FRTL-5 Rat Thyroid Cells Release Thyroglobulin Sequestered in Exosomes: A Possible Novel Mechanism for Thyroglobulin Processing in the Thyroid.

IF 1.7 Q4 ENDOCRINOLOGY & METABOLISM Journal of Thyroid Research Pub Date : 2016-01-01 Epub Date: 2016-06-09 DOI:10.1155/2016/9276402

Pavel Vlasov, Sonia Q Doi, Donald F Sellitti

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引用次数: 8

Abstract

Exosomes are 30-100 nm, membrane-bound vesicles containing specific cellular proteins, mRNAs, and microRNAs that take part in intercellular communication between cells. A possible role for exosomes in thyroid function has not been fully explored. In the present study, FRTL-5 rat thyroid cells were grown to confluence and received medium containing either thyroid stimulating hormone (TSH), exogenous bovine thyroglobulin (bTg), or neither additive for 24 or 48 hours followed by collection of spent medium and ultracentrifugation to isolate small vesicles. Transmission electron microscopy and Western blotting for CD9 indicated the presence of exosomes. Western blotting of exosome extract using a monoclonal anti-Tg antibody revealed a Tg-positive band at ~330 kDa (the expected size of monomeric Tg) with a higher density in TSH-treated cells compared to that in untreated cells. These results are the first to show that normal thyroid cells in culture produce exosomes containing undegraded Tg.

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FRTL-5大鼠甲状腺细胞释放外泌体中隔离的甲状腺球蛋白:甲状腺中甲状腺球蛋白加工的可能新机制

外泌体是30-100 nm的膜结合囊泡，含有特定的细胞蛋白、mrna和microrna，参与细胞间的细胞间通讯。外泌体在甲状腺功能中的可能作用尚未得到充分探讨。本研究将FRTL-5大鼠甲状腺细胞培养至合流状态，并接受含有促甲状腺激素(TSH)、外源性牛甲状腺球蛋白(bTg)或不含任何添加剂的培养基，培养24或48小时，然后收集培养基并进行超离心分离小泡。CD9的透射电镜和Western blotting显示外泌体的存在。使用单克隆抗Tg抗体对外泌体提取物进行Western blotting，发现tsh处理的细胞中有一个约330 kDa(单体Tg的预期大小)的Tg阳性带，与未处理的细胞相比，tsh处理的细胞密度更高。这些结果首次表明，正常的甲状腺细胞在培养中产生含有未降解Tg的外泌体。

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