微小RNA-451靶向巨噬细胞游走抑制因子调控甲状腺癌细胞增殖、凋亡的机制  

The mechanism of microRNA-451 targeting macrophage migration inhibitory factor on prolifera- tion and apoptosis of thyroid cancer cells

作　　者：李晓娜[1] 李玉杰[1] 董玉科[1] 于敏[1] 黄炜[1] 李冬枝[1] 赵春红[1] 孙旭[1] 李韬[1] 邢国臣[2] 闫巧辉[2] 

机构地区：[1]郑州大学附属郑州中心医院头颈外科,450007 [2]郑州大学附属郑州中心医院肿瘤科,450007

出　　处：《中华实验外科杂志》2017年第6期956-958,共3页Chinese Journal of Experimental Surgery

摘　　要：目的探讨微小RAN（miR）-451靶向巨噬细胞游走抑制因子（MIF）对甲状腺癌细胞增殖、凋亡的影响及机制。方法反转录．聚合酶链反应（RT—PCR）检测甲状腺癌组织及滤泡状（PTC-133）、乳头状（K1）及未分化（8505C）甲状腺癌细胞株中miR-451的mRNA表达；miR—NC、miR-451模拟物（mimics）、miR-451抑制物（inhibitor）转染K1甲状腺癌细胞，Western blot检测MIF蛋白表达，3组共转染组miR-NC＋miR-451、Wt—MIF＋miR-451mimics、Mut-MIF＋miR-451mimics转染K1甲状腺癌细胞，检测荧光素酶活性，以此证实MIF是否为miR-451靶基因；后续实验分为miR—NC、miR-451mimics、miR-451mimics＋pcDNA3．1-MIF及miR—NC＋pcDNA3．1-MIF4组，各组细胞培养48h后，细胞计数试剂盒（CCK-8）实验检测细胞增殖，流式细胞仪检测细胞凋亡，Western blot检测活化的含半胱氨酸的天冬氨酸蛋白水解酶-3（Cleaved Caspase-3）、磷脂酰肌醇-3-激酶（P13K）、蛋白激酶B（Akt）、磷酸化蛋白激酶B（p-Akt）蛋白表达。结果甲状腺癌组织中miR-451的mRNA表达（0．889±0．367）显著低于癌旁组织（6．214±0．884）（t=28．883，P=0．004），甲状腺癌细胞中K1细胞的表达最低，选择作为后续研究；MIF是miR-451的靶基因；与miR-NC组比较P13K（0．562±0．051）；p-Akt（0．134±．022）；CleavedCaspase-3（0．052±0．014）；细胞凋亡率（5．16±0．77）％，miR-451mimics和nliR-451mimics＋pcDNA3．1-MIF组P13K（0．133±0．021、0．223±0．026）、p—Akt（0．034±0．011、0．062±0．013）蛋白表达显著降低（与miR-451mimics组比较：tPI3K=30．157，P=0．003；tp-Akt=46．626，P=0．001。结论MIF是miR-451的靶基因，miR-451可通过靶向MIF抑制P13K／Akt信号通路抑制K1甲状腺癌细胞增殖及促进细胞凋亡。Objective To investigate the effect and mechanism of microRNA （miR） -451 targe- ting macrophage migration inhibitory factor on proliferation and apoptosis of thyroid cancer cells. Methods MiR -451 mRNA expression in thyroid carcinoma and follicular papillary （PTC -133 ） , （Kl） and undif- ferentiated （8505C） thyroid cancer cell lines were detected by reverse transcription -polymerase chain re- action （RT- PCR）; miR- NC, miR- 451 mimics （Analog）, miR-451 inhibitor （inhibitor） were trans- fected with thyroid cancer K1 ceils, and macrophage migration inhibitory （MIF） protein expression were detected by Western blotting, the three Co transfection group miR - NC ＋ miR - 451 mimics, Wt-MIF ＋miR-451 mimics, Mut- MIF ＋ miR- 200b mimics was transfected into K1 thyroid cancer ceils, luciferase activity was detected to confirm whether MIF is the target gene of miR -451 ; the follow - up experiment was divided into miR- NC, miR-451 mimics, miR- 451 mimics ＋ pcDNA3.1 -MIF and miR - NC ＋ pcDNA3. 1 - MIF, each group cell were cultured for 48 h, cell proliferation was detected by cell counting kit （ CCK - 8） test; apoptosis was detected by flow cytometry; the expression of Cleaved Caspase-3, phosphatidylinositol 3 kinase （PI3K）, protein kinase B （Akt）, phosphorylated protein ki- nase B （p -Akt） protein was detected by Western blotting. Results The expression of miR -451 mRNA （0. 889 ± 0. 367 ） in thyroid carcinoma was significantly lower than adjacent tissues （ 6. 214 ± 0. 884 ）, （ t = 28. 883, P = 0.004 ） , the lowest expression of K1 cells in thyroid cancer cells, selected as a follow - up study; compared with miR - NC group [ PI3K （0. 562 ± 0. 051 ） p - Akt （0. 134 ± 0. 022） Cleaved Caspase - 3 （0. 052 ± 0. 014 ） cell apoptosis rate （5.16 ± 0. 77 ） %, PI3K （0. 133 ± 0. 021, 0. 223 ± 0. 026） , p - Akt （ 0.034 ± 0. 011, 0. 062 ± 0. 013 ） protein expression in miR - 451 mimics and miR -451 mimics ＋ peDNA3. 1 -MIF group was si

关 键 词：微小RNA-451 巨噬细胞游走抑制因子 甲状腺癌 增殖 凋亡 磷脂酰肌醇3-激酶/蛋白激酶B信号通路 

分 类 号：R736.1[医药卫生—肿瘤]