Expression of Three Negative Regulators ofCYP17Gene Transcription in Adrenocortical Cells

Abstract

The expression of negative regulators of CYP17 gene expression: DAX-1, COUP-TF and N-CoR was investigated in bovine adrenocortical cells in primary culture. The cells were incubated for 6 hours in a defined medium, containing ACTH (10 nM) or forskolin (25 microM). Total RNA was isolated, the concentration of CYP17 gene transcript was determined by hybridization analysis, and polyA RNA was reverse-transcribed to obtain cDNA. Fragments of CYP17, DAX-1, COUP-TF, N-CoR and beta-actin cDNA were amplified by PCR and the amplified cDNAs were analyzed by agarose gel electrophoresis followed by densitometry. It was found that DAX-1 and COUP-TF mRNA levels were unaffected by treatment with ACTH or forskolin. On the contrary, both compounds enhanced the level of mRNA encoding N-CoR. Under the same conditions, ACTH or forskolin substantially stimulated CYP 17 mRNA accumulation. Our results suggest that in the adrenocortical cells in culture, the expression of DAX1 and COUP-TF is constitutive, whereas N-CoR expression might be inducible.

The Effects of Functional Polymorphisms in the TGFβ1 Gene on Nasopharyngeal Carcinoma Susceptibility

“…All analyses were done in a blinded fashion with the laboratory persons unaware of genotyping data. The expression of individual TGFβ1 measurements was calculated relative to expression of β ‐actin using a modification of the method described by Lehmann et al 15 …”

“…All analyses were done in a blinded fashion with the laboratory persons unaware of genotyping data. The expression of individual TGFβ1 measurements was calculated relative to expression of β-actin using a modification of the method described by Lehmann et al 15 Statistical Analysis χ 2 tests were used to examine the differences in the distributions of genotypes between cases and controls. The association between the TGFβ1 -509C/T and 869T/C polymorphisms and risk of NPC was estimated by odds ratios (ORs) and their 95% confidence intervals (CIs), which were calculated by unconditional logistic regression models adjusted for age and sex.…”