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Unité Mixte de Recherche 6547 CNRS-Université Blaise Pascal Clermont II (A.-M.L.-M., P.V., C.T., G.V., C.J., A.M.), Génétique des Eucaryotes et Endocrinologie Moléculaire, Complexe Universitaire des Cézeaux, 63177 Aubière, France; Institut National de la Santé et de la Recherche Médicale U-576, Département d’Endocrinologie (J.B., X.B.), Institut Cochin, Université René Descartes-Paris V, 75014 Paris, France; Department of Endocrinology (N.G.-P.), Faculty of Medicine, University of Sherbrooke, Québec J1H 5N4, Canada; and Protein Function Discovery Facility (D.H.), Queen’s University, Ontario K7L 3N6, Kingston, Canada
Address all correspondence and requests for reprints to: Dr. A. Martinez, UMR6547 CNRS-Université Blaise Pascal Clermont II, Génétique des Eucaryotes et Endocrinologie Moléculaire, Complexe Universitaire des Cézeaux, 24 avenue des Landais, 63177 Aubière Cedex, France. E-mail: antoine.martinez{at}univ-bpclermont.fr.
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Abstract |
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Introduction |
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The aldo-keto reductases (AKR) belong to an oxidoreductase superfamily that catalyze the reduction of a wide variety of substrates including aldoses, aliphatic and aromatic aldehydes and ketones, prostaglandins, and xenobiotics (14). Among the AKR1B subfamily, aldose reductase (AKR1B1 in human, AKR1B3 in mouse), known as the first enzyme of the polyol pathway of sugar metabolism, is one of the most studied because of its role in diabetic complications (15, 16). Although being ubiquitously expressed in both species, the most abundant source of aldose reductase in human tissues is the adrenal gland (17). This suggests an important role for AKR1B1 in this specialized organ in which its isocaproaldehyde reductase activity could be recruited (18). In human and rodent adrenals, the toxicity of isocaproaldehyde, the product of cholesterol side chain cleavage by the cytochrome P450scc (CYP11A gene), is mainly neutralized by the reductase activity of AKR1B1 and AKR1B7/mouse vas deferens protein (MVDP), respectively (18, 19). Moreover, we have shown that in rodents, the akr1b7 gene expression is under control of ACTH through cAMP pathway (20, 21, 22, 23). Thus, full steroidogenic activity should require the coordinate regulation by ACTH/cAMP of genes involved in cholesterol transport and steroid conversion but also of scavenger genes detoxifying harmful byproducts of steroidogenesis e.g. isocaproaldehyde (19) and free radicals (24). Until now the murine AKR1B7 is the only aldose reductase-like protein showing ACTH responsiveness. A possible regulation of the aldose reductase AKR1B1 by ACTH/cAMP has not been investigated yet in human adrenal. To shed more light on the role of AKR1B1 in adrenal steroidogenesis and pathophysiology, we compared the expression of AKR1B1 gene with those of genes involved in steroidogenesis e.g. steroidogenic acute regulatory protein gene (StAR) and CYP11A (25, 26, 27), or in adrenal differentiation, e.g. steroidogenic factor (SF)-1 and dosage-sensitive sex reversal-adrenal hypoplasia congenita critical region on the X chromosome, gene 1 (DAX1) (28, 29), in human adrenocortical tumor cell line and malignant or benign adrenocortical neoplasms with different endocrine profiles. These comparisons should reveal whether AKR1B1 along with steroidogenic genes is coordinately controlled and whether its expression could be correlated with the malignancy and/or the endocrine status of the tumors.
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Materials and Methods |
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Adrenal tissues were obtained during surgery and were immediately dissected by the pathologist, frozen, and stored in liquid nitrogen until use. Hormonal investigation and diagnosis were performed as previously reported (30). For malignant tumors staging was performed as previously reported (31). Adrenal tumors were diagnosed by the use of classical histological criteria and molecular genetic markers as previously reported by the Cortico et Medullosurrénale: Etude des Tumeurs Endocrines (COMETE) network (13), which is dedicated to the study of adrenal tumors. All the carcinomas are primitive tumors and exhibited IGF-II overexpression and a histological Weiss score 4 or more. None of the patients with carcinomas have been submitted to mitotane therapy before surgery. Among these patients during a 6- to 24-month follow-up period, five (numbered b, 2, 37, 39, and 40 in Table 1
) presented with tumor recurrence or distant metastasis. The Weiss score was 1 or less in all adrenal adenomas. Normal adrenal cortex tissue was obtained from normal glands removed during the surgery of adjacent nonendocrine tumors (kidney tumors or incidentalomas). Aldosterone adenomas were removed from patients exhibiting a primary hyperaldosteronism and a small benign adrenocortical adenoma (all these Conn’s adenomas have a diameter 
15 mm). Adrenal tissues from ACTH-dependent Cushing’s syndrome were obtained during bilateral adrenalectomy performed for Cushing disease. Informed consent was given for adrenal tissue collection as part of a protocol approved by the Institutional Review Board of the Cochin Hospital.
