Peptidylprolyl isomerase A (PPIA) as a preferred internal control over GAPDH and beta-actin in quantitative RNA analyses.

A good internal control is critical in all quantitative analyses of gene expression. Levels of bet-actin, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and peptidylprolyl isomerase A (PPIA) were analyzed in 78 samples (data obtained from our laboratory and from a publicly available database at http://www.ncbi.nlm.nih.gov/SAGE/). These libraries included cell lines and tissues from brain, breast, colon, kidney, ovary, pancreas, prostate, skin, and vascular origin. The level of PPIA mRNA is the most constant among the three genes. Hence, our study suggests that PPIA is a better internal control than beta-actin or GAPDH, the two most commonly used internal controls.

[1]  S. Hanash,et al.  Identification of androgen‐regulated genes in the prostate cancer cell line LNCaP by serial analysis of gene expression and proteomic analysis , 2001, Proteomics.

[2]  M. Schober,et al.  Identification of differentially expressed genes by serial analysis of gene expression in human prostate cancer. , 2001, Cancer research.

[3]  W. Lau,et al.  Differential gene expression of hepatocellular carcinoma using cDNA microarray analysis. , 2001, Oncology research.

[4]  Thomas D. Schmittgen,et al.  Effect of experimental treatment on housekeeping gene expression: validation by real-time, quantitative RT-PCR. , 2000, Journal of biochemical and biophysical methods.

[5]  E. Kroon,et al.  The housekeeping gene glyceraldehyde-3-phosphate dehydrogenase is inappropriate as internal control in comparative studies between skin tissue and cultured skin fibroblasts using Northern blot analysis , 1999, Archives of Dermatological Research.

[6]  D. Agapitos,et al.  Early changes of gene expression during cerulein supramaximal stimulation. , 1999, Pancreas.

[7]  J. Simons,et al.  Direct comparison of GAPDH, beta-actin, cyclophilin, and 28S rRNA as internal standards for quantifying RNA levels under hypoxia. , 1999, Biochemical and biophysical research communications.

[8]  K. Ranganna,et al.  Inhibition of platelet-derived growth factor BB-induced expression of glyceraldehyde-3-phosphate dehydrogenase by sodium butyrate in rat vascular smooth muscle cells. , 1997, Arteriosclerosis, thrombosis, and vascular biology.

[9]  S. Chouaib,et al.  Induction of tumor necrosis factor alpha expression in human T lymphocytes following ionizing gamma irradiation. , 1996, Journal of interferon & cytokine research : the official journal of the International Society for Interferon and Cytokine Research.

[10]  K. Kinzler,et al.  Serial Analysis of Gene Expression , 1995, Science.

[11]  W. A. Toscano,et al.  Transcriptional regulation of glyceraldehyde-3-phosphate dehydrogenase by 2,3,7,8-tetrachlorodibenzo-p-dioxin. , 1995, Biochemical and biophysical research communications.

[12]  N. Komori,et al.  Vitamin B6 deficiency causes activation of RNA polymerase and general enhancement of gene expression in rat liver , 1993, FEBS letters.

[13]  A. Partin,et al.  Association of glyceraldehyde-3-phosphate dehydrogenase expression with cell motility and metastatic potential of rat prostatic adenocarcinoma. , 1993, Cancer research.

[14]  N. Holbrook,et al.  Immediate and delayed molecular response of human keratinocytes to solar-simulated irradiation. , 1991, Laboratory investigation; a journal of technical methods and pathology.

[15]  M. Prystowsky,et al.  Glyceraldehyde-3-phosphate dehydrogenase mRNA is a major interleukin 2-induced transcript in a cloned T-helper lymphocyte. , 1990, Gene.

[16]  G. Woloschak,et al.  Modulation of gene expression in Syrian hamster embryo cells following ionizing radiation. , 1990, Cancer research.

[17]  A. Kimchi,et al.  Inhibitory effects of interferon on the expression of genes regulated by platelet-derived growth factor. , 1985, Proceedings of the National Academy of Sciences of the United States of America.

[18]  Eric Chicken,et al.  Nonparametric Statistical Methods: Hollander/Nonparametric Statistical Methods , 1973 .

[19]  E. Pitman A NOTE ON NORMAL CORRELATION , 1939 .

[20]  B. L. Welch THE SIGNIFICANCE OF THE DIFFERENCE BETWEEN TWO MEANS WHEN THE POPULATION VARIANCES ARE UNEQUAL , 1938 .

[21]  W. Taylor,et al.  Comparison of glyceraldehyde-3-phosphate dehydrogenase and 28S-ribosomal RNA gene expression as RNA loading controls for northern blot analysis of cell lines of varying malignant potential. , 1994, Analytical biochemistry.