Knockdown of RhoA expression alters ovarian cancer biological behavior in vitro and in nude mice

Wang,Xiaoxia; Jiang,Wenyan; Kang,Jiali; Liu,Qicai; Nie,Miaoling

doi:10.3892/or.2015.4009

Knockdown of RhoA expression alters ovarian cancer biological behavior in vitro and in nude mice

Authors:
- Xiaoxia Wang
- Wenyan Jiang
- Jiali Kang
- Qicai Liu
- Miaoling Nie
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Affiliations: Department of Obstetrics and Gynecology, The First Affiliated Hospital of Jinan University, Guangzhou, Guangdong 510630, P.R. China, Department of Obstetrics and Gynecology, Guangzhou First People's Hospital Affiliated to Guangzhou Medical University, Guangzhou, Guangdong 510180, P.R. China, Experimental Medical Research Center, Guangzhou Medical University, Guangzhou, Guangdong 510182, P.R. China
Published online on: May 28, 2015 https://doi.org/10.3892/or.2015.4009
Pages: 891-899

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Abstract

RhoA regulates cell proliferation, migration, angiogenesis and gene expression. Altered RhoA activity contributes to cancer progression. The present study investigated the effects of RhoA knockdown on the regulation of ovarian cancer biological behavior in vitro and in nude mice. The expression of RhoA was knocked down using a lentivirus carrying RhoA short hairpin RNA (shRNA) in ovarian cancer cells and was confirmed by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and western blot analysis. The altered ovarian cancer biological behaviors were assayed by cell viability, terminal deoxynucleotidyltransferase-mediated dUTP nick end-labeling (TUNEL), migration, invasion, and nude mice tumorigenicity assays, while the altered gene expression was detected by RT-qPCR and western blot analysis. The results showed that lentivirus‑carrying RhoA shRNA significantly suppressed RhoA expression in ovarian cancer cells, which suppressed tumor cell viability, migration, invasion and adhesion in vitro. RhoA silencing also inhibited the tumorigenicity of ovarian cancer cells in nude mice, which was characterized by the suppression of tumor xenograft formation and growth and induction of tumor cell apoptosis. The results of the present study demonstrated that knockdown of RhoA expression had a significant antitumor effect on ovarian cancer cells in vitro and in nude mice, suggesting that RhoA may be a target for the development of a novel therapeutic strategy in the control of ovarian cancer.

