摘要
目的探索垂体肿瘤转化基因结合因子(PBF)调控鼠双微体蛋白2(MDM2)/钠碘转运体(NIS)表达对改善甲状腺癌细胞放射性碘摄取的影响。方法通过慢病毒干预的方式使甲状腺癌细胞TPC-1中PBF蛋白表达降低,分为PBF-shRNA组和对照组,然后在两组TPC-1细胞第1、3、5、7天后进行CCK-8细胞活力检测。加入放射活度为0.185 Bq的Na131I溶液后,对两组TPC-1第1、3、5、7天后进行细胞放射性计数测定。最后通过Western blot实验分析两组TPC-1细胞MDM2、NIS的相对表达量。结果细胞培养1、3、5、7 d后,PBF-shRNA组和对照组的TPC-1细胞计数均呈逐渐增高的生长趋势(F=41.909,P<0.001;F=108.711,P<0.001)。PBF-shRNA组在第5、7天后的细胞活力分别为(59.4±7.1)%和(80.2±5.3)%,均低于对照组的(78.6±6.2)%和(98.5±4.8)%(t=-3.528,P=0.024;t=-4.433,P=0.011)。在细胞培养第5、7天时,PBF-shRNA组的放射性计数分别为(1022.3±80.9)和(1132.2±134.7)Cpm,均高于对照组的(907.5±76.7)和(913.0±69.0)Cpm(t=3.528,P=0.024;t=-4.433,P=0.011)。对照组和PBF-shRNA组在不同时间点细胞活性与放射性计数均呈正相关趋势(r=0.581,P=0.003;r=0.871,P<0.001)。在抑制PBF蛋白表达后,PBF-shRNA组MDM2蛋白相对表达量也低于对照组(t=4.630,P=0.001),但NIS在PBF-shRNA组的蛋白相对表达量高于对照组(t=3.850,P=0.003)。结论TPC-1细胞活力降低和Na131I放射性摄取增高的机制可能与PBF调控MDM2和NIS蛋白表达差异有关。
Objective To investigate the effects of PTTG binding factor(PBF)regulation on murine double minute 2(MDM2)and sodium/iodine symporter(NIS)expression and its impact on improving radioactive iodine uptake in thyroid cancer cells.Methods Lentiviral-mediated silencing of PBF protein expression was performed in TPC-1 thyroid cancer cells,creating two groups:the PBF-shRNA group and the control group.Cell viability was assessed using the CCK-8 assay at days 1,3,5,and 7 after intervention.After exposure to Na131I(radioactive activity:0.185 Bq),radioactive iodine uptake was measured at the same time points.Western blot analysis was used to compare the relative expression of MDM2 and NIS proteins between the two groups.Results Both groups exhibited a gradual increase in TPC-1 cell numbers over 1,3,5,and 7 days of culture(F=41.909,P<0.001;F=108.711,P<0.001).However,cell viability in the PBF-shRNA group was significantly lower than in the control group on days 5 and 7[(59.4±7.1)%and(80.2±5.3)%,respectively,vs.(78.6±6.2)%and(98.5±4.8)%;t=-3.528,P=0.024;t=-4.433,P=0.011].On days 5 and 7,radioactive iodine uptake in the PBF-shRNA group was significantly higher[(1022.3±80.9)Cpm and(1132.2±134.7)Cpm,respectively]compared to the control group[(907.5±76.7)Cpm and(913.0±69.0)Cpm;t=3.528,P=0.024;t=-4.433,P=0.011].A positive correlation was observed between cell viability and radioactive iodine uptake in both the control group and the PBF-shRNA group(r=0.581,P=0.003;r=0.871,P<0.001).Western blot results showed that silencing PBF protein expression reduced MDM2 protein levels(t=4.630,P=0.001)but increased NIS protein expression in the PBF-shRNA group compared to the control group(t=3.850,P=0.003).Conclusion The reduction in TPC-1 cell viability and the increase in Na131I uptake may be associated with PBF regulation of differential MDM2 and NIS protein expression.
作者
夏欢
常诚
吴旻
董占飞
XIA Huan;CHANG Cheng;WU Min;DONG Zhan-fei(Department of Nuclear Medicine,Affiliated Tumor Hospital of Xinjiang Medical University,Urumqi 830011,Xinjiang,China)
出处
《广东医学》
2025年第8期1121-1125,共5页
Guangdong Medical Journal
基金
新疆维吾尔自治区自然科学基金面上项目(2022D01C520)。
关键词
甲状腺癌
甲状腺滤泡上皮细胞
钠/碘转运体
碘
thyroid carcinoma
thyroid follicular epithelial cells
sodium/iodine symporter
iodine
