溴结构域蛋白4经PTEN/PI3K/AKT通路对甲状腺乳头状癌细胞摄碘相关指标影响的研究  

Study on the effect of bromodomain-containing protein 4 via PTEN/PI3K/AKT pathway on iodine uptake related indicators in human papillary carcinoma of thyroid cells

作　　者：刘帆[1,2] 张雅兰 柴雅晖 宋爱琳[1,2] Liu Fan;Zhang Yalan;Chai Yahui;Song Ailin(The Second Clinical Medical School,Lanzhou University,Lanzhou 730030,China;Department of Greneral Surgery,The Second Hospital of Lanzhou University,Lanzhou 730030,China)

机构地区：[1]兰州大学第二临床医学院,甘肃兰州730030 [2]兰州大学第二医院普通外科,甘肃兰州730030

出　　处：《兰州大学学报（医学版）》2023年第2期6-12,19,共8页Journal of Lanzhou University（Medical Sciences）

基　　金：甘肃省自然科学基金资助项目(20JR5RA342,1JR11RA109)。

摘　　要：目的研究溴结构域蛋白4(BRD4)在甲状腺乳头状癌(PCT)中的表达水平及其抑制剂JQ1对PCT细胞增殖、迁移能力的影响,进一步探索影响摄碘能力的调控机制。方法采用实时荧光定量聚合酶链反应(qRT-PCR)技术及蛋白质印迹法分别检测BRD4在正常甲状腺细胞Nthy-ori 3-1及人PCT细胞TPC-1、K1的表达水平。采用CCK-8及划痕实验分别检测JQ1对TPC-1、K1细胞增殖及迁移能力的影响。采用高内涵细胞成像与分析系统观察TPC-1、K1细胞在JQ1作用下的形态及数目变化。采用qRT-PCR技术检测影响PCT摄碘能力相关基因的表达。结果BRD4在PCT细胞及癌组织中高表达。JQ1对TPC-1、K1细胞增殖能力呈时间及浓度依赖性抑制。TPC-1、K1细胞的48 h半抑制浓度分别为(9.045±0.772)、(14.169±1.406)μmol/L。JQ1可以抑制TPC-1、K1细胞的迁移,且具有统计学意义(P<0.05)。JQ1分别作用于TPC-1、K1细胞48 h后,实验组BRD4、磷脂酰肌醇3激酶(PI3K)和蛋白激酶B(AKT)的mRNA表达水平降低。第10号染色体同源缺失性磷酸酶-张力蛋白基因(PTEN)、钠-碘转运蛋白的mRNA表达升高。结论BRD4在PCT中呈现高表达状态。JQ1可有效抑制BRD4的表达,并影响PTEN/PI3K/AKT通路从而抑制PCT细胞的增殖及迁移,使得其摄碘能力增加。Objective To investigate the expression level of bromodomain protein 4(BRD4)in papillary carcinoma of thyroid(PCT)and the effect of its inhibitor JQ1 on the proliferation and migration ability of PCT,and to further explore the regulatory mechanisms affecting the iodine uptake ability.Methods The BRD4 expression status in Nthy-ori 3-1 of normal thyroid cells and TPC-1 and K1 of human PCT cells was tested by real-time quantitative PCR(qRT-PCR)and Western Blotting technology,respectively.The impact of JQ1 upon the proliferation and migration ability of TPC-1 and K1 cells was tested by CCK-8 and scratch assay.Cell morphology and number were observed using the high-content cell imaging and analysis system.The expression of relevant genes affecting the iodine uptake capacity of PCT was investigated by means of qRT-PCR.Results BRD4 was highly expressed in PCT cells and cancer tissues.JQ1 showed time and concentration-dependent inhibition of the proliferation ability of TPC-1 and K1 cells.The half-inhibitory concentrations(IC50)of TPC-1 cells and K1 cells for 48h were(9.045±0.772)μmol/L and(14.169±1.406)μmol/L,respectively.JQ1 could inhibit the migration of TPC-1 and K1 cells with statistical significance(P<0.05).After JQ1 was applied to TPC-1 and K1 cells for 48h,the mRNAs of BRD4,phosphoinositide 3-kinase(PI3K)and protein kinase B(AKT)were decreased in the experimental group.The mRNAs of phosphatase and tension homology deleted on chromosome ten(PTEN)and sodium iodide symporter(NIS)were increased.Conclusion BRD4 was highly expressed in PCT.JQ1 effectively inhibited the expression of BRD4 and affected the PTEN/PI3K/AKT pathway,thereby inhibiting the proliferation and migration of PCT cells and resulting in increased iodine uptake.

关 键 词：溴结构域蛋白4 甲状腺乳头状癌 磷脂酰肌醇3激酶 蛋白激酶B 钠-碘转运蛋白 

分 类 号：R736.1[医药卫生—肿瘤]