Reference Genes for Quantitative RT-PCR Studies on Caco-2 Cell Differentiation

Автор: E.V. Trushkin, D.G. Maksimenko, N.A. Khaustova, I.N. Nechaev, N.G. Luzgina, D.V. Maltseva, and A.L. Rusanov

Страница: 33-41

Reference Genes for Quantitative RT-PCR Studies on Caco-2 Cell Differentiation

Biotekhnologiya, 2013, N 1, P. 33-41

UDC 57.04, 57.041, 57.032

Section:  “Producers, Biology, Selection, and Gene Engineering”

E.V. Trushkin ¹,  D.G. Maksimenko ¹,  N.A. Khaustova ¹,  I.N. Nechaev ²,  N.G. Luzgina ³,  D.V. Maltseva ¹,  and  A.L. Rusanov ^{1, *}

¹  The Research and Technique Center BioClinicum,  115088, Moscow Russia

²  The Institute of General Pathology and Pathophysiology, Russ. Acad. Med. Sci.,  125315, Moscow Russia

³  The Research Center for Clinical and Experimental Medicine, Russ. Acad. Med. Sci., Sibirian Branch,  630117, Novosibirsk Russia

e-mail:  a.rusanov@bioclinicum.com

Changing of the expression of 16 putative reference genes (ACTB, GAPDH, GUS, RPLPO,TBP, PSMC4, PUM1, MRPL19, SF3A1, HPRT1, RPS18,UBC, B2M, HSPC3, SDHA, YWHAZ) in a Caco-2 cell line before and after differentiation has been assessed. The expression stability was determined using two methods, ΔCt and geNorm. It was shown that SF3A1, SDHA and TBP genes are expressed more constantly than ACTB and GAPDH that conventionally serve as reference. A quantitative assessment of the changing in SI, SLC11A2, MYO1A, ANPEP  and PPARG gene expression (characteristic genes of small gut) confirmed that  SF3A1, SDHA and TBP are more reliable as reference genes. The present investigation can provide a methodical basis for the study in detail of the Caco-2 cell differentiation by RT-PCR.

Key words:  Caco-2 cell line,  differentiated cell lines,  expression stability,  real-time PCR,  reference genes.

The full English version of the article was published in “Biotechnology in Russia”, 2013, Issue 1, pp. 33-41 as E.V. Trushkin, D.G. Maksimenko, N.A. Khaustova, I.N. Nechaev, N.G. Luzgina, D.V. Mal’tseva, and A.L. Rusanov “Reference Genes for Quantitative RT-PCR Studies on Caco-2 Cell Differentiation”.

It is contained at the Russian Scientific Electron Library website:  http://elibrary.ru/item.asp?id=21987104

27.04.2015, 1812 просмотров.