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Molecular Biology Reports

, Volume 39, Issue 12, pp 10019–10030 | Cite as

A newly identified TSHβ splice variant is involved in the pathology of Hashimoto’s thyroiditis

  • ChunRong Liu
  • LanYing Li
  • Fan Ying
  • CangDan Xu
  • XiaoYi Zang
  • ZhiHong Gao
Article

Abstract

Thyrotropin (TSH) is a protein that plays a key role in the control of thyroid function. TSH consists of a common α-subunit and a unique β-subunit; the latter is responsible for hormone specificity. A novel splice variant of human TSHβ was identified in 2009. To date, only the tissue distribution of the human TSHβ splice variant mRNA has been studied. Therefore, we aimed to characterize the protein translated from this splice variant. Salting-out, dialysis and concentration of serum proteins were followed by immunoprecipitation to identify the hTSHβ splice variant in serum. Stable CHO cell lines expressing the hTSHβ splice variant and V5-hTSHα were generated. After co-culture, co-immunoprecipitation was used to determine if the hTSHβ splice variant can dimerise with TSHα. We showed for the first time that the hTSHβ splice variant exists in human serum and dimerises with TSHα. To explore the relationship between the TSHβ splice variant and the pathogenesis of autoimmune thyroiditis, we assessed variations in the mRNA expression of the TSHβ splice variant in the peripheral blood leukocytes (PBLs) of Hashimoto’s thyroiditis (HT) patients using quantitative RT-PCR. We found that the mRNA expression levels of the TSHβ splice variant were higher in the PBLs of HT patients who were not undergoing prednisone therapy (n = 10, P < 0.0001) and in the PBLs of HT patients with a longer duration of illness (>18 months) who were undergoing prednisone therapy (n = 5, P = 0.023) than in those of the control group. This pattern was reversed in the PBLs of HT patients with a shorter duration of illness (<9 months) who were undergoing prednisone therapy (n = 8, P < 0.0001). Dexamethasone inhibition of the TSHβ splice variant mRNA expression occurred in a dose- and time-dependent manner. These results demonstrated that the TSHβ splice variant may participate in the pathogenesis of HT.

Keywords

Dimer Human Serum TSHα TSHβ splice variant Hashimoto’s thyroiditis 

Abbreviations

TSH

Thyrotropin

BCA

Bicinchoninic acid assay

CHO

Chinese hamster ovary cell

IP

Immunoprecipitation

RT-PCR

Reverse transcription-polymerase chain reaction

TCA

Trichloroacetic acid

PBL

Peripheral blood leukocyte

HT

Hashimoto’s thyroiditis

Notes

Acknowledgments

We are very grateful to Dr. Dong-chun Liang for technical assistance and Dr. Bao-li Wang for English revision. This research was supported by Grants (30972559) from the National Natural Science Foundation of China.

Conflict of interest

The authors declare that there is no conflict of interest that would prejudice the impartiality of this scientific work.

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Copyright information

© Springer Science+Business Media B.V. 2012

Authors and Affiliations

  • ChunRong Liu
    • 1
  • LanYing Li
    • 1
  • Fan Ying
    • 1
  • CangDan Xu
    • 1
  • XiaoYi Zang
    • 1
  • ZhiHong Gao
    • 1
  1. 1.Key Laboratory of Hormones and Development (Ministry of Health)Metabolic Diseases Hospital & Tianjin Institute of Endocrinology, Tianjin Medical UniversityTianjinChina

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