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Recombinant protein production
AKR1B1 cDNA was isolated by RT-PCR, starting from 2 µg human adrenal total RNAs, using Moloney murine leukemia virus reverse transcriptase (Promega, Charbonnier, France) and Taq polymerase (Promega) and outside primers containing engineered 5' B1 NdeI primer (5'-CGGCAGCCATATGGCAAGCCGTC-3') and 3' B1 EcoRI primer (5'-CGGAATTCGGGCTTCAAAACTCTTCATGG-3'). AKR1B7 cDNA was obtained by PCR amplification with MVDP pUC13 (33) as template and with outside primers containing engineered 5' B7 NdeI primer (5'-CGGCAGCCATATGGCCACCTTCGT-3') and 3' B7 BamHI primer (5'-CGGCATCCCGTCAGTATTCCTCGTGG-3').
AKR1B8 cDNA was isolated by RT-PCR, starting from 2 µg mouse adrenal total RNAs, using Moloney murine leukemia virus reverse transcriptase and Taq polymerase (Promega) and outside primers containing engineered 5' B8 NdeI primer (5'-CGGCAGCCATATGGCCACGTTCGTGG-3') and 3' B8 EcoRI primer (5'-CGGGATCCCGGGGCTGACTCAGCTTCA-3').
Recombinant AKR1B1, AKR1B7, and AKR1B8 were expressed in Escherichia coli after inserting their corresponding cDNA into the Nde1 and EcoRI sites of the PET 28a vector (Novagen, Tebu, Le Perray-en-Yvelines, France) to produce N-terminal fusions with six histidine residues. AKR1B10 cDNA was inserted into PET 16 vector (gift of Dr. D Hyndman, Queen’s University, Ontario, Canada). Recombinant AKR1B1, AKR1B7, AKR1B8, and AKR1B10 proteins were produced in BL21(DE3) pLys S cells upon isopropyl-ß-D-thiogalactopyranoside induction and purified by nickel affinity chromatography according to the manufacturer’s instructions (Novagen). For each protein, column fractions were analyzed by SDS-PAGE, and those containing the purified protein were pooled and stored at 4 C.
Antibodies and Western blot experiments
For production of the L3 antibody, rabbits were injected with a glutathione S-transferase fusion of the 17 C-terminal amino acid residues of the murine AKR1B7 protein, and the antibody was obtained and tested as previously described (34). L3 antibody is both specific for murine AKR1B7 and human AKR1B1 (Fig. 1). Western blots were performed as previously described (22). Blots were treated with primary L3 antibody at a 1:3000 dilution for 1 h at room temperature. Peroxidase-conjugated antimouse or antirabbit secondary antibodies were added at 1:15000 dilution for 1 h at room temperature. Peroxidase activity was detected with the enhanced chemiluminescence system (Amersham Biosiences, Buckinghamshire, UK). Densitometric analysis of the immunoreactive protein bands obtained in Western blots were performed on nonsaturated signals using Molecular Analyst software (Bio-Rad Laboratories, Inc., Marnes la Coquette, France).
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Human NCI-H295 (35) adrenocortical cells were routinely cultured in DMEM/nutrient mixture F12 (DMEM-F12) supplemented with 3% fetal calf serum, 2% Ultroser G (InVitrogen, Cergy-Pontoise, France), 1% ITS (insulin, transferrin, sodium selenite) (InVitrogen), penicillin (100 U/ml), and streptomycin (100 µg/ml). Human placental JEG-3 cells were grown in DMEM-F12 supplemented with 10% fetal calf serum, penicillin (100 U/ml), and streptomycin (100 µg/ml). When required, 10–5 M forskolin were added in the culture medium for the indicated time.
