摘要
目的探讨线粒体自噬诱导剂GSK3-IN-3在增强甲状腺癌细胞对放射性碘-131治疗敏感性的潜在作用。方法通过不同浓度GSK3-IN-3处理甲状腺癌细胞BCPAP,采用RT-PCR和Western blot检测钠碘同向转运体(NIS)mRNA及蛋白的表达水平;使用免疫荧光观察NIS蛋白的细胞膜定位;使用γ计数器测定碘-131的摄取量;通过CCK-8检测细胞增殖情况;免疫荧光技术评估细胞线粒体自噬水平。结果GSK3-IN-3增加了甲状腺癌细胞中NIS mRNA和蛋白的表达水平(P<0.001),并增强了NIS蛋白在细胞膜上的定位,且随着GSK3-IN-3浓度增加而增强。此外,25μM GSK3-IN-3处理后,细胞对碘-131的摄取量提高(P<0.001),在重组甲状腺刺激激素(rTSH)刺激下摄取量进一步增加;加入NIS特异性抑制剂NaClO4后,碘-131摄取量下降(P<0.001)。CCK-8检测结果显示,GSK3-IN-3处理组的细胞增殖率降低(P<0.05)。免疫荧光观察表明,GSK3-IN-3处理后甲状腺癌细胞的线粒体自噬水平提升。结论GSK3-IN-3能有效促进BCPAP甲状腺癌细胞中NIS mRNA和蛋白的表达,增强碘-131的摄取,并有效抑制细胞增殖,为甲状腺癌放射性碘-131治疗提供了新的机制理解。
Objective To investigate the potential role of the mitochondrial autophagy inducer GSK3-IN-3 in enhancing the sensitivity of thyroid cancer cells to radioactive iodine-131(I-131)treatment.Methods Thyroid cancer BCPAP cells were treated with different concentrations of GSK3-IN-3.The expression levels of Sodium Iodide Symporter(NIS)mRNA and protein were detected using RT-PCR and Western blotting.Immunofluorescence was used to observe the membrane localization of NIS protein.The uptake of I-131 was measured using a gamma counter.Cell proliferation was assessed using the CCK-8 assay.Mitochondrial autophagy levels in cells were evaluated using immunofluorescence techniques.Results GSK3-IN-3 increased the expression levels of NIS mRNA and protein in thyroid cancer cells(P<0.001),and enhanced the membrane localization of NIS protein,with this effect being concentration-dependent.Additionally,treatment with 25μM GSK3-IN-3 significantly increased I-131 uptake(P<0.001),which was further enhanced by recombinant Thyroid-Stimulating Hormone(rTSH)stimulation.The I-131 uptake was significantly reduced when the NIS-specific inhibitor NaClO4 was added(P<0.001).The CCK-8 assay showed that cell proliferation in the GSK3-IN-3 treated group was significantly reduced(P<0.05).Immunofluorescence observations indicated that the mitochondrial autophagy level in thyroid cancer cells was significantly elevated after GSK3-IN-3 treatment.Conclusion GSK3-IN-3 effectively promotes the expression of NIS mRNA and protein in BCPAP thyroid cancer cells,enhances I-131 uptake,and significantly inhibits cell proliferation,providing new mechanistic insights into the use of radioactive iodine-131 treatment in thyroid cancer.
作者
陈卓
张玉娇
杨国栋
叶佳
张平
罗高潮
陈文平
CHEN Zhuo;ZHANG Yujiao;YANG Guodong;YE Jia;ZHANG Ping;LUO Gaochao;CHEN Wenping(Department of Nuclear Medicine,Huanggang Central Hospital Affiliated to Yangtze University,Huanggang 438000,Hubei,China;Oncology Center,Huanggang Central Hospital Affiliated to Yangtze University,Huanggang 438000,Hubei,China;Department of Endocrinology,Rheumatology and Immunology,Huanggang Central Hospital Affiliated to Yangtze University,Huanggang 438000,Hubei,China)
出处
《西部医学》
2025年第8期1129-1134,共6页
Medical Journal of West China
基金
湖南省人民医院医联体专项科研基金项目(2022YLT005)。
