Journal of Endocrinological Investigation

, Volume 38, Issue 8, pp 875–884 | Cite as

Effects of monobutyl phthalate on steroidogenesis through steroidogenic acute regulatory protein regulated by transcription factors in mouse Leydig tumor cells

  • Y. Hu
  • C. Dong
  • M. Chen
  • Y. Chen
  • A. Gu
  • Y. Xia
  • H. Sun
  • Z. Li
  • Y. Wang
Original Article



Dibutyl phthalate (DBP) is one of the most widely used phthalate esters, and it is ubiquitous in the environment. DBP and its major metabolite, monobutyl phthalate (MBP), change steroid biosynthesis and impair male reproductive function. However, the regulatory mechanism underlying the steroid biosynthesis disruption by MBP is still unclear.


We analyzed the progesterone production, steroidogenic acute regulatory protein (StAR) mRNA, protein expression, and DNA-binding affinity of transcription factors (SF-1 and GATA-4).


Our results reveal that MBP inhibited progesterone production. At the same time, StAR mRNA and protein were decreased after MBP exposure. Furthermore, electrophoretic mobility shift assay showed that DNA-binding affinity of transcription factors (SF-1 and GATA-4) was decreased in a dose-dependent manner after MBP treatments. Western blot tests next confirmed that protein of SF-1 was decreased, but GATA-4 protein was unchanged. However, phosphorylated GATA-4 protein was decreased with 800 μM of MBP.


This study reveals an important and novel mechanism whereby SF-1 and GATA-4 may regulate StAR during MBP-induced steroidogenesis disruption.


Monobutyl phthalate Progesterone Steroidogenic acute regulatory protein Transcription factors Mouse Leydig tumor cells-1 



Bovine serum albumin


CCAAT/enhancer binding protein-β


Dosage-sensitive sex reversal adrenal hypoplasia critical region on chromosome X gene 1


Dibutyl phthalate




Fetal bovine serum


Monobutyl phthalate


Mouse Leydig tumor cells-1


3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide


Phthalate esters


Steroidogenic factor 1


Sodium dodecyl sulfate


Sterol regulatory element-binding protein


Serum-free medium


Steroidogenic acute regulatory protein



YH YW conceived and designed the experiments. CD MC YC performed the experiments. YH MC XH analyzed the data. AG YK HS contributed reagents/materials/analysis tools. YH wrote the paper. All authors read and approved the final manuscript. Research was funded by National Natural Science Foundation (81373041).

Conflict of interest

All the authors state that they have no conflicts of interest.

Ethical approval

Our research only used cell mode without Human Participants and/or Animals.

Informed consent

Not required.


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Copyright information

© Italian Society of Endocrinology (SIE) 2015

Authors and Affiliations

  • Y. Hu
    • 1
    • 2
  • C. Dong
    • 1
    • 2
  • M. Chen
    • 1
    • 2
  • Y. Chen
    • 3
  • A. Gu
    • 1
    • 2
  • Y. Xia
    • 1
    • 2
  • H. Sun
    • 1
    • 2
    • 4
  • Z. Li
    • 5
  • Y. Wang
    • 1
    • 6
  1. 1.State Key Laboratory of Reproductive Medicine, Institute of Toxicology, School of Public HealthNanjing Medical UniversityNanjingChina
  2. 2.Key Laboratory of Modern Toxicology of Ministry of Education, School of Public HealthNanjing Medical UniversityNanjingChina
  3. 3.Department of Molecular Cell Biology and Toxicology, Jiangsu Key Lab of Cancer Biomarkers, Prevention and Treatment, Cancer Center, School of Public HealthNanjing Medical UniversityNanjingChina
  4. 4.Jiangsu Provincial Center for Disease Control and PreventionNanjingChina
  5. 5.Department of Nutrition and Food Hygiene, School of Public HealthNanjing Medical UniversityNanjingChina
  6. 6.Safety Assessment and Research Center for Drug, Pesticide and Veterinary Drug of Jiangsu ProvinceNanjing Medical UniversityNanjingChina

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