Northern blot analysis
Total RNAs from human biopsies or cellular samples were individually extracted with trizol (InVitrogen). Northern blots were individually performed with 25 µg of the RNAs as described previously (22). Probes used were the 3' untranslated region of AKR1B1 cDNA segment from position 1032 to 1350 pb, the human StAR cDNA fragment from position 467 to 908, and the bovine CYP11A pvuII cDNA fragment of 1.8 kb (kindly provided by Dr. Waterman M, Vanderbilt University, Nashville, TN). To normalize the loading of RNAs, Northern blots were stripped and rehybridized with a mouse ß-actin probe extracted from pGEM-7ZF-ß-actin by EcoRI/BamHI digestion. Hybridization signals were quantified by phosphor imager using Quantity One software (Bio-Rad Laboratories).
Enzymatic assays
The standard reaction mixture for the reductase activities contained 0.1 M sodium phosphate buffer (pH 6.6), 0.4 M ammonium sulfate, 100 mM nicotinamide adenine dinucleotide phosphate reduced (NADPH2), appropriate isocaproaldehyde, and 5 µg of recombinant AKR1B1 or 200 µg of NCI-H295 cytosolic proteins. The reaction was carried out at 25 C, and the decrease in NADPH2 was monitored by spectrophotometer at 340 nm. Reactions were routinely started by the addition of enzyme or protein extracts. Controls without substrate or without enzyme were run simultaneously. One enzyme unit is defined as the change at 340 nm corresponding to the oxidation of 1 µmol NADPH2.
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Results |
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The ACTH-responsive-AKR1B7 protein is responsible for the reduction of isocaproaldehyde in mouse adrenocortical cells (19). Such an isocaproaldehyde reductase function has not been assigned in the human adrenal cortex yet, although AKR1B1, the human aldose reductase, is able to reduce isocaproaldehyde in vitro (18). We thus undertook the isolation of a human ortholog for the akr1b7 gene. Attempts to isolate AKR1B7 cDNA in human adrenal cortex samples by repeated RT-PCR experiments using primers complementary to conserved sequences within AKR1B subfamily or by bioinformatic searches within genomic or EST databases using BLAST failed to isolate a strict ortholog (not shown). Indeed, extensive sequence analyses of RT-PCR products showed that all amplified cDNAs corresponded to AKR1B1.
It has been previously shown that the C-terminal region of aldose reductase is critical to substrate specificity (36). In agreement with these data, the C-terminal domains were divergent among the AKR1B subfamily members (Fig. 1A
). Interestingly, in the C-terminal region, AKR1B7 exhibited more homologies with human aldose reductase AKR1B1 (82%) than with mouse aldose reductase AKR1B3 (65%), suggesting that AKR1B7 and AKR1B1 might have similar substrate specificities.
Polyclonal antibodies were raised against AKR1B7 C-terminal peptide. As illustrated by the Western blot analysis shown in Fig. 1B, both AKR1B7 and AKR1B1 recombinant proteins were strongly recognized by the anti-AKR1B7 antibody (Fig. 1B). The antibody does not cross-react with AKR1B3 or AKR1B8 and cross-reacts only slightly with AKR1B10. Because akr1b7 gene expression, first detected at embryonic d 13.5, was shown to follow the onset of glucocorticoid synthesis in mouse fetal adrenal (37), we looked at AKR1B1 expression in developing adrenals of human fetuses. The Western blot in Fig. 1C showed that AKR1B1 is detected in adrenals from fetuses aged 15–19 wk, a time at which fetal adrenal is potentially capable to produce cortisol (38).
Altogether these data prompted us to conclude that there was no akr1b7/mvdp ortholog gene in human, and we postulated that AKR1B1 might fulfill the same role in human adrenals.
AKR1B1 expression in human adrenocortical cells is regulated by cAMP
To examine a possible regulation of AKR1B1 expression by the protein kinase A (PKA) transduction pathway, steady-state levels of AKR1B1 mRNA were measured in human NC1-H295 cells cultured for increasing periods of time in the presence of forskolin, an activator of cAMP synthesis, that was preferred to the natural inducer because these cells are poorly responsive to ACTH (39). The pattern of forskolin induction was compared with that of steroidogenic genes such as CYP11A and StAR. The action of forskolin was time dependent and first detectable after 6 h of treatment (Fig. 2, A and C). Maximal induction of AKR1B1 mRNA was 2.1-fold (P < 0.05, compared with untreated cells) after 12 h. The kinetic and magnitude of forskolin induction of CYP11A mRNA level followed a very similar pattern. Note that StAR mRNA level showed only a 1.3-fold increase on forskolin induction. The low responsiveness of StAR in NCI-H295 cells was already mentioned (40). AKR1B1 cDNA had first been cloned in the human placenta (41). As expected, AKR1B1 was expressed in trophoblastic JEG-3 cells (Fig. 2B). Interestingly, forskolin did not affect AKR1B1 mRNA expression in these cells, indicating that the forskolin responsiveness seems to be at least specific of the adrenocortical cells.
