摘要
目的:研究miR-194、EZH2对甲状腺癌细胞增殖、迁移和侵袭的影响,并探讨其作用机制。方法:采用qRT-PCR法检测甲状腺癌组织、癌旁正常组织及甲状腺癌细胞和正常甲状腺上皮细胞中miR-194和EZH2的mRNA表达;将miR-194(转染miR-194 mimics)、miR-NC(未转染细胞)、inhibitor-NC(转染空inhibitor)、miR-194 inhibitor(转染miR-194 inhibitor)、si-EZH2(转染si-EZH2)、miR-194+Vector(miR-194 mimics和pcDNA 3.1共转染)、miR-194+EZH2(miR-194 mimics和pcDNA 3.1-EZH2共转染)均以脂质体法转染到SW579、IHH-4细胞;Western blot检测细胞中EZH2的蛋白表达;MTT法检测细胞的增殖;Transwell检测细胞的迁移和侵袭;双荧光素酶报告基因检测实验检测细胞荧光素酶活性。结果:与正常组相比,甲状腺癌组EZH2的表达显著升高,miR-194的表达显著降低;与人正常甲状腺上皮细胞相比,甲状腺癌细胞中EZH2表达显著升高,miR-194的表达显著降低,且过表达miR-194和沉默EZH2均可抑制甲状腺癌细胞增殖、迁移和侵袭。EZH2为miR-194的靶标,且EZH2可逆转过表达miR-194对甲状腺癌细胞增殖、迁移和侵袭的抑制作用。结论:miR-194可抑制甲状腺癌细胞的增殖、迁移和侵袭,可能与靶向EZH2有关,将可为miR-194靶向治疗甲状腺癌提供依据。
Objective:To study the effects of miR-194 and EZH2 on cell proliferation,migration and invasion in thyroid cancer cells,and explore its mechanism.Methods:qRT-PCR was used to detect mRNA expression of miR-194 and EZH2 in tissues and cells.miR-194(transfected miR-194 mimics),miR-NC(untransfected cells),inhibitor-NC(