miR-124通过靶基因IQGAP1调控甲状腺癌细胞分化、增殖的机制

发布时间：2018-06-23 04:53

本文选题：甲状腺癌 + miRNA　；参考：《中国老年学杂志》2017年03期

【摘要】：目的探讨miR-124对甲状腺癌细胞的作用机制。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-124的表达水平。MTT法检测甲状腺癌细胞K1转染miR-124 mimics、mimics control后的细胞存活率。根据靶基因预测软件The miRBase、Target Scan、Pic Tar预测miR-124的靶基因IQGAP1,构建wt-IQGAP1和mut-IQGAP1,采用双荧光素酶活性检测鉴定靶基因的正确性。Western印迹检测转染miR-124 mimics、mimics control后甲状腺癌细胞K1中IQGAP1的表达情况。甲状腺癌细胞K1中转染si IQGAP1和si IQGAP1 control,用MTT检测细胞增殖能力,Western印迹检测细胞中IQGAP1蛋白表达含量。结果甲状腺癌细胞K1、BCPAP、TPC-1中的miR-124的相对表达量与甲状腺细胞相比差异显著(P0.05),三种甲状腺癌细胞中K1细胞的表达量最低。转染后12 h内,空白组、miR-124 mimics组和mimics control组的甲状腺癌K1细胞的存活数量差异不显著(P0.05),从转染后12 h开始,miR-124 mimics组与对照组、mimics control组相比,甲状腺癌细胞生长抑制明显。预测miR-124的靶基因可能为IQGAP1,转染miR-124 mimics野生型IQGAP1中荧光素酶活性比突变型IQGAP1中荧光素酶活性显著降低,差异显著(P0.05)。mimics control和wt-IQGAP1和mut-IQGAP1共转染组比较无明显差异(P0.05)。转染miR-124 mimics组甲状腺癌K1细胞IQGAP1蛋白明显下降,miR-124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-124在甲状腺癌细胞中表达下调,miR-124可以抑制甲状腺癌细胞的增殖分化,其通过负调控靶基因IQGAP1抑制癌细胞增殖分化。
[Abstract]:Objective to investigate the mechanism of miR-124 acting on thyroid cancer cells. Methods the expression level of miR-124 in human thyroid cancer cell K1 / BCPAPP / TPC-1 and human thyroid cell Nthy-ori 3-1 was detected by RT-PCR. The survival rate of K1 transfected with miR-124 mimicsmimics control was detected by MTT assay. The target gene IQGAP1 of miR-124 was predicted by the target gene prediction software, the miRBaseScanTar. Wt-IQGAP1 and mut-IQGAP1 were constructed. The expression of IQGAP1 in thyroid cancer cell K1 after transfection of miR-124 mimicsmimics control was detected by double luciferase activity assay. Si IQGAP1 and si IQGAP1 control1 were transfected into thyroid cancer cell K1 and the expression of IQGAP1 protein was detected by MTT assay and Western blotting. Results the relative expression of miR-124 in thyroid cancer cell K1 BCPAPAPP TPC-1 was significantly different from that in thyroid cell (P0.05), and the expression of K1 in three kinds of thyroid cancer cells was the lowest. Within 12 hours after transfection, there was no significant difference in the survival of K1 cells between the blank group and the mimics control group (P0.05). From 12 h after transfection, the growth inhibition of thyroid cancer cells in the miR-124 mimics group was significantly higher than that in the control group (mimics control group). It was predicted that the target gene of miR-124 might be IQGAP1. The luciferase activity of transfected miR-124 mimics wild-type IQGAP1 was significantly lower than that of mutant IQGAP1 (P0.05). There was no significant difference between mimics control and wt-IQGAP1 and mut-IQGAP1 co-transfection groups (P0.05). The expression of IQGAP1 in K1 cells of thyroid carcinoma was significantly decreased by miR-124 mimics transfection, and the expression of IQGAP1 was negatively regulated by miR-124. Inhibition of IQGAP1 gene in K1 cells decreased cell proliferation and IQGAP1 protein expression. Conclusion the down-regulated expression of miR-124 in thyroid cancer cells can inhibit the proliferation and differentiation of thyroid cancer cells, and it can inhibit the proliferation and differentiation of thyroid cancer cells through negative regulation target gene IQGAP1.
【作者单位】：天津市海河医院内科;天津市医科大学总医院内分泌科;
【分类号】：R736.1

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