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Aldose reductase (AKR1B1) has been described as a major reductase for isocaproaldehyde in adrenal glands (19). As shown in Fig. 2, forskolin (10–5 M) induced a significant increase in AKR1B1 mRNA levels. To assess the physiological relevance of this effect, isocaproaldehyde reductase activity was measured in NCI-H295 adrenocortical cells. Figure 3A shows that the recombinant AKR1B1 had the ability to reduce isocaproaldehyde with a constant of molecular activity (Kcat) value in accordance with those previously reported (18). Recombinant AKR1B1 isocaproaldehyde reductase activity was strongly inhibited by sorbinil, a potent inhibitor of aldose reductase (Fig. 3A). In contrast, AKR1B7 showed no sensitivity to this compound (19). Isocaproaldehyde reductase activities, measured in cytosolic protein extracts from NCI-H295 cells, were significantly enhanced by forskolin treatment and inhibited by sorbinil (Fig. 3B).
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Table 1 outlined clinical data from the patients examined. Expression of SF-1, DAX1, CYP11A, StAR, and AKR1B1 mRNAs was analyzed by Northern blot experiments in 20 of the 37 tissues available for RNA extraction. Although the levels of their mRNAs varied among the tumors, expression of SF-1, DAX1, CYP11A, StAR, and AKR1B1 genes was detectable in all samples analyzed (Fig. 4). The relative amounts of mRNAs from all specimens studied were measured (Table 2) and did not correlate with endocrine activity (Tables 1 and 2). When considered together, data from cortisol-producing adenomas (CPAs) and nonfunctional adenomas (NFAs) revealed that the relative levels of all the tested genes, with the exception of DAX1, were significantly higher in this group than in malignant tumors [adrenocortical carcinomas (ACCs)] (Fig. 5). In accordance with previous data (42), relative mRNA levels of DAX1 were significantly lower in aldosterone-producing adenomas (APAs) than in other adenomas (CPA+NFA).
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). In contrast, ACCs had low to barely undetectable levels of AKR1B1 mRNA, compared with all other groups (Fig. 4). If individual values from the population of tumors were divided into benign or malignant, statistical analysis revealed that the relative AKR1B1 mRNA levels measured in carcinomas (ACCs) (0.95 ± 0.2 SEM) were significantly lower (P < 0.001) than the overall mean value calculated from adrenocortical adenomas (CPA+NFA+APA) (9.29 ± 3.05). Moreover, as shown in Fig. 5, the values measured in carcinomas were lower than those determined in CPAs plus nonfunctional adenomas (P < 0.01) as well as in APAs (P < 0.05). The relative values measured in ACCs slightly varied, from 0.5 to 1.3 arbitrary units, and were always lower than those observed in other pathological adrenal tissues (from 2.2 to 21.0). No significant differences in relative AKR1B1 mRNA levels were measured in CPAs (15.1 ± 2.9) and nonfunctional adenomas (9.5 ± 2.5), suggesting that there was no correlation between expression of the AKR1B1 gene and the hormonal status of the neoplasm. AKR1B1 protein content is decreased in ACCs
AKR1B1 protein expression was evaluated by Western blotting using the antibodies described above. As shown in Fig. 6A, AKR1B1 could be detected in both normal and most of neoplastic adrenal tissues but with marked quantitative differences. AKR1B1 was markedly accumulated in CPAs, NFAs, APAs, and Cushing’s hyperplasias. By contrast, most ACCs (seven of eight) exhibited low or hardly detectable amounts of AKR1B1 protein. Quantification of AKR1B1 signals showed significantly lower concentrations of the enzyme in malignant tumors than in normal tissue, adenomas, or Cushing’s hyperplasias (Fig. 6B). The results of Western blot studies were consistent with those of mRNA assays.
