Reduced T-cell and dendritic cell function is related to cyclooxygenase-2 overexpression and prostaglandin E2 secretion in patients with breast cancer. Ann Surg Oncol. 11:328–339. 2004. View Article : Google Scholar : PubMed/NCBI
38	Iñiguez MA, Rodríguez A, Volpert OV, Fresno M and Redondo JM: Cyclooxygenase-2: a therapeutic target in angiogenesis. Trends Mol Med. 9:73–78. 2003.