PML silencing inhibits cell proliferation and induces DNA damage in cultured ovarian cancer cells

Liu,Sheng‑Bing; Shen,Zhong‑Fei; Guo,Yan‑Jun; Cao,Li‑Xian; Xu,Ying

doi:10.3892/br.2017.919

PML silencing inhibits cell proliferation and induces DNA damage in cultured ovarian cancer cells

Authors:
- Sheng‑Bing Liu
- Zhong‑Fei Shen
- Yan‑Jun Guo
- Li‑Xian Cao
- Ying Xu
View Affiliations

Affiliations: College of Medicine, Jiaxing University, Jiaxing, Zhejiang 314001, P.R. China
Published online on: May 24, 2017 https://doi.org/10.3892/br.2017.919
Pages: 29-35
Copyright: © Liu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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 promyelocytic leukemia (PML) gene is a tumor suppressor gene. It was first identified in acute promyelocytic leukemia, in which it is fused to retinoic acid receptor α by the (15;17) chromosomal translocation. The function of the PML protein is frequently lost or aberrant in human solid tumors. In human ovarian carcinoma tissue, PML detected by immunohistochemistry was highly expressed. A PML‑silencing vector, pSRG‑shPml, was constructed and used to transfect human ovarian cancer cells. Cells were cultured and selected with puromycin for 10‑15 days, and then the PML mRNA expression levels were detected by RT-qPCR and immunofluorescence. Proliferation and clone number of PML‑depleted cells were detected using MTT assay and colony‑forming assay. The protein expression associated with DNA damage and apoptosis was assessed in PML‑depleted cells using western blot analysis and immunofluorescence. The results showed that PML was highly expressed in human ovarian tissue. The proliferation and colony formation of ovarian cancer cells were significantly inhibited after PML was depleted. Western blot analysis and immunofluorescence revealed that p‑H2AX and cleaved caspase‑3 expression significantly increased after PML silencing. PML was located in the nucleus, and it formed foci after X‑ray irradiation. PML foci increased significantly with increasing irradiation doses.

