促甲状腺激素并非甲状腺乳头状癌发生发展的关键因素

Thyroid Stimulating Hormone may not be a Key Factor for the Development and Progression of Papillary Thyroid Cancer

目的:血清促甲状腺激素(TSH)在甲状腺乳头状癌(PTC)中的作用及机制尚不明确,本研究主要探讨TSH对甲状腺细胞系及乳头状癌细胞系的作用。方法:体外培养人甲状腺细胞系和乳头状癌细胞系,分别给予不同剂量(0 mU/L、5 mU/L及20 mU/L)的TSH干预。通过MTS及流式细胞术,观察TSH对甲状腺及乳头状癌细胞系增殖和细胞周期的作用;通过RNA-seq、ELISA检测TSH对细胞因子的影响;通过实时荧光定量PCR及Western blot寻找潜在的作用靶点。结果:MTS及流式细胞术结果显示,TPC-1和Nthy-ori-3-1细胞经TSH干预后增殖指数下降,20 mU/L浓度的TSH干预组细胞周期缩短。ELISA结果显示TPC-1中TSH下调CXCL8,上调CXCL10,而CXCL12的表达无明显变化。在Nthy-ori-3-1细胞中CXCL8和CXCL10的表达也观察到类似的结果,但CXCL12表达受到TSH的抑制。TSH可使Nthy-ori-3-1和Bcpap细胞中细胞命运决定因子(DACH1)的表达呈剂量依赖性上调,且TSH可抑制Bcpap中BRAF(V600E)以及Nthy-ori-3-1和TPC-1中BRAF的表达。结论:综上所述,我们并未发现TSH对甲状腺癌细胞有明显的促肿瘤作用。相反,本研究提示TSH可能呈部分抗癌作用。因此,TSH对甲状腺的致癌作用仍有待进一步研究。

Objective:The role and mechanism of serum thyroid-stimulating hormone(TSH)in thyroid papillary carcinoma(PTC)is still unclear.This study focused on the effect of TSH on thyroid cell lines and PTC cell lines.Methods:TSH was given to cultured human thyroid and PTC cell lines for various dosages(0 mU/L,5 mU/L and 20 mU/L).The effect of TSH on the proliferation and cell cycle of thyroid and papillary cancer cell lines was observed by MTS and flow cytometry.The effect of TSH on cytokines was detected by RNA-seq and ELISA.Potential targets were identified by real-time fluorescence quantitative PCR and Western blot.Results:MTS showed that the proliferation index decreased after TSH intervention in TPC-1 and Nthy-ori-3-1 cells.Besides,shortened cell cycle was only observed in the 20 mU/L TSH-intervention group.The results of ELISA indicated that TSH downregulated CXCL8 and upregulated CXCL10 in TPC-1,while no change was observed in CXCL12 expression.Similar results were also observed in CXCL8 and CXCL10 expression in Nthy-ori-3-1 cells.However,unlike the negative result of CXCL12 expression in TPC-1,its expression was inhibited by TSH in Nthy-ori-3-1 cells.Moreover,DACH1 expression was upregulated by TSH in a dose-dependent manner in Nthy-ori-3-1 and Bcpap cells,while BRAF(V600E)expression in Bcpap and BRAF expression in Nthy-ori-3-1 and TPC-1 were inhibited by TSH.Conclusion:In summary,we did not find obvious tumor promoting effect of TSH on thyroid cancer cell.Conversely,this study suggests that TSH may have a partial anticancer effect.Therefore,the carcinogenic effect of TSH on thyroid remains to be further studied.