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Medical Oncology

, 34:27 | Cite as

Knockdown of PYCR1 inhibits cell proliferation and colony formation via cell cycle arrest and apoptosis in prostate cancer

  • Tengyue Zeng
  • Libing Zhu
  • Min Liao
  • Wenli Zhuo
  • Shunliang Yang
  • Weizhen Wu
  • Dong WangEmail author
Original Paper

Abstract

Pyrroline-5-carboxylate reductase 1 (PYCR1) is an enzyme involved in cell metabolism, which has been shown to be up-regulated in cancers. However, the functions of PYCR1 in prostate cancers (PCa) are still largely unknown. In the present study, we found that PYCR1 was highly expressed in prostate cancer tissues and then knocked down PYCR1 in PCa cell lines (DU145, PC-3 and LNCap) via lentivirus-mediated gene delivery and analyzed its biological function. Both qRT-PCR and western blotting indicated that PYCR1 was suppressed efficiently after sh-PYCR1 infection. Further analysis indicated knockdown of PYCR1 significantly inhibited PCa cell growth and colony formation ability. The inhibition effects on growth were likely due to G2/M-phase arrest and enhanced cell apoptosis, as determined by flow cytometer analysis. At last, we verified the expression levels of cell cycle regulatory proteins, including CDK1, CDK2, CDK4 and Cyclin B1 were all downregulated and cell apoptotic-related proteins, including cleaved caspase 3 and cleaved PARP were increased in PCa cells after PYCR1 knockdown. Furthermore, PYCR1 has been shown not to be directly regulated by androgen receptor (AR) levels. These results show the functions of PYCR1 in PCa tumorigenesis for the first time and suggest that PYCR1 might be a good potential therapy approach for treating PCa.

Keywords

Prostate cancer PYCR1 Cell proliferation Cell cycle Apoptosis AR signaling 

Notes

Acknowledgements

The authors are thankful for the financial support from the National Natural Science Foundation of China (81272247 and 81372751).

Compliance with ethical standards

Conflict of interest

Conflict of interest relevant to this article was not reported.

Informed consent

Informed consent was obtained from all individual participants included in the study.

Ethical approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional of Fuzhou General Hospital Affiliated to Fujian Medical University.

Supplementary material

12032_2016_870_MOESM1_ESM.tif (1.3 mb)
Figure S1. (A) The gene delivery efficiency of shPYCR1(S1) in LNCap cells. Upper panels, bright field; lower panels, GFP fluorescence (green). Scale bar, 10 μm. (B) Western blotting analysis of PYCR1 protein levels in shPYCR1(S1)-infected LNCap cells. (C) The proliferation levels of LNCap cells after shPYCR1(S1) infection analyzed by the MTT assay. Data are expressed as mean ± standard deviation (SD) of three independent experiments. ***p < 0.001. (D) Western blot analysis of PYCR1 and PSA protein levels in Con, AR inhibitor (Bicalutamide) or AR activator (DHT) treated LNCap cells. (TIFF 1329 kb)

