Int. J. Mol. Sci. 2012, 13(3), 2810-2826; doi:10.3390/ijms13032810
Article

Variation in Stability of Endogenous Reference Genes in Fallopian Tubes and Endometrium from Healthy and Ectopic Pregnant Women

1email, 2email, 1email, 1email and 1,* email
Received: 5 January 2012; in revised form: 10 February 2012 / Accepted: 27 February 2012 / Published: 2 March 2012
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract: RT-qPCR is commonly employed in gene expression studies in ectopic pregnancy. Most use RN18S1, β-actin or GAPDH as internal controls without validation of their suitability as reference genes. A systematic study of the suitability of endogenous reference genes for gene expression studies in ectopic pregnancy is lacking. The aims of this study were therefore to evaluate the stability of 12 reference genes and suggest those that are stable for use as internal control genes in fallopian tubes and endometrium from ectopic pregnancy and healthy non-pregnant controls. Analysis of the results showed that the genes consistently ranked in the top six by geNorm and NormFinder algorithms, were UBC, GAPDH, CYC1 and EIF4A2 (fallopian tubes) and UBC and ATP5B (endometrium). mRNA expression of NAPE-PLD as a test gene of interest varied between the groups depending on which of the 12 reference genes was used as internal controls. This study demonstrates that arbitrary selection of reference genes for normalisation in RT-qPCR studies in ectopic pregnancy without validation, risk producing inaccurate data and should therefore be discouraged.
Keywords: ectopic pregnancy; fallopian tubes; endometrium; reference genes; geNorm; NormFinder; quantitative real time PCR; NAPE-PLD
PDF Full-text Download PDF Full-Text [574 KB, uploaded 19 June 2014 04:12 CEST]

Export to BibTeX |
EndNote


MDPI and ACS Style

Gebeh, A.K.; Marczylo, E.L.; Amoako, A.A.; Willets, J.M.; Konje, J.C. Variation in Stability of Endogenous Reference Genes in Fallopian Tubes and Endometrium from Healthy and Ectopic Pregnant Women. Int. J. Mol. Sci. 2012, 13, 2810-2826.

AMA Style

Gebeh AK, Marczylo EL, Amoako AA, Willets JM, Konje JC. Variation in Stability of Endogenous Reference Genes in Fallopian Tubes and Endometrium from Healthy and Ectopic Pregnant Women. International Journal of Molecular Sciences. 2012; 13(3):2810-2826.

Chicago/Turabian Style

Gebeh, Alpha K.; Marczylo, Emma L.; Amoako, Akwasi A.; Willets, Jonathon M.; Konje, Justin C. 2012. "Variation in Stability of Endogenous Reference Genes in Fallopian Tubes and Endometrium from Healthy and Ectopic Pregnant Women." Int. J. Mol. Sci. 13, no. 3: 2810-2826.

Int. J. Mol. Sci. EISSN 1422-0067 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert