miR-146b调控甲状腺乳头状癌细胞生物学行为的机制研究  

Mechanism of miRNA-146b regulating proliferation, metastasis and apoptosis of thyroid papillary carcinoma cells

作　　者：方燕 郑旭旭 李丽燕[1] Fang Yan;Zheng Xuxu;Li Liyan(Department of Pathology,Wenzhou Hospital of Traditional Chinese Medicine,Wenzhou 325000,China)

机构地区：[1]温州市中医院病理科,温州325000

出　　处：《中华内分泌外科杂志》2022年第1期58-63,共6页Chinese Journal of Endocrine Surgery

基　　金：温州市科技计划项目 (Y20190419)。

摘　　要：目的探讨miR-146b在甲状腺乳头状癌(papillary thyroid carcinoma,PTC)细胞增殖、转移和凋亡中的作用及相关机制。方法qRT-PCR检测miR-146b在PTC细胞(NPA、GLAG-66、ONCNO-DG1和B-CPAP)和正常人甲状腺细胞系HTori3中的表达水平。miR-146b抑制剂转染B-CPAP细胞后,通过qRT-PCR检测其抑制效率,利用MTT实验、Transwell实验和流式细胞术分别检测miR-146b对PTC细胞增殖、侵袭能力及凋亡率的影响。SiRNA-IRAK1转染B-CPAP细胞后,利用MTT实验、细胞划痕实验和流式细胞术分别检测IRAK1对PTC细胞增殖率、转移及凋亡率的影响。利用生物信息学软件预测miR-146b的靶基因白介素1关联受体激酶1(interleukin-1 receptor-associated kinases,IRAK1),并通过双荧光素报告基因实验验证miR-146b对IRAK1的调控作用。应用SPSS 20.0软件进行分析,组间对比采用两独立样本t检验。结果qRT-PCR结果表明,与正常细胞HTori3相比,miR-146b在NPA87、KAT-5、FTC-133和B-CPAP细胞系中表达显著升高,尤以B-CPAP较高[相对表达量1 vs(1.61±0.11)、(1.92±0.21)、(1.93±0.22)、(2.43±0.31),P<0.05]。miR-146b抑制剂转染后可显著降低B-CPAP细胞中miR-146b的表达水平[相对表达量1 vs(0.41±0.12,P<0.01]。MTT结果发现,miR-146b抑制剂可抑制B-CPAP细胞的增殖能力[相对增殖率值1.00 vs(0.52±0.12)、(0.56±0.11)、(0.62±0.12),P<0.05]。流式细胞仪检测发现,miR-146b抑制剂可促进B-CPAP细胞的凋亡能力[(3.74%±0.12%)vs(7.62%±0.21%),P<0.05]。Transwell结果表明,miR-146b抑制剂可减弱B-CPAP细胞的侵袭能力[侵袭细胞数目(280.00±67.00)vs(82.00±32.00),P<0.05]。转染入siRNA-IRAK1后,B-CPAP细胞增殖率显著升高[MTT实验,相对增殖率1 vs(2.11±0.21)、(2.33±0.18)、(2.21±0.16)]、转移能力增强[细胞划痕实验,24 h相对迁移率(0.58%±0.11%)vs(0.81%±0.21%)]且细胞凋亡率显著降低[流式细胞术,相对凋亡率(6.96%±1.12%)vs(24.30±1.52%),P<0.05]。双荧光素酶报告基因结果显示,IRAK1Objective To investigate the role and mechanism of miR-146b in the proliferation,metastasis and apoptosis of thyroid papillary carcinoma cells.Methods qRT-PCR was used to detect the expression of miR-146b in thyroid papillary carcinoma cells(NPA,GLAG-66,ONCNO-DG1 and B-CPAP)and normal human thyroid cell line HTori3.After B-CPAP cells were transfected with miR-146b inhibitor,the inhibition efficiency was detected by qRT-PCR,the effect of miR-146b on PTC cells proliferation was detected by MTT assay,the effect of miR-146b on PTC cells invasion was studied by Transwell assay,and the effect of miR-146b on tumor cell apoptosis was detected by flow cytometry.SiRNA-IRAK1 was transfected into B-CPAP cell line.The cell proliferation rate,migration ability and apoptosis rate were detected by MTT,cell scratch test and flow cytometry respectively.The target gene of miR-146b,interleukin-1 associated receptor kinase 1(IRAK1),was predicted by bioinformatics software,and the regulatory effect of miR-146b on IRAK1 was verified by double fluorescein reporter gene experiment.Results QRT-PCR showed that the expression of miR-146b in NPA87,KAT-5,FTC-133 and B-CPAP cell lines was significantly higher than that in normal cell HTori3,especially B-CPAP(P<0.05).MiR-146b inhibitor transfection could significantly reduce the expression level of miR-146b in B-CPAP cells(P<0.01).MTT results showed that miR-146b inhibitor could inhibit the proliferation of B-CPAP cells(P<0.05).Flow cytometry showed that miR-146b inhibitor could promote the apoptosis of B-CPAP cells(P<0.05).Transwell results showed that miR-146b inhibitor could reduce the invasive ability of B-CPAP cells(P<0.05).After transfection with siRNA-IRAK1,the proliferation rate of B-CPAP cells increased significantly(MTT test),the migration ability increased(cell scratch test),and the apoptosis rate decreased significantly(flow cytometry)(P<0.05).The results of double luciferase reporter gene showed that irak1 was the target gene of miR-146b,and miR-146b inhibitor could significantly u

关 键 词：甲状腺乳头状癌 miR-146b 转移 靶基因白介素1关联受体激酶1 

分 类 号：R653[医药卫生—外科学] R736.1[医药卫生—临床医学]