References

  1. 1.
    Cancer statistics. JAMA. 2013;310:982. doi: 10.1001/jama.2013.5289.
  2. 2.
    Ferlay J, Steliarova-Foucher E, Lortet-Tieulent J, Rosso S, Coebergh JW, Comber H, et al. Cancer incidence and mortality patterns in Europe: estimates for 40 countries in 2012. Eur J Cancer. 2013;49:1374–403.CrossRefPubMedGoogle Scholar
  3. 3.
    Baade PD, Youlden DR, Cramb SM, Dunn J, Gardiner RA. Epidemiology of prostate cancer in the Asia-Pacific region. Prostate Int. 2013;1:47–58.CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Scher HI, Buchanan G, Gerald W, Butler LM, Tilley WD. Targeting the androgen receptor: improving outcomes for castration-resistant prostate cancer. Endocr Relat Cancer. 2004;11:459–76.CrossRefPubMedGoogle Scholar
  5. 5.
    Gomella LG, Petrylak DP, Shayegan B. Current management of advanced and castration resistant prostate cancer. Can J Urol. 2014;21:1–6.PubMedGoogle Scholar
  6. 6.
    Struys EA, Jansen EE, Salomons GS. Human pyrroline-5-carboxylate reductase (PYCR1) acts on Delta(1)-piperideine-6-carboxylate generating L-pipecolic acid. J Inherit Metab Dis. 2014;37:327–32.CrossRefPubMedGoogle Scholar
  7. 7.
    Guernsey DL, Jiang H, Evans SC, Ferguson M, Matsuoka M, Nightingale M, et al. Mutation in pyrroline-5-carboxylate reductase 1 gene in families with cutis laxa type 2. Am J Hum Genet. 2009;85:120–9.CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Reversade B, Escande-Beillard N, Dimopoulou A, Fischer B, Chng SC, Li Y, et al. Mutations in PYCR1 cause cutis laxa with progeroid features. Nat Genet. 2009;41:1016–21.CrossRefPubMedGoogle Scholar
  9. 9.
    Kretz R, Bozorgmehr B, Kariminejad MH, Rohrbach M, Hausser I, Baumer A, et al. Defect in proline synthesis: pyrroline-5-carboxylate reductase 1 deficiency leads to a complex clinical phenotype with collagen and elastin abnormalities. J Inherit Metab Dis. 2011;34:731–9.CrossRefPubMedGoogle Scholar
  10. 10.
    Lin DS, Yeung CY, Liu HL, Ho CS, Shu CH, Chuang CK, et al. A novel mutation in PYCR1 causes an autosomal recessive cutis laxa with premature aging features in a family. Am J Med Genet Part A. 2011;155A:1285–9.CrossRefPubMedGoogle Scholar
  11. 11.
    Dimopoulou A, Fischer B, Gardeitchik T, Schroter P, Kayserili H, Schlack C, et al. Genotype-phenotype spectrum of PYCR1-related autosomal recessive cutis laxa. Mol Genet Metab. 2013;110:352–61.CrossRefPubMedGoogle Scholar
  12. 12.
    Scherrer DZ, Baptista MB, Matos AH, Maurer-Morelli CV, Steiner CE. Mutations in PYCR1 gene in three families with autosomal recessive cutis laxa, type 2. Eur J Med Genet. 2013;56:336–9.CrossRefPubMedGoogle Scholar
  13. 13.
    Ernst T, Hergenhahn M, Kenzelmann M, Cohen CD, Bonrouhi M, Weninger A, et al. Decrease and gain of gene expression are equally discriminatory markers for prostate carcinoma: a gene expression analysis on total and microdissected prostate tissue. Am J Pathol. 2002;160:2169–80.CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Jariwala U, Prescott J, Jia L, Barski A, Pregizer S, Cogan JP, et al. Identification of novel androgen receptor target genes in prostate cancer. Mol Cancer. 2007;6:39.CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Maxwell SA, Davis GE. Differential gene expression in p53-mediated apoptosis-resistant vs. apoptosis-sensitive tumor cell lines. Proc Natl Acad Sci USA. 2000;97:13009–14.CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Krishnan N, Dickman MB, Becker DF. Proline modulates the intracellular redox environment and protects mammalian cells against oxidative stress. Free Radic Biol Med. 2008;44:671–81.CrossRefPubMedGoogle Scholar
  17. 17.
    Yasuda T, Kaji Y, Agatsuma T, Niki T, Arisawa M, Shuto S, et al. DJ-1 cooperates with PYCR1 in cell protection against oxidative stress. Biochem Biophys Res Commun. 2013;436:289–94.CrossRefPubMedGoogle Scholar
  18. 18.
    Liu W, Le A, Hancock C, Lane AN, Dang CV, Fan TW, et al. Reprogramming of proline and glutamine metabolism contributes to the proliferative and metabolic responses regulated by oncogenic transcription factor c-MYC. Proc Natl Acad Sci USA. 2012;109:8983–8.CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Marker PC, Donjacour AA, Dahiya R, Cunha GR. Hormonal, cellular, and molecular control of prostatic development. Dev Biol. 2003;253:165–74.CrossRefPubMedGoogle Scholar
  20. 20.
    Geck P, Szelei J, Jimenez J, Lin TM, Sonnenschein C, Soto AM. Expression of novel genes linked to the androgen-induced, proliferative shutoff in prostate cancer cells. J Steroid Biochem Mol Biol. 1997;63:211–8.CrossRefPubMedGoogle Scholar
  21. 21.
    Bonkhoff H, Remberger K. Differentiation pathways and histogenetic aspects of normal and abnormal prostatic growth: a stem cell model. Prostate. 1996;28:98–106.CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  • Tengyue Zeng
    • 1
  • Libing Zhu
    • 2
  • Min Liao
    • 1
  • Wenli Zhuo
    • 1
  • Shunliang Yang
    • 1
  • Weizhen Wu
    • 1
  • Dong Wang
    • 1
    Email author
  1. 1.Department of Urology, Fuzhou General HospitalFujian Medical UniversityFuzhouChina
  2. 2.Department of UrologyLushan Sanatorium of the PLALushanChina

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