Knockdown of protein phosphatase 5 inhibits ovarian cancer growth in vitro

Zheng,Xiaojiao; Zhang,Lianxiao; Jin,Bohong; Zhang,Fubin; Zhang,Duoyi; Cui,Lining

doi:10.3892/ol.2015.3828

Knockdown of protein phosphatase 5 inhibits ovarian cancer growth in vitro

Authors:
- Xiaojiao Zheng
- Lianxiao Zhang
- Bohong Jin
- Fubin Zhang
- Duoyi Zhang
- Lining Cui
View Affiliations

Affiliations: Department of Gynaecology and Obstetrics, Ningbo First Hospital, Ningbo, Zhejiang 315010, P.R. China
Published online on: October 26, 2015 https://doi.org/10.3892/ol.2015.3828
Pages: 168-172

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

Ovarian cancer is the most common cause of gynecological cancer-related mortality. Serine/threonine protein phosphatase 5 (PP5, PPP5C) has been recognized to be involved in the regulation of multiple cellular signaling cascades that control diverse cellular processes, including cell growth, differentiation, proliferation, motility and apoptosis. In this study, to evaluate the functional role of PP5 in ovarian cancer cells, lentivirus‑mediated RNA interference (RNAi) was applied to silence PPP5C in the human ovarian cancer cell line CAOV‑3. Cell viability was measured by 3‑(4,5‑dimethylthiazol‑2‑yl)-2,5‑diphenyltetrazolium bromide assay. Cell colony forming ability was measured by colony formation. Cell cycle progression was determined by propidium iodide staining and flow cytometry. The results demonstrated that lentivirus-mediated RNAi specifically suppressed the expression of PPP5C at the mRNA and protein levels in CAOV‑3 cells. Further investigations revealed that PP5 knockdown significantly inhibited the proliferation and colony formation of CAOV‑3 cells. Moreover, the cell cycle of CAOV‑3 cells was arrested at the G0/G1 phase following PP5 knockdown. This study highlights the crucial role of PP5 in promoting ovarian cancer cell proliferation, and provides a foundation for further study into the clinical potential of lentiviral-mediated delivery of PP5 RNAi therapy for the treatment of ovarian cancer.

View Figures

View References

1	Siegel R, Naishadham D and Jemal A: Cancer statistics, 2013. CA Cancer J Clin. 63:11–30. 2013. View Article : Google Scholar : PubMed/NCBI
2	Yu Y, Zhang M, Zhang X, Cai Q, Hong S, Jiang W and Xu C: Synergistic effects of combined platelet-activating factor receptor and epidermal growth factor receptor targeting in ovarian cancer cells. J Hematol Oncol. 7:392014. View Article : Google Scholar : PubMed/NCBI
3	Chalmers MJ, Kolch W, Emmett MR, Marshall AG and Mischak H: Identification and analysis of phosphopeptides. J Chromatogr B Analyt Technol Biomed Life Sci. 803:111–120. 2004. View Article : Google Scholar : PubMed/NCBI
4	Olsen JV, Blagoev B, Gnad F, Macek B, Kumar C, Mortensen P and Mann M: Global, in vivo and site-specific phosphorylation dynamics in signaling networks. Cell. 127:635–648. 2006. View Article : Google Scholar : PubMed/NCBI
5	Ballif BA, Roux PP, Gerber SA, MacKeigan JP, Blenis J and Gygi SP: Quantitative phosphorylation profiling of the ERK/p90 ribosomal S6 kinase-signaling cassette and its targets, the tuberous sclerosis tumor suppressors. Proc Natl Acad Sci USA. 102:667–672. 2005. View Article : Google Scholar : PubMed/NCBI
6	Hoffert JD, Pisitkun T, Wang G, Shen RF and Knepper MA: Quantitative phosphoproteomics of vasopressin-sensitive renal cells: Regulation of aquaporin-2 phosphorylation at two sites. Proc Natl Acad Sci USA. 103:7159–7164. 2006. View Article : Google Scholar : PubMed/NCBI
7	Yang F, Stenoien DL, Strittmatter EF, Wang J, Ding L, Lipton MS, Monroe ME, Nicora CD, Gristenko MA, Tang K, et al: Phosphoproteome profiling of human skin fibroblast cells in response to low- and high-dose irradiation. J Proteome Res. 5:1252–1260. 2006. View Article : Google Scholar : PubMed/NCBI
8	Ham BM, Jayachandran H, Yang F, Jaitly N, Polpitiya AD, Monroe ME, Wang L, Zhao R, Purvine SO, Livesay EA, et al: Novel Ser/Thr protein phosphatase 5 (PP5) regulated targets during DNA damage identified by proteomics analysis. J Proteome Res. 9:945–953. 2010. View Article : Google Scholar : PubMed/NCBI
9	Yamaguchi F, Umeda Y, Shimamoto S, Tsuchiya M, Tokumitsu H, Tokuda M and Kobayashi R: S100 proteins modulate protein phosphatase 5 function: a link between CA2+ signal transduction and protein dephosphorylation. J Biol Chem. 287:13787–13798. 2012. View Article : Google Scholar : PubMed/NCBI
10	Chen MX, McPartlin AE, Brown L, Chen YH, Barker HM and Cohen PT: A novel human protein serine/threonine phosphatase, which possesses four tetratricopeptide repeat motifs and localizes to the nucleus. EMBO J. 13:4278–4290. 1994.PubMed/NCBI
11	Connarn JN, Assimon VA, Reed RA, Tse E, Southworth DR, Zuiderweg ER, Gestwicki JE and Sun D: The molecular chaperone Hsp70 activates protein phosphatase 5 (PP5) by binding the tetratricopeptide repeat (TPR) domain. J Biol Chem. 289:2908–2917. 2014. View Article : Google Scholar : PubMed/NCBI
12	von Kriegsheim A, Pitt A, Grindlay GJ, Kolch W and Dhillon AS: Regulation of the Raf-MEK-ERK pathway by protein phosphatase 5. Nat Cell Biol. 8:1011–1016. 2006. View Article : Google Scholar : PubMed/NCBI
13	Zuo Z, Urban G, Scammell JG, Dean NM, McLean TK, Aragon I and Honkanen RE: Ser/Thr protein phosphatase type 5 (PP5) is a negative regulator of glucocorticoid receptor-mediated growth arrest. Biochemistry. 38:8849–8857. 1999. View Article : Google Scholar : PubMed/NCBI
14	Zuo Z, Dean NM and Honkanen RE: Serine/threonine protein phosphatase type 5 acts upstream of p53 to regulate the induction of p21(WAF1/Cip1) and mediate growth arrest. J Biol Chem. 273:12250–12258. 1998. View Article : Google Scholar : PubMed/NCBI
15	Chinkers M: Targeting of a distinctive protein-serine phosphatase to the protein kinase-like domain of the atrial natriuretic peptide receptor. Proc Natl Acad Sci USA. 91:11075–11079. 1994. View Article : Google Scholar : PubMed/NCBI
16	Silverstein AM, Galigniana MD, Chen MS, Owens-Grillo JK, Chinkers M and Pratt WB: Protein phosphatase 5 is a major component of glucocorticoid receptor hsp90 complexes with properties of an FK506-binding immunophilin. J Biol Chem. 272:16224–16230. 1997. View Article : Google Scholar : PubMed/NCBI
17	Davies TH, Ning YM and Sánchez ER: Differential control of glucocorticoid receptor hormone-binding function by tetratricopeptide repeat (TPR) proteins and the immunosuppressive ligand FK506. Biochemistry. 44:2030–2038. 2005. View Article : Google Scholar : PubMed/NCBI
18	Urban G, Golden T, Aragon IV, Scammell JG, Dean NM and Honkanen RE: Identification of an estrogen-inducible phosphatase (PP5) that converts MCF-7 human breast carcinoma cells into an estrogen-independent phenotype when expressed constitutively. J Biol Chem. 276:27638–27646. 2001. View Article : Google Scholar : PubMed/NCBI
19	Golden T, Aragon IV, Zhou G, Cooper SR, Dean NM and Honkanen RE: Constitutive over expression of serine/threonine protein phosphatase 5 (PP5) augments estrogen-dependent tumor growth in mice. Cancer Lett. 215:95–100. 2004. View Article : Google Scholar : PubMed/NCBI
20	Ali MW, Cacan E, Liu Y, Pierce JY, Creasman WT, Murph MM, Govindarajan R, Eblen ST, Greer SF and Hooks SB: Transcriptional suppression, DNA methylation and histone deacetylation of the regulator of G-protein signaling 10 (RGS10) gene in ovarian cancer cells. PloS one. 8:e601852013. View Article : Google Scholar : PubMed/NCBI
21	Chinkers M: Protein phosphatase 5 in signal transduction. Trends Endocrinol Metab. 12:28–32. 2001. View Article : Google Scholar : PubMed/NCBI
22	Yong W, Bao S, Chen H, Li D, Sánchez ER and Shou W: Mice lacking protein phosphatase 5 are defective in ataxia telangiectasia mutated (ATM)-mediated cell cycle arrest. J Biol Chem. 282:14690–14694. 2007. View Article : Google Scholar : PubMed/NCBI
23	Zhou G, Golden T, Aragon IV and Honkanen RE: Ser/Thr protein phosphatase 5 inactivates hypoxia-induced activation of an apoptosis signal-regulating kinase 1/MKK-4/JNK signaling cascade. J Biol Chem. 279:46595–46605. 2004. View Article : Google Scholar : PubMed/NCBI
24	Huang S, Shu L, Easton J, Harwood FC, Germain GS, Ichijo H and Houghton PJ: Inhibition of mammalian target of rapamycin activates apoptosis signal-regulating kinase 1 signaling by suppressing protein phosphatase 5 activity. J Biol Chem. 279:36490–36496. 2004. View Article : Google Scholar : PubMed/NCBI
25	Urban G, Golden T, Aragon IV, Cowsert L, Cooper SR, Dean NM and Honkanen RE: Identification of a functional link for the p53 tumor suppressor protein in dexamethasone-induced growth suppression. J Biol Chem. 278:9747–9753. 2003. View Article : Google Scholar : PubMed/NCBI
26	Wechsler T, Chen BP, Harper R, Morotomi-Yano K, Huang BC, Meek K, Cleaver JE, Chen DJ and Wabl M: DNA-PKcs function regulated specifically by protein phosphatase 5. Proc Natl Acad Sci USA. 101:1247–1252. 2004. View Article : Google Scholar : PubMed/NCBI