View Figures

View References

1	Dutrieux J, Maarifi G, Portilho DM, Arhel NJ, Chelbi-Alix MK and Nisole S: PML/TRIM19-dependent inhibition of retroviral reverse-transcription by Daxx. PLoS Pathog. 11:e10052802015. View Article : Google Scholar : PubMed/NCBI
2	Fagioli M, Alcalay M, Pandolfi PP, Venturini L, Mencarelli A, Simeone A, Acampora D, Grignani F and Pelicci PG: Alternative splicing of PML transcripts predicts coexpression of several carboxy-terminally different protein isoforms. Oncogene. 7:1083–1091. 1992.PubMed/NCBI
3	Sahin U, de Thé H and Lallemand-Breitenbach V: PML nuclear bodies: assembly and oxidative stress-sensitive sumoylation. Nucleus. 5:499–507. 2014. View Article : Google Scholar : PubMed/NCBI
4	Bernardi R, Papa A and Pandolfi PP: Regulation of apoptosis by PML and the PML-NBs. Oncogene. 27:6299–6312. 2008. View Article : Google Scholar : PubMed/NCBI
5	Collins SJ: Retinoic acid receptors, hematopoiesis and leukemogenesis. Curr Opin Hematol. 15:346–351. 2008. View Article : Google Scholar : PubMed/NCBI
6	Jensen K, Shiels C and Freemont PS: PML protein isoforms and the RBCC/TRIM motif. Oncogene. 20:7223–7233. 2001. View Article : Google Scholar : PubMed/NCBI
7	Shen TH, Lin HK, Scaglioni PP, Yung TM and Pandolfi PP: The mechanisms of PML-nuclear body formation. Mol Cell. 24:331–339. 2006. View Article : Google Scholar : PubMed/NCBI
8	Duprez E, Saurin AJ, Desterro JM, Lallemand-Breitenbach V, Howe K, Boddy MN, Solomon E, de Thé H, Hay RT and Freemont PS: SUMO-1 modification of the acute promyelocytic leukaemia protein PML: Implications for nuclear localisation. J Cell Sci. 112:381–393. 1999.PubMed/NCBI
9	Zhong S, Müller S, Ronchetti S, Freemont PS, Dejean A and Pandolfi PP: Role of SUMO-1-modified PML in nuclear body formation. Blood. 95:2748–2752. 2000.PubMed/NCBI
10	Bernardi R and Pandolfi PP: Structure, dynamics and functions of promyelocytic leukaemia nuclear bodies. Nat Rev Mol Cell Biol. 8:1006–1016. 2007. View Article : Google Scholar : PubMed/NCBI
11	Wethkamp N and Klempnauer KH: Daxx is a transcriptional repressor of CCAAT/enhancer-binding protein β. J Biol Chem. 284:28783–28794. 2009. View Article : Google Scholar : PubMed/NCBI
12	Hofmann TG and Will H: Body language: The function of PML nuclear bodies in apoptosis regulation. Cell Death Differ. 10:1290–1299. 2003. View Article : Google Scholar : PubMed/NCBI
13	Pan WW, Zhou JJ, Liu XM, Xu Y, Guo LJ, Yu C, Shi QH and Fan HY: Death domain-associated protein DAXX promotes ovarian cancer development and chemoresistance. J Biol Chem. 288:13620–13630. 2013. View Article : Google Scholar : PubMed/NCBI
14	Salomoni P, Dvorkina M and Michod D: Role of the promyelocytic leukaemia protein in cell death regulation. Cell Death Dis. 3:e2472012. View Article : Google Scholar : PubMed/NCBI
15	van Damme E, Laukens K, Dang TH and Van Ostade X: A manually curated network of the PML nuclear body interactome reveals an important role for PML-NBs in SUMOylation dynamics. Int J Biol Sci. 6:51–67. 2010. View Article : Google Scholar : PubMed/NCBI
16	Chen RH, Lee YR and Yuan WC: The role of PML ubiquitination in human malignancies. J Biomed Sci. 19:812012. View Article : Google Scholar : PubMed/NCBI
17	Chen RH, Lee YR and Yuan WC: The role of PML ubiquitination in human malignancies. J Biomed Sci. 19:812012. View Article : Google Scholar : PubMed/NCBI
18	Guo A, Salomoni P, Luo J, Shih A, Zhong S, Gu W and Pandolfi PP: The function of PML in p53-dependent apoptosis. Nat Cell Biol. 2:730–736. 2000. View Article : Google Scholar : PubMed/NCBI
19	Wang ZG, Ruggero D, Ronchetti S, Zhong S, Gaboli M, Rivi R and Pandolfi PP: PML is essential for multiple apoptotic pathways. Nat Genet. 20:266–272. 1998. View Article : Google Scholar : PubMed/NCBI
20	Pearson M, Carbone R, Sebastiani C, Cioce M, Fagioli M, Saito S, Higashimoto Y, Appella E, Minucci S, Pandolfi PP, et al: PML regulates p53 acetylation and premature senescence induced by oncogenic Ras. Nature. 406:207–210. 2000. View Article : Google Scholar : PubMed/NCBI
21	Martin N, Benhamed M, Nacerddine K, Demarque MD, van Lohuizen M, Dejean A and Bischof O: Physical and functional interaction between PML and TBX2 in the establishment of cellular senescence. EMBO J. 31:95–109. 2012. View Article : Google Scholar : PubMed/NCBI
22	Chen HY, Hu JY, Chen TH, Lin YC, Liu X, Lin MY, Lang YD, Yen Y and Chen RH: KLHL39 suppresses colon cancer metastasis by blocking KLHL20-mediated PML and DAPK ubiquitination. Oncogene. 34:5141–5151. 2015. View Article : Google Scholar : PubMed/NCBI
23	Hsu KS, Guan BJ, Cheng X, Guan D, Lam M, Hatzoglou M and Kao HY: Translational control of PML contributes to TNFα-induced apoptosis of MCF7 breast cancer cells and decreased angiogenesis in HUVECs. Cell Death Differ. 23:469–483. 2016. View Article : Google Scholar : PubMed/NCBI
24	Ito K, Carracedo A, Weiss D, Arai F, Ala U, Avigan DE, Schafer ZT, Evans RM, Suda T, Lee CH, et al: A PML-PPAR-δ pathway for fatty acid oxidation regulates hematopoietic stem cell maintenance. Nat Med. 18:1350–1358. 2012. View Article : Google Scholar : PubMed/NCBI
25	Legartová S, Sehnalová P, Malyšková B, Küntziger T, Collas P, Cmarko D, Raška I, Sorokin DV, Kozubek S and Bártová E: Localized movement and levels of 53BP1protein are changed by γ-irradiation in PML deficient cells. J Cell Biochem. 117:2583–2596. 2016. View Article : Google Scholar : PubMed/NCBI
26	Van Colombo Ν, Gorp T, Parma G, Amant F, Gatta G, Sessa C and Vergote I: Ovarian cancer. Crit Rev Oncol Hematol. 60:159–179. View Article : Google Scholar : PubMed/NCBI
27	Siegel RL, Miller KD and Jemal A: Cancer statistics, 2015. CA Cancer J Clin. 65:5–29. 2015. View Article : Google Scholar : PubMed/NCBI