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

The most suitable housekeeping genes within healthy and adhesion-mesothelial tissue are ACTB, YWHAZ, and CYC1, as they are most stable in both tissue types.

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Authors

Khaled Hassan Sadek, M.B.B.S., Felino Ramon Cagampang, Ph.D., Kimberley Davina Bruce, Ph.D., Nick Macklon, M.D., Ying Cheong, M.D.

Vol 98, Issue 4, Pages 1023-1027

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 PCR (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 stability may be tissue specific and affected by certain pathologies.

Design:

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

Setting:

Hospital.

Intervention (s):

Assessment of HKG expression stability using geNorm algorithm software.

Main Outcome Measures:

Stability measure (M) generated by geometric averaging of multiple target genes and mean pair-wise variation of genes.

Results:

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

Conclusion:

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 in inappropriate HKG selection.

Read the full text at: http://www.fertstert.org/article/S0015-0282(12)00686-3/fulltext


Fertility and Sterility

Editorial Office, American Society for Reproductive Medicine

Fertility and Sterility® is an international journal for obstetricians, gynecologists, reproductive endocrinologists, urologists, basic scientists and others who treat and investigate problems of infertility and human reproductive disorders. The journal publishes juried original scientific articles in clinical and laboratory research relevant to reproductive endocrinology, urology, andrology, physiology, immunology, genetics, contraception, and menopause. Fertility and Sterility® encourages and supports meaningful basic and clinical research, and facilitates and promotes excellence in professional education, in the field of reproductive medicine.

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