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    高碘通过调控钠碘转运体基因启动子甲基化水平对甲状腺乳头状癌细胞恶性生物学行为的影响

    Effect of high iodine on the malignant biological behavior of papillary thyroid carcinoma cells by regulating the methylation level of sodium iodine transporter gene promoter

      摘要
    • 摘要:
      目的 通过体外细胞实验探究高碘对甲状腺乳头状癌(PTC)发展的影响，及钠碘转运体(NIS)基因启动子甲基化在其中的调控作用。
      方法 甲状腺乳头状癌细胞系BCPAP和KTC-1细胞用于实验，利用CCK-8法检测碘化钾(KI)对细胞活力的影响。实验分为对照组、碘化钾处理组、碘化钾+DNA甲基化特异性抑制剂地西他滨(decitabine，DAC)处理组和地西他滨处理组。通过EdU细胞增殖检测法检测各组细胞增殖情况；划痕实验检测各组细胞迁移情况；甲基化特异性PCR(MSP)检测各组细胞NIS基因启动子甲基化率；Western blot检测各组细胞NIS蛋白表达情况。
      结果 CCK-8法检测结果显示，1.0×10-3mmol/L KI处理48 h后细胞活性最高，因此选择此浓度进行后续实验。与对照组相比较，KI组的BCPAP细胞和KTC细胞EdU阳性细胞率、划痕愈合率、NIS基因启动子甲基化率显著增加，NIS蛋白表达水平显著降低(P < 0.05)；与KI组相比较，KI+DAC组的BCPAP细胞和KTC细胞EdU阳性细胞率、划痕愈合率、NIS基因启动子甲基化率显著降低，NIS蛋白表达水平显著升高(P < 0.05)。
      结论 高碘可能通过增强NIS基因启动子CpG位点的DNA甲基化水平，促进甲状腺乳头状癌细胞增殖。

       

      Abstract:
      Objective To investigate the effect of high iodine on the development of papillary thyroid carcinoma (PTC) and the regulatory effect of sodium iodide transporter (NIS) gene promoter methylation through an in vitro cell experiment.
      Methods PTC cell lines BCPAP and KTC-1 were selected for the experiment, and the effect of potassium iodide (KI) on cell viability was determined using the CCK-8 method. The experiment included a control group, a KI group, a KI+DNA methylation-specific inhibitor decitabine (DAC) group, and a DAC group. The cell proliferation of each group was determined using EdU cell proliferation assay, and the cell migration using scratch assay. The methylation rate of NIS gene promoter in each group was determined by methylation-specific polymerase chain reaction. The expression of NIS protein in each group was measured by Western blot.
      Results The CCK-8 result showed that the highest cell viability was observed after 48 hours of treatment with 1.0×10-3 mmol/L KI. Therefore, this concentration was chosen for subsequent experiments. Compared with the control group, the EdU positive cell rate, scratch healing rate, and NIS gene promoter methylation rate of BCPAP and KTC cells in the KI group significantly increased, while the NIS protein expression level significantly decreased (P < 0.05). Compared with the KI group, the EdU positive cell rate, scratch healing rate, and NIS gene promoter methylation rate of BCPAP and KTC cells in the KI+DAC group were significantly reduced, while the NIS protein expression level significantly increased (P < 0.05).
      Conclusion High iodine may promote the proliferation of PTC cells by enhancing DNA methylation levels at the CpG site of NIS gene promoter.

       

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