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1	Lozano R, Naghavi M, Foreman K, Lim S, Shibuya K, Aboyans V, Abraham J, Adair T, Aggarwal R, Ahn SY, et al: Global and regional mortality from 235 causes of death for 20 age groups in 1990 and 2010: a systematic analysis for the Global Burden of Disease Study 2010. Lancet. 380:2095–2128. 2012. View Article : Google Scholar : PubMed/NCBI
2	Moyer VA: USA Preventive Services Task Force: Screening for ovarian cancer: USA Preventive Services Task Force reaffirmation recommendation statement. Ann Intern Med. 157:900–904. 2012. View Article : Google Scholar : PubMed/NCBI
3	Vo C and Carney ME: Ovarian cancer hormonal and environmental risk effect. Obstet Gynecol Clin North Am. 34:687–700. 2007. View Article : Google Scholar : PubMed/NCBI
4	Hunn J and Rodriguez GC: Ovarian cancer: etiology, risk factors, and epidemiology. Clin Obstet Gynecol. 55:3–23. 2012. View Article : Google Scholar : PubMed/NCBI
5	Gilbert L, Basso O, Sampalis J, Karp I, Martins C, Feng J, Piedimonte S, Quintal L, Ramanakumar AV and Takefman J; DOvE Study Group: Assessment of symptomatic women for early diagnosis of ovarian cancer: results from the prospective DOvE pilot project. Lancet Oncol. 13:285–291. 2012. View Article : Google Scholar : PubMed/NCBI
6	Hennessy BT, Coleman RL and Markman M: Ovarian cancer. Lancet. 374:1371–1382. 2009. View Article : Google Scholar : PubMed/NCBI
7	Hu J, Shao S, Song Y, Zhao J, Dong Y, Gong L and Yang P: Hepatocyte growth factor induces invasion and migration of ovarian cancer cells by decreasing the expression of E-cadherin, beta-catenin, and caveolin-1. Anat Rec (Hoboken). 293:1134–1139. 2010. View Article : Google Scholar
8	Fader AN and Rose PG: Role of surgery in ovarian carcinoma. J Clin Oncol. 25:2873–2883. 2007. View Article : Google Scholar : PubMed/NCBI
9	Gur S, Kadowitz PJ and Hellstrom WJ: RhoA/Rho-kinase as a therapeutic target for the male urogenital tract. J Sex Med. 8:675–687. 2011. View Article : Google Scholar
10	Rathinam R, Berrier A and Alahari SK: Role of Rho GTPases and their regulators in cancer progression. Front Biosci (Landmark Ed). 16:2561–2571. 2011. View Article : Google Scholar
11	Kwiatkowska A and Symons M: Signaling determinants of glioma cell invasion. Adv Exp Med Biol. 986:121–141. 2013. View Article : Google Scholar
12	Oh HK, Sin JI, Choi J, Park SH, Lee TS and Choi YS: Overexpression of CD73 in epithelial ovarian carcinoma is associated with better prognosis, lower stage, better differentiation and lower regulatory T cell infiltration. J Gynecol Oncol. 23:274–281. 2012. View Article : Google Scholar : PubMed/NCBI
13	Takami Y, Higashi M, Kumagai S, Kuo PC, Kawana H, Koda K, Miyazaki M and Harigaya K: The activity of RhoA is correlated with lymph node metastasis in human colorectal cancer. Dig Dis Sci. 53:467–473. 2008. View Article : Google Scholar
14	Zhao X, Lu L, Pokhriyal N, Ma H, Duan L, Lin S, Jafari N, Band H and Band V: Overexpression of RhoA induces preneo-plastic transformation of primary mammary epithelial cells. Cancer Res. 69:483–491. 2009. View Article : Google Scholar : PubMed/NCBI
15	Friesland A, Zhao Y, Chen YH, Wang L, Zhou H and Lu Q: Small molecule targeting Cdc42-intersectin interaction disrupts Golgi organization and suppresses cell motility. Proc Natl Acad Sci USA. 110:1261–1266. 2013. View Article : Google Scholar : PubMed/NCBI
16	Li J, Tang LD, Tang WX and Ng SW: Expression and significance of RhoA protein in epithelial ovarian cancer cells. J Endocr Surg. 3:147–153. 2008.In Chinese.
17	Xing L, Yao X, Williams KR and Bassell GJ: Negative regulation of RhoA translation and signaling by hnRNP-Q1 affects cellular morphogenesis. Mol Biol Cell. 23:1500–1509. 2012. View Article : Google Scholar : PubMed/NCBI
18	Shimada T, Nishimura Y, Nishiuma T, Rikitake Y, Hirase T and Yokoyama M: Adenoviral transfer of rho family proteins to lung cancer cells ameliorates cell proliferation and motility and increases apoptotic change. Kobe J Med Sci. 53:125–134. 2007.PubMed/NCBI
19	Wang H, Zhao G, Liu X, Sui A, Yang K, Yao R, Wang Z and Shi Q: Silencing of RhoA and RhoC expression by RNA interference suppresses human colorectal carcinoma growth in vivo. J Exp Clin Cancer Res. 29:1232010. View Article : Google Scholar : PubMed/NCBI
