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Journal

2013 | 11 | 3 | 348-357

Article title

Selection of reference genes for quantitative real-time PCR evaluation of chronic erythropoietin treatment effect on the SH-SY5Y and PC12 cells

Content

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Languages of publication

EN

Abstracts

EN
Abstract The quantitative real-time polymerase chain reaction (qPCR) is a sensitive technique for examining the influence of erythropoietin (Epo) on gene expression. A critical and fundamental step for data analysis is the selection of and normalization to the optimal reference gene(s). We identified appropriate reference gene(s) among 32 genes during chronic recombinant human Epo (rHuEpo) treatment of SH-SY5Y cells using TaqMan human Express Endogenous Control Plate. Expression stability of the selected reference gene (RPLP) was retested with qPCR, together with two commonly used reference genes (GAPDH, ACTB) and six genes of interest (EPOR, EPO, STAT5B, STAT5A, JUN, AKT). In PC12 cells, three commonly used reference genes (Gapdh, CycA and Ywhaz) and seven genes of interest (EpoR, Epo, Stat5b, Stat5a, Jun, Akt, Fos) were evaluated. For the evaluation of expression stability, geNorm, NormFinder and BestKeeper software were used. All three gave similar results. We demonstrated that among the housekeeping genes, RPLP in SH-SY5Y and CycA and Ywhaz in PC12 are the most stable genes. Additionally, we showed that normalization with GAPDH gave misleading results compared to normalization with geNorm. In conclusion, selection of the appropriate normalization gene(s) is crucial for correct interpretation of rHuEpo treatment results. Graphical abstract [...]

Publisher

Journal

Year

Volume

11

Issue

3

Pages

348-357

Physical description

Dates

published
1 - 3 - 2013
online
28 - 12 - 2012

References

  • [1] E.B. Thorling, A.J. Erslev, Blood 31, 332 (1968)
  • [2] H.F. Bunn, Blood 109, 868 (2007) http://dx.doi.org/10.1182/blood-2006-08-019083[Crossref]
  • [3] N. Debeljak, A.J. Sytkowski. EPOR. UCSD-Nature Molecule pages. 2007. DOI: 10.1038/mp.a000863.01
  • [4] C. Chen, A.J. Sytkowski, Blood 104, 73 (2004) http://dx.doi.org/10.1182/blood-2003-04-1340[Crossref]
  • [5] A.J. Sytkowski, Erythropoietin: Blood, Brain and Beyond (Wiley-VCH, Weinheim, 2004) http://dx.doi.org/10.1002/3527602380[Crossref]
  • [6] H.H. Marti, et al., Eur. J. Neurosci. 8, 666 (1996) http://dx.doi.org/10.1111/j.1460-9568.1996.tb01252.x[Crossref]
  • [7] T. Kobayashi, H. Yanase, T. Iwanaga, R. Sasaki, M. Nagao, Biochem. Biophys. Res. Commun. 296, 145 (2002) http://dx.doi.org/10.1016/S0006-291X(02)00832-X[Crossref]
  • [8] Z. Cai, et al., Circulation 108, 79 (2003) http://dx.doi.org/10.1161/01.CIR.0000078635.89229.8A[Crossref]
  • [9] M.L. Brines, et al., Proceedings of the National Academy of Sciences of the United States of America 97, 10526 (2000) http://dx.doi.org/10.1073/pnas.97.19.10526[Crossref]
  • [10] M.A. Bogoyevitch, Cardiovasc. Res. 63, 208 (2004) http://dx.doi.org/10.1016/j.cardiores.2004.03.017[Crossref]
  • [11] G. Grasso, A. Sfacteria, A. Cerami, M. Brines, Neuroscientist 10, 93 (2004) http://dx.doi.org/10.1177/1073858403259187[Crossref]
  • [12] G. Grasso, et al., J. Neurosurg. Spine 4, 310 (2006) http://dx.doi.org/10.3171/spi.2006.4.4.310[Crossref]
  • [13] A.J. Sytkowski, Cell Mol. Neurobiol. 6, 931 (2011) http://dx.doi.org/10.1007/s10571-011-9695-0[Crossref]
  • [14] W. Feng, H. Wei, G.T. Liu, Mol. Neurobiol. 31, 295 (2005) http://dx.doi.org/10.1385/MN:31:1-3:295[Crossref]
  • [15] H.R. Xie, L.S. Hu, G.Y. Li, Chin. Med. J. (Engl) 123, 1086 (2010)
  • [16] S.A. Bustin, V. Benes, T. Nolan, M.W. Pfaffl, J. Mol. Endocrinol. 34, 597 (2005) http://dx.doi.org/10.1677/jme.1.01755[Crossref]
  • [17] J. Huggett, K. Dheda, S. Bustin, A. Zumla, Genes Immun. 6, 279 (2005) http://dx.doi.org/10.1038/sj.gene.6364190[Crossref]
  • [18] O. Thellin, et al., J. Biotechnol. 75, 291 (1999) http://dx.doi.org/10.1016/S0168-1656(99)00163-7[Crossref]
  • [19] F. Ohl, et al., J. Mol. Med. 83, 1014 (2005) http://dx.doi.org/10.1007/s00109-005-0703-z[Crossref]
  • [20] B.E. Bonefeld, B. Elfving, G. Wegener, Synapse 62, 302 (2008) http://dx.doi.org/10.1002/syn.20496[Crossref]
  • [21] S.A. Bustin, J. Mol. Endocrinol. 25, 169 (2000) http://dx.doi.org/10.1677/jme.0.0250169[Crossref]
  • [22] T. Suzuki, P.J. Higgins, D.R. Crawford, Biotechniques 29, 332 (2000)
  • [23] J.A. Warrington, A. Nair, M. Mahadevappa, M. Tsyganskaya, Physiol. Genomics 2, 143 (2000)
  • [24] H.K. Hamalainen, et al., Anal. Biochem. 299, 63 (2001) http://dx.doi.org/10.1006/abio.2001.5369[Crossref]
  • [25] A. Radonic, et al., Virol. J. 2, 7 (2005) http://dx.doi.org/10.1186/1743-422X-2-7[Crossref]
  • [26] M. Jung, et al., BMC Mol. Biol. 8, 47 (2007) http://dx.doi.org/10.1186/1471-2199-8-47[Crossref]
  • [27] S. Waxman, E. Wurmbach, BMC Genomics. 8, 243 (2007) http://dx.doi.org/10.1186/1471-2164-8-243[Crossref]
  • [28] J. Vandesompele, et al., Genome Biol. 3, RESEARCH0034 (2002)
  • [29] C.L. Andersen, J.L. Jensen, T.F. Orntoft, Cancer Res. 64, 5245 (2004) http://dx.doi.org/10.1158/0008-5472.CAN-04-0496[Crossref]
  • [30] M.W. Pfaffl, A. Tichopad, C. Prgomet, T.P. Neuvians, Biotechnol. Lett. 26, 509 (2004) http://dx.doi.org/10.1023/B:BILE.0000019559.84305.47[Crossref]
  • [31] B.R. Everaert, G.A. Boulet, J.P. Timmermans, C.J. Vrints, PLoS One 6, e23793 (2011) http://dx.doi.org/10.1371/journal.pone.0023793[Crossref]
  • [32] T. Fink, et al., BMC Mol. Biol. 9, 98 (2008) http://dx.doi.org/10.1186/1471-2199-9-98[Crossref]
  • [33] L. Zhou, Q.E. Lim, G. Wan, H.P. Too, BMC Genomics 11, 75 (2010) http://dx.doi.org/10.1186/1471-2164-11-75[Crossref]
  • [34] M.J. Renzi, et al., Brain Res. Mol. Brain. Res. 104, 86 (2002) http://dx.doi.org/10.1016/S0169-328X(02)00323-6[Crossref]
  • [35] X. Liu, J. Shen, Y. Jin, M. Duan, J. Xu, Indian J. Med. Res. 124, 343 (2006)
  • [36] M. Um, A.W. Gross, H.F. Lodish, Cell Signal 19, 634 (2007) http://dx.doi.org/10.1016/j.cellsig.2006.08.014[Crossref]
  • [37] Z.B. Liao, et al., Neurol. India 57, 722 (2009) http://dx.doi.org/10.4103/0028-3886.59466[Crossref]

Document Type

Publication order reference

Identifiers

YADDA identifier

bwmeta1.element.-psjd-doi-10_2478_s11532-012-0164-5
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