Abstract
The COS-7 (CV-1 in Origin with SV40 genes) cells are known as non-steroidogenic cells because they are derived from kidney cells and the kidney is defined as a non-steroidogenic organ. Therefore, COS-7 cells are used for transfection experiments to analyze the actions of functional molecules including steroids. However, a preliminary study suggested that COS-7 cells metabolize [3H]testosterone to [3H]androstenedione. These results suggest that COS-7 cells are able to metabolize steroids. Therefore, the present study investigated the expression of steroidogenic enzymes and the metabolism of steroids in COS-7 cells. RT-PCR analyses demonstrated the expressions of several kinds of steroidogenic enzymes, such as cytochrome P450 side-chain cleavage enzyme, 3β-hydroxysteroid dehydrogenase/Δ5-Δ4 isomerase, cytochrome P450 7α-hydroxylase, cytochrome P450 17α-hydroxylase/17,20-lyase, 17β-hydroxysteroid dehydrogenase, 5α-reductase, cytochrome P450 21-hydroxylase, cytochrome P450 11β-hydroxylase, and cytochrome P450 aromatase in COS-7 cells. In addition, steroidogenic enzymes 3β-HSD, P4507α, 5α-reductase, P450c17, P450c21, P450c11β, and 17β-HSD actively metabolized various steroids in cultured COS-7 cells. Finally, we demonstrated that 17β-HSD activity toward androstenedione formation was greater than other steroidogenic enzyme activities. Our results provide new evidence that COS-7 cells express a series of steroidogenic enzyme mRNAs and actively metabolize a variety of steroids.
Highlights
The COS-7 (CV-1 in Origin with Simian Virus 40 (SV40) genes) cell line was developed by Prof
We further demonstrated that the mRNAs of several kinds of steroidogenic enzymes 3β-hydroxysteroid dehydrogenase/Δ5-Δ4 isomerase (3β-HSD; gene name Hsd3b), cytochrome P450 7α-hydroxylase (P4507α; gene name Cyp7b), cytochrome P450 17α-hydroxylase/17,20-lyase (P450c17; gene name Cyp17), 17β-hydroxysteroid dehydrogenase (17β-HSD; gene name Hsd17b), 5α-reductase, cytochrome P450 21-hydroxylase (P450c21; gene name Cyp21), cytochrome P450 11β-hydroxylase (P450c11β; gene name Cyp11b1), and cytochrome P450 aromatase (P450arom; gene name Cyp19) are expressed in COS-7 cells
Steroidogenic enzymes 3β-HSD, P4507α, 5α-reductase, P450c17, P450c21, P450c11β, and 17β-HSD were active in cultured COS-7 cells
Summary
It is derived from the CV-1 African green monkey kidney fibroblast cell line transformed by a mutant strain of Simian Virus 40 (SV40) that codes for the wild-type T-antigen[1,2]. This cell line has unique characteristics of fibroblast-like growth and virus susceptibility[1,2]. The expression of steroidogenic enzymes in the kidney of humans[19,20,21] and rodents[22,23,24] has been reported These results suggest that COS-7 cells may metabolize steroids. Primer StAR P450scc 3β-HSD type I 3β-HSD type VII P4507α P450c17 17β-HSD type I 17β-HSD type II 17β-HSD type III 17β-HSD type IV 5α-Reductase type I 5α-Reductase type II 5α-Reductase type III P450arom P450c11β P450c21
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