Optimization of Sample Preparation for RT-PCR Gene Expression Analysis during Dose-dependant Drug Toxicity Studies of Xenobiotics Impact on Human Cell Lines

Автор: N.A. Krainova, D.S. Makeeva, D.G. Maksimenko, N.V. Pul'kova, and M.Yu. Shkurnikov

Страница: 71-77

Optimization of Sample Preparation for RT-PCR Gene Expression Analysis during Dose-dependant Drug Toxicity Studies of Xenobiotics Impact on Human Cell Lines

Russian Journal of Biotechnology, 2013, N 6, P. 71-77

UDC 57.081

Section:  “Metrology, Standartization, and Control”

N.A. Krainova ^{1, *},  D.S. Makeeva ^{1, 2},  D.G. Maksimenko ¹,  N.V. Pul'kova ¹,  and  M.Yu. Shkurnikov ³

¹  The Research-and-Technical Center Bioclinicum,  115088, Moscow Russia

²  The Faculty of Bioengineering and Bioinformatics, Lomonosov Moscow State University,  119991, Moscow Russia

³  The Research Institute for General Pathology and Pathophysiology, Russ. Acad. Med. Sci.,  125315, Moscow Russia

e-mail:  n.krainova@bioclinicum.com

An approac to the optimization of sample preparation for the gene expression analysis of a fixed number of marker transcripts in 96-well plates and their analogs during the analysis of toxic effects of drugs within their wide concentration diapason using as a model the impact of cisplatin on of the HMEC-1 microvascular endothelium cell line has been discussed. The most significant changes (up to 10 times) were recorded in the HSP90AA1 gene expression under the equal conditions for all samples preparing. The practical recommendations for the application of the suggested scheme to the analysis of toxicities of other xenobiotics in various in vitro models with a low cell amount were given.

Key words:  cDNA preamplification,  cisplatin,  genes of GSR, FOS, NQO1, FAS, HSP90AA1, HSPA1A, HSPB1 and MYC;  HMEC-1 cell line,  RT-PCR.

31.03.2015, 1458 просмотров.