caa a22 4500 44582297X CHVBK 20180317145250.0 cr unu---uuuuu 170323e20111201xx s 000 0 eng 10.1007/s10616-011-9383-4 doi (NATIONALLICENCE)springer-10.1007/s10616-011-9383-4 UBC and YWHAZ as suitable reference genes for accurate normalisation of gene expression using MCF7, HCT116 and HepG2 cell lines [Elektronische Daten] [Siang Chua, Wei See Too, Boon Khoo, Ling Few] Relative quantification of in vitro gene expression using real-time PCR requires stably expressed reference gene for normalisation. In this study, total RNA from MCF7, HCT116 and HepG2 cells were extracted and converted to cDNA using commercially available kit, and real-time PCR was then performed to analyse the expression levels of twelve reference genes to select the most ideal reference gene for accurate normalisation in gene expression study. geNorm and NormFinder software were used to analyse the stabilities of the reference genes, which showed a wide range of Ct values. The geNorm analysis showed the following ranking for stability of genes: UBC, YWHAZ>RPLP>TBP>ACTB>HPRT1>PPIA>GAPDH>GUSB>B2M>TUBB>RRN18S. A similar ranking of reference genes was obtained by NormFinder, and the four most stable reference genes were identical using both approaches. UBC and YWHAZ were proposed to be the two most suitable reference genes based on the above analyses. To further assess the stabilities of the UBC and YWHAZ in a formal experiment, MCF7, HCT116 and HepG2 cell lines were subjected to treatments with 5-aza-dC and TSA. Both UBC and YWHAZ exhibited stable expression levels across control and treatment groups. Therefore, we propose that UBC and YWHAZ are the two most suitable reference genes for our gene expression studies using MCF7, HCT116 and HepG2 cell lines. Springer Science+Business Media B.V., 2011 Gene expression nationallicence Normalisation nationallicence Reference gene nationallicence Expression level nationallicence Expression stability nationallicence Real-time PCR nationallicence Chua Siang School of Health Sciences, Universiti Sains Malaysia, 16150, Kubang Kerian, Kelantan, Malaysia aut See Too Wei School of Health Sciences, Universiti Sains Malaysia, 16150, Kubang Kerian, Kelantan, Malaysia aut Khoo Boon Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, 11800, USM, Penang, Malaysia aut Few Ling School of Health Sciences, Universiti Sains Malaysia, 16150, Kubang Kerian, Kelantan, Malaysia aut Cytotechnology Springer Netherlands 63/6(2011-12-01), 645-654 0920-9069 63:6<645 2011 63 10616 https://doi.org/10.1007/s10616-011-9383-4 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s10616-011-9383-4 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Chua Siang School of Health Sciences, Universiti Sains Malaysia, 16150, Kubang Kerian, Kelantan, Malaysia aut NATIONALLICENCE 700 1- See Too Wei School of Health Sciences, Universiti Sains Malaysia, 16150, Kubang Kerian, Kelantan, Malaysia aut NATIONALLICENCE 700 1- Khoo Boon Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, 11800, USM, Penang, Malaysia aut NATIONALLICENCE 700 1- Few Ling School of Health Sciences, Universiti Sains Malaysia, 16150, Kubang Kerian, Kelantan, Malaysia aut NATIONALLICENCE 773 0- Cytotechnology Springer Netherlands 63/6(2011-12-01), 645-654 0920-9069 63:6<645 2011 63 10616 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer