微RNA-21-5p靶向调控含硬化蛋白域蛋白1对甲状腺癌细胞生长的影响  

Impact of microRNA-21-5p on the growth of thyroid cancer cells via targeting the recombinant sclerostin domain containing protein 1

作　　者：刘妙妙[1] 邓豪余[1] 赵雅洁[1] 李灿[1] 刘桦[1] LIU Miaomiao;DENG Haoyu;ZHAO Yajie;LI Can;LIU Hua(Department of Nuclear Medicine,Xiangya Hospital,Central South University,Changsha 410008,China)

机构地区：[1]中南大学湘雅医院核医学科,长沙410008

出　　处：《中南大学学报（医学版）》2021年第10期1054-1062,共9页Journal of Central South University ：Medical Science

摘　　要：目的:通过分析微RNA(microRNA,miR)-21-5p及其靶基因含硬化蛋白域蛋白1(recombinant sclerostin domain containing protein 1,SOSTDC1)在甲状腺癌中的作用,深入了解甲状腺癌转移的分子机制。方法:通过生物信息学分析和细胞验证筛选出miR-21-5p,通过miR-21-5p抑制剂转染甲状腺癌细胞系;采用MTT实验、流式细胞术和细胞划痕实验分别检测miR-21-5p抑制剂组和抑制剂对照组的甲状腺癌细胞增殖、凋亡和迁移的情况;采用荧光素酶报告实验验证miR-21-5p和SOSTDC1的靶向调控关系;采用蛋白质印迹法检测miR-21-5p抑制剂组和抑制剂对照组甲状腺癌细胞中SOSTDC1、下游磷脂酰肌醇3-激酶(phosphatidylinositol 3 kinase,PI3K)/蛋白激酶B(protein kinase B,Akt)和丝裂原活化蛋白激酶(mitogen-activated protein kinases,MAPK)/细胞外调节蛋白激酶(extracellular regulated protein kinases,ERK)信号通路因子的表达水平及磷酸化水平。结果:MiR-21-5p在甲状腺癌细胞中显著上升,且与SOSTDC1呈负相关(r=-0.24,P<0.01);miR-21-5p抑制剂组甲状腺癌细胞增殖和迁移显著低于抑制剂对照组(均P<0.01),细胞凋亡率显著高于抑制剂对照组(P<0.01);荧光素酶报告实验表明miR-21-5p能够靶向调控SOSTDC1表达水平;甲状腺癌细胞中PI3K/Akt和MAPK/ERK信号通路检测显示miR-21-5p抑制剂组PI3K表达水平显著低于抑制剂对照组(P<0.01),Akt和ERK1/2水平无显著变化,但miR-21-5p抑制剂组Akt和ERK1/2磷酸化水平显著低于抑制剂对照组(均P<0.01)。结论:甲状腺癌细胞中miR-21-5p能够靶向抑制SOSTDC1的表达,影响PI3K/Akt和MAPK/ERK活性,从而抑制甲状腺癌细胞凋亡,促进细胞的增殖和迁移能力。Objective: To explore the molecular mechanism for thyroid cancer metastasis via analyzing the role of micro RNA(mi R)-21-5 p and its target gene recombinant sclerostin domain containing protein 1(SOSTDC1) in thyroid cancer.Methods: The target miR-21-5p was screened through bioinformatics analysis and cell verification, and the thyroid cancer cell lines was transfected with miR-21-5p inhibitor. 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide(MTT) test, flow cytometry,and cell scratch test were used to detect the proliferation, apoptosis and migration of thyroid cancer cells in the miR-21-5p inhibitor group and the inhibitor control group,respectively. The luciferase report experiment was used to verify the relationship between miR-21-5p and SOSTDC1, Western blotting was used to detect the expression levels and phosphorylation levels of SOSTDC1, phosphatidylinositol 3 kinase(PI3K), protein kinase B(Akt) and mitogen-activated protein kinases(MAPK), extracellular regulated protein kinases(ERK) in thyroid cancer cells.Results: MiR-21-5p was significantly increased in thyroid cancer cells, which was negatively correlated with SOSTDC1(r=-0.24, P<0.01). The proliferation and migration of thyroid cancer cells in the miR-21-5p inhibitor group was significantly lower than that in the inhibitor control group(both P<0.01), and the apoptosis rate in the miR-21-5p inhibitor group was significantly higher than that in the inhibitor control group(P<0.01). The luciferase report experiment showed that miR-21-5p could target and regulate the expression level of SOSTDC1, and the expression of PI3K in the miR-21-5p inhibitor group was significantly lower than that in the inhibitor control group(P<0.01). There were no significant changes in Akt and ERK1/2 levels, but the phosphorylation levels of Akt and ERK1/2 in the miR-21-5p inhibitor group were significantly lower than those in the inhibitor control group(both P<0.01).Conclusion: MiR-21-5p in thyroid cancer cells can target the expression of SOSTDC1 and affect the

关 键 词：甲状腺癌 微RNA-21-5p 含硬化蛋白域蛋白1 

分 类 号：R736.1[医药卫生—肿瘤]