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Variation in stability of housekeeping genes in healthy and adhesion-related mesothelium

Sadek, Khaled H., Cagampang, F.R., Bruce, K.D., Macklon, N. and Cheong, Ying (2012) Variation in stability of housekeeping genes in healthy and adhesion-related mesothelium Fertility and Sterility, 98, (4), pp. 1023-1027. (doi:10.1016/j.fertnstert.2012.06.033). (PMID:22795637).

Record type: Article

Abstract

Objective
To investigate the stability of various housekeeping genes (HKG) within healthy versus scarred peritoneal mesothelium. The use of HKG as internal controls for quantitative real-time polymerase chain reaction (qRT-PCR) studies is based on the assumption of their inherent stability. However, recent evidence suggests that this is not true for all HKG and that stability may be tissue specific and affected by certain pathologies.

Design
Paired mesothelial (n = 10) and adhesion tissue samples (n = 10) were taken during laparoscopic surgery. The stability of 12 candidate reference genes in the mesothelial tissues were evaluated; these include ATP5b, SDHA, CYC1, 18S rRNA, RPL13A, ACTB, YWHAZ, TOP1, UBC, EIF4A2, GAPDH, and B2M.

Setting
Hospital.

Patient(s)
Female patients undergoing laparoscopic gynecological surgery were recruited from the Princess Anne Hospital, United Kingdom.

Intervention(s)
Assessment of HKG expression stability using geNorm algorithm software.

Main Outcome Measure(s)
Stability measure (M) generated by geometric averaging of multiple target genes and mean pairwise variation of genes.

Result(s)
The most stable HKGs observed across both healthy and adhesion-related mesothelium were found to be ACTB, YWHAZ, and CYC1. ACTB had a higher expression in healthy mesothelium compared with in peritoneal adhesion tissue.

Conclusion(s)
This study indicates that ACTB, YWHAZ, and CYC1 are the appropriate internal controls for qRT-PCR analysis in mesothelial gene expression studies. Published discrepancies in gene expression studies using the mesothelium may therefore be due in part to inappropriate HKG selection

Full text not available from this repository.

More information

Accepted/In Press date: July 2012
e-pub ahead of print date: 13 July 2012
Published date: 13 July 2012
Keywords: Housekeeping genes, adhesions, mesothelium
Organisations: Human Development & Health

Identifiers

Local EPrints ID: 342835
URI: http://eprints.soton.ac.uk/id/eprint/342835
ISSN: 0015-0282
PURE UUID: 98b27893-a2f8-484a-9fc7-2b1eba6b785d
ORCID for Ying Cheong: ORCID iD orcid.org/0000-0001-7687-4597

Catalogue record

Date deposited: 17 Sep 2012 10:20
Last modified: 18 Jul 2017 05:26

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Contributors

Author: Khaled H. Sadek
Author: F.R. Cagampang
Author: K.D. Bruce
Author: N. Macklon
Author: Ying Cheong ORCID iD

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