蛋白酶体激活因子γ对未分化甲状腺癌细胞摄碘功能的影响

Influence of proteasome activator γ on iodine uptake capability of anaplastic thyroid carcinoma

目的 观察沉默蛋白酶体激活因子γ(REGγ)前后的甲状腺癌ARO及WRO细胞株的摄碘能力变化及摄碘相关特异蛋白mRNA水平变化,研究REGγ与甲状腺癌摄碘的关系。 方法 常规培养AROshN、AROshR、WROshN及WROshR甲状腺癌细胞株(shN为空白质粒,shR为沉默REGγ质粒),加入低剂量Na125I(3.7 kBq)后5、10、15、20、40和70 min测量细胞的摄碘率,观察细胞摄碘能力的变化,并利用实时荧光PCR检测摄碘相关特异蛋白[钠/碘转运体(NIS)、促甲状腺激素受体(TSHR)、甲状腺过氧化物酶(TPO)、甲状腺球蛋白(Tg)]mRNA水平的变化。采用配对t检验分析数据。 结果 沉默REGγ的AROshR及WROshR细胞摄碘能力均提高,其中AROshR细胞的提高率为128.1%[峰值(1 974±12)与(4 502±23)计数/min;t=17.30,P<0.05],WROshR细胞的提高率为67.4%[峰值(2 988±25)与(5 001±16)计数/min,t=13.20,P<0.05]。沉默REGγ的AROshR及WROshR细胞的NIS、TSHR、TPO、Tg mRNA表达均提高,其中在AROshR细胞中分别为AROshN细胞中的2.82、1.98、2.65和2.31倍,在WROshR细胞中分别为WROshN细胞中的2.21、1.78、2.51和1.78倍,与未沉默组相比差异均有统计学意义(t值:13.80~21.93,均P<0.05)。 结论 沉默REGγ可使甲状腺癌细胞摄碘相关特异蛋白上调表达,从而提高未分化型甲状腺癌细胞的摄碘能力。

Objective To investigate the changes of iodine uptake capability (IUC) and mRNA expression of iodine uptake-related proteins in ARO and WRO thyroid carcinoma cell lines after silence of proteasome activator γ (REGγ), and observe the relation between REGγ and IUC of thyroid carcinoma.Methods The AROshN, AROshR, WROshN and WROshR (shN= blank plasmid, shR=plasmid with silence of REGγ) thyroid carcinoma cell lines were routinely cultured. Low dosage (3.7 kBq) of Na125I was added and then IUC was determined at different time points (5, 10, 15, 20, 40 and 70 min). The mRNA expressions of sodium/iodine symporter (NIS), thyroid stimulating hormone receptor (TSHR), thyroid peroxidase (TPO) and thyroglobulin (Tg) were examined by real-time PCR. Paired t test was used to analyze the data.Results After the silence of REGγ, the peak values of IUC in AROshR and WROshR cells were increased from (1 974±12) to (4 502±23) counts/min, and from (2 988±25) to (5 001±16) counts/min, respectively. The increase rates were 128.1% in AROshR cells and 67.4% in WROshR cells (t values: 17.30, 13.20, both P<0.05). The mRNA expressions of NIS, TSHR, TPO, Tg in AROshR cells were 2.82, 1.98, 2.65 and 2.31 times higher than those in AROshN cells, and the expressions in WROshR cells were 2.21, 1.78, 2.51 and 1.78 times higher than those in WROshN cells (t values: 13.80-21.93, all P<0.05).Conclusion Silence of REGγ can increase the gene expressions of the iodine uptake-related proteins and elevate the IUC of thyroid carcinoma cells.