20	Li QQ, Skinner J and Bennett JE: Evaluation of reference genes for real-time quantitative PCR studies in Candida glabrata following azole treatment. BMC Mol Biol. 13:222012. View Article : Google Scholar : PubMed/NCBI
21	Ali HM, Urbinati G, Raouane M and Massaad-Massade L: Significance and applications of nanoparticles in siRNA delivery for cancer therapy. Expert Rev Clin Pharmacol. 5:403–412. 2012. View Article : Google Scholar : PubMed/NCBI
22	Kelly P, Stemmle LN, Madden JF, Fields TA, Daaka Y and Casey PJ: A role for the G12 family of heterotrimeric G proteins in prostate cancer invasion. J Biol Chem. 281:26483–26490. 2006. View Article : Google Scholar : PubMed/NCBI
23	Kelly P, Moeller BJ, Juneja J, Booden MA, Der CJ, Daaka Y, Dewhirst MW, Fields TA and Casey PJ: The G12 family of heterotrimeric G proteins promotes breast cancer invasion and metastasis. Proc Natl Acad Sci USA. 103:8173–8178. 2006. View Article : Google Scholar : PubMed/NCBI
24	Basu Roy UK, Henkhaus RS, Loupakis F, Cremolini C, Gerner EW and Ignatenko NA: Caveolin-1 is a novel regulator of K-RAS-dependent migration in colon carcinogenesis. Int J Cancer. 133:43–57. 2013. View Article : Google Scholar : PubMed/NCBI
25	Nikonova E, Tsyganov MA, Kolch W, Fey D and Kholodenko BN: Control of the G-protein cascade dynamics by GDP dissociation inhibitors. Mol Biosyst. 9:2454–2462. 2013. View Article : Google Scholar : PubMed/NCBI
26	Struckhoff AP, Rana MK and Worthylake RA: RhoA can lead the way in tumor cell invasion and metastasis. Front Biosci (Landmark Ed). 16:1915–1926. 2011. View Article : Google Scholar
27	Fiordalisi JJ, Keller PJ and Cox AD: PRL tyrosine phosphatases regulate rho family GTPases to promote invasion and motility. Cancer Res. 66:3153–3161. 2006. View Article : Google Scholar : PubMed/NCBI
28	Olson MF, Ashworth A and Hall A: An essential role for Rho, Rac, and dc42 GTPases in cell cycle progression through G1. Science. 269:1270–1272. 1995. View Article : Google Scholar : PubMed/NCBI
29	Burridge K and Wennerberg K: Rho and Rac take center stage. Cell. 116:167–179. 2004. View Article : Google Scholar : PubMed/NCBI
30	Gómez del Pulgar T, Benitah SA, Valerón PF, Espina C and Lacal JC: Rho GTPase expression in tumourigenesis: evidence for a significant link. BioEssays. 27:602–613. 2005. View Article : Google Scholar : PubMed/NCBI
31	Qiu RG, Chen J, McCormick F and Symons M: A role for Rho in Ras transformation. Proc Natl Acad Sci USA. 92:11781–11785. 1995. View Article : Google Scholar : PubMed/NCBI
32	Sahai E and Marshall CJ: RHO-GTPases and cancer. Nat Rev Cancer. 2:133–142. 2002. View Article : Google Scholar
33	Chen S, Wang J, Gou WF, Xiu YL, Zheng HC, Zong ZH, Takano Y and Zhao Y: The involvement of RhoA and Wnt-5a in the tumorigenesis and progression of ovarian epithelial carcinoma. Int J Mol Sci. s14:24187–24199. 2013. View Article : Google Scholar
34	Ohta T, Takahashi T, Shibuya T, Amita M, Henmi N, Takahashi K and Kurachi H: Inhibition of the Rho/ROCK pathway enhances the efficacy of cisplatin through the blockage of hypoxia-inducible factor-1α in human ovarian cancer cells. Cancer Biol Ther. 13:25–33. 2012. View Article : Google Scholar : PubMed/NCBI
35	Canet B, Pons C, Espinosa I and Prat J: Ovarian clear cell carcinomas: RHO GTPases may contribute to explain their singular biologic behavior. Hum Pathol. 42:833–839. 2011. View Article : Google Scholar : PubMed/NCBI
36	Gest C, Mirshahi P, Li H, Pritchard LL, Joimel U, Blot E, Chidiac J, Poletto B, Vannier JP, Varin R, et al: Ovarian cancer: Stat3, RhoA and IGF-IR as therapeutic targets. Cancer Lett. 317:207–217. 2012. View Article : Google Scholar
37	Horiuchi A, Kikuchi N, Osada R, Wang C, Hayashi A, Nikaido T and Konishi I: Overexpression of RhoA enhances peritoneal dissemination: RhoA suppression with lovastatin may be useful for ovarian cancer. Cancer Sci. 99:2532–2539. 2008. View Article : Google Scholar : PubMed/NCBI
38	Boudreau RL and Davidson BL: Generation of hairpin-based RNAi vectors for biological and therapeutic application. Methods Enzymol. 507:275–296. 2012. View Article : Google Scholar : PubMed/NCBI
39	Khatri N, Rathi M, Baradia D, Trehan S and Misra A: In vivo delivery aspects of miRNA, shRNA and siRNA. Crit Rev Ther Drug Carrier Syst. 29:487–527. 2012. View Article : Google Scholar : PubMed/NCBI