miR-142影响甲状腺癌细胞增殖分化的机制研究  

The Role of miR-142 in Proliferation and Differentiation of Thyroid Cancer Cell

作　　者：刘春梅 孙小霞 杨亚峰 潘娟 宋宗良[2] 宗伟[3] 

机构地区：[1]咸阳市中心医院内分泌科,陕西咸阳712000 [2]陕西中医药大学附属医院内分泌科,陕西咸阳712000 [3]陕西省人民医院消化内科,陕西西安710000

出　　处：《肿瘤药学》2017年第1期17-22,共6页Anti-Tumor Pharmacy

基　　金：陕西省自然科学基础研究计划项目(2014JM4141)

摘　　要：目的探讨miR-142对甲状腺癌细胞增殖与分化的作用机制。方法通过RT-PCR检测人甲状腺癌细胞K1、BCPAP、TPC-1和人甲状腺细胞Nthy-ori 3-1中miR-142的表达水平。细胞分别转染miR-142 mimics si IQGAP。MTT法检测细胞存活率与增殖能力。构建wt-IQGAP1和mut-IQGAP1载体,利用双荧光素酶活性检测鉴定靶基因。Western blot检测甲状腺癌细胞K1中IQGAP1的表达情况。结果甲状腺癌细胞K1、BCPAP、TPC-1中的miR-142相对表达量与甲状腺细胞相比显著降低(P<0.05),三种甲状腺癌细胞中K1细胞的表达量最低。从转染后12 h开始,miR-142 mimics组与空白组、mimics control组相比,甲状腺癌细胞生长抑制明显。预测miR-142的靶基因可能为IQGAP1,转染miR-142 mimics野生型IQGAP1中荧光素酶活性较突变型IQGAP1中荧光素酶活性显著降低(P<0.05)。转染miR-142 mimics组甲状腺癌K1细胞IQGAP1蛋白明显下降,miR-142负调控IQGAP1的表达。抑制甲状腺癌K1细胞中IQGAP1基因,细胞增殖能力减弱,细胞中IQGAP1蛋白表达减弱。结论 miR-142在甲状腺癌细胞中表达下调,miR-142可以通过负调控靶基因IQGAP1抑制甲状腺癌细胞的增殖分化。Objective To evaluate the influence of miR-142 on the proliferation and differentiation of thyroid cancer cell. Methods The expression levels of miR-142 in thyroid cancer ceils K1, BCPAP, TPC-1 and human thyroid cells Nthy-ori 3-1 were detected by RT- PCR. miR-142 mimics and siIQGAP were transfected in the cancer cells, respectively. MTT assay was used to detect cell proliferation and survival rate. Wt-IQGAP1 and mut-IQGAP1 were constructed, and dual luciferase activity was used for identification of target genes. The IQGAP1 expression was detected by western blot in thyroid cancer cells K1. Results Compared with thyroid cells, the relative expressions of miR-142 in thyroid cancer cells K1, BCPAP and TPC-1 were significantly decreased （P〈0.05）. The expression of miR-142 in thyroid cancer K1 cells was lowest. After 12 h transfection, compared with the control group and mimics control group, the thyroid cancer cell growth was significantly inhibited in miR-142 mimics group. IQGAP1 was predicted to be the target gene of miR-142. Compared with transfection of miR-142 mimics and mut-IQGAP1 group, the luciferase activity was significantly decreased in the miR-142 mimics and wt-IQGAP1 group （P〈0.05）. The IQGAP1 expression in thyroid cancer K1 cells was decreased significantly in miR-142 mimics group, indicating that miR-142 negatively regulated the expression of IQGAP1. Conclusions miR-142 was down-regulated in thyroid cancer cells, miR-142 can proliferation and differentiation of inhibit thyroid cancer cell by negatively regulating target genes IQGAP1.

关 键 词：甲状腺癌 MIRNA 靶基因 增殖 

分 类 号：R736.1[医药卫生—肿瘤]