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Discussion |
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In accordance with a previous report (40), our data showed that expression of CYP11A and StAR genes, both involved in the rate-limiting step of steroidogenesis, did not correlate with the endocrine profiles of patients examined. In the same way, we confirmed that SF-1 mRNA levels did not reflect the hormonal status of the neoplasms (45, 46) and that low DAX1 expression seemed to be associated with mineralocorticoid-producing adenomas (42). The implication of SF-1 in the regulation of DAX1 has been first reported in adrenal cell transfection studies (47) and then formally established in vivo in the developing gonad (48). We did not observe such a positive link between the two orphan receptors expression in the adrenocortical tumors. Other mechanisms might probably participate to DAX1 regulation in neoplastic tissues or as suggested by Hoyle et al. (48), the SF-1 requirement for DAX1 expression could vary during development, being essential during the embryonic period but not in the adult organ. Low expression of transcriptional repressors acting on SF-1 target genes seems to be a more general feature of APAs because low chicken ovalbumin upstream promoter transcription factor I expression was also found in these tumors (46). However, mechanisms independent of SF-1 might also occur because CYP11B2 was shown to be not positively regulated by SF-1 (49) [nor was overexpressed in APA (50)]. By contrast, cortisol hypersecretion in adenomas could result from disordered expression of SF-1 target genes encoding proteins acting at downstream steps, e.g. CYP17, rather than at initial steps of steroidogenesis (StAR or CYP11A). Indeed, excessive cortisol production in CPAs was shown to correlate with the overexpression of CYP17 in a manner reciprocal to that of its repressors DAX1 and COUP-TFs (51).
The most prominent finding of this study is that AKR1B1 gene is differentially expressed in benign vs. malignant tumors in adrenal cortex. AKR1B1 gene expression, measured at mRNA and protein levels, is strongly decreased in ACCs, compared with that in adenomas. Two reasons may account for low AKR1B1 expression in carcinomas. First, chromosomal alterations, including mutations or rearrangements, could reduce expression of the gene. However, no chromosomal abnormalities concerning the 7q35 region (the aldose reductase gene locus) (52) had ever been described in adrenal tumorigenesis (7). Alternatively, inhibition of AKR1B1 gene expression, in carcinomas, may be due to dysregulation of the mechanisms underlying the control gene expression. The mechanisms regulating expression of AKR1B1 in adrenals were unknown until the present report. Indeed, data reported herein suggest that cAMP is a regulator of AKR1B1 expression in human adrenocortical cells. Interestingly, the transcription factor cAMP-responsive element-binding protein (CREB) was shown to be strongly decreased at the protein level in ACCs (53). A loss of expression of the ACTH receptor have been reported in adrenal cancer (54), and cAMP stimulated PKA activity is lower in adrenal cancers than adenomas (55). This could take part in the decreased expression of AKR1B1. However, the factors that induced this inhibition of AKR1B1 in ACCs remain to be more thoroughly determined.
Changes in AKR1B1 mRNA level in response to cAMP, in parallel with steroidogenic genes, suggest that AKR1B1 may be considered a marker of adrenocortical cell differentiation. There are some evidences that members of the AKR superfamily may be associated with cancer progression. A previous report (56) has shown that an aldose reductase-like protein was induced during rat hepatocarcinogenesis. Additionally, AKR1B1 and AKR1B10 are overexpressed by 29 and 54%, respectively, in some human liver cancers (57). The physiological function of aldose reductase is still unclear, and recent data have suggested that its main role may be detoxication of reactive aldehydes. Oxidative stress plays an important role in various pathological states including cancer (58). HNE is believed to be responsible for the cellular pathological effects observed during oxidative stress in vivo (59), and HNE protein adducts have been detected in human renal cell carcinomas (60). HNE exhibited a wide range of biological activities, including stimulation of phospholipase C, stimulation of neutrophil migration, and reduction of gap-junction communication (61, 62). Interestingly, it has been reported that 
1-connexin 43 gap junctions were decreased in the human malignant adrenocortical tumors (63). AKR1B1 catalyzes the reduction of HNE, suggesting that this enzyme may be a part of the cellular defenses against oxidative stress in physiological and pathological conditions (19, 62). On the contrary, overexpression of aldose reductase and/or aldose reductase-like proteins in hepatomas has been interpreted as a defense reaction against harmful metabolites produced by the growing cancer cells (56). Whatever the mechanisms that lead to down-regulation of AKR1B1 in adrenocortical carcinomas, further studies are needed to elucidate whether the decrease in AKR1B1 expression in malignant tumors is merely a consequence of a general dedifferentiation of the tumor or whether it contributes to the pathogenesis of this disease by promoting, for instance, as a consequence of HNE accumulation, alterations in cell-cell communication through progressive loss of gap junctions.
There are no reliable criteria for accurately distinguishing between benign and malignant adrenocortical tumors, although certain markers have been proposed to be specific of adrenocortical carcinomas. For example, it has been reported that malignant adrenocortical tumors expressed low levels of StAR mRNA, compared with adenomas (40). Other reports, however, showed that StAR expression was about equal in adrenocortical adenomas and carcinomas (42, 64). Immunohistochemical studies have shown that GATA-4 (a zinc finger transcription factor) and Ki-67 (a cell cycle-associated marker) were overexpressed in adrenal carcinomas (65, 66). The angiogenic factor, vascular endothelial growth factor, and the protein thrombospondin-1 may represent possible markers of the transition of ACCs toward malignancy. Vascular endothelial growth factor concentrations were increased in carcinomas, whereas those of thrombospondin-1 were decreased (67). Overexpression of IGF-II, IGF-binding protein-2, and IGF-I receptor has been associated with malignancy (68, 69). However, there is no perfect marker, and the combined use of multiple indicators of malignancy is required to advance our understanding of both normal and pathological adrenocortical physiology. Although the usefulness of AKR1B1 has to be validated on a larger cohort and compared with standard criteria of histological diagnosis, several evidences point out that it could be chosen as a good candidate marker for further studies. Firstly, 88% of the carcinomas exhibit AKR1B1 protein concentrations below the lowest value measured in normal tissues and benign neoplasms. Second, AKR1B1 is a very stable soluble protein whose detection is easy to carry out simply using a Western blot analysis, although more sensitive quantitative analysis could be easily improved by development of an AKR1B1-based RIA or ELISA. Finally, induction of AKR1B1 mRNA levels by forskolin suggests that its expression is under ACTH control. Because AKR1B1 is a major reductase for reactive aldehydes formed during steroidogenesis and lipid peroxidation (19, 62), it will be worth investigating whether maintaining a high capacity of AKR1B1-dependent detoxication could impair or delay malignant transformation process in adrenal cortex. It would be interesting to further evaluate the value of AKR1B1 for differential diagnosis between adenomas and carcinomas and its prognosis value on a larger cohort and during prospective studies.
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Footnotes |
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Abbreviations: ACC, Adrenocortical carcinoma; APA, aldosterone-producing adenoma; CPA, cortisol-producing adenoma; DAX1, dosage-sensitive sex reversal-adrenal hypoplasia congenita critical region on the X chromosome, gene 1; HNE, 4-hydroxynonenal; MVDP, mouse vas deferens protein; NADPH, nicotinamide adenine dinucleotide phosphate reduced; NFA, nonfunctional adenoma; PKA, protein kinase A; SF, steroidogenic factor; StAR, steroidogenic acute regulatory protein.
Received October 21, 2003.
Accepted March 10, 2004.
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1 Connexin 43 gap junctions are decreased in human adrenocortical tumors. J Clin Endocrinol Metab 85:890–895This article has been cited by other articles:
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  M.-C. Battista, C. Roberge, A. Martinez, and N. Gallo-Payet 24-Dehydrocholesterol Reductase/Seladin-1: A Key Protein Differentially Involved in Adrenocorticotropin Effects Observed in Human and Rat Adrenal Cortex Endocrinology, September 1, 2009; 150(9): 4180 - 4190. [Abstract] [Full Text] [PDF]
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 Z. Kabututu, M. Manin, J.-C. Pointud, T. Maruyama, N. Nagata, S. Lambert, A.-M. Lefrancois-Martinez, A. Martinez, and Y. Urade Prostaglandin F2{alpha} Synthase Activities of Aldo-Keto Reductase 1B1, 1B3 and 1B7 J. Biochem., February 1, 2009; 145(2): 161 - 168. [Abstract] [Full Text] [PDF]
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B. Ragazzon, A.-M. Lefrancois-Martinez, P. Val, I. Sahut-Barnola, C. Tournaire, C. Chambon, J.-L. Gachancard-Bouya, R.-J. Begue, G. Veyssiere, and A. Martinez Adrenocorticotropin-Dependent Changes in SF-1/DAX-1 Ratio Influence Steroidogenic Genes Expression in a Novel Model of Glucocorticoid-Producing Adrenocortical Cell Lines Derived from Targeted Tumorigenesis Endocrinology, April 1, 2006; 147(4): 1805 - 1818. [Abstract] [Full Text] [PDF]
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S. R. Bornstein and P. J. Hornsby What Can We Learn from Gene Expression Profiling for Adrenal Tumor Management? J. Clin. Endocrinol. Metab., March 1, 2005; 90(3): 1900 - 1902. [Full Text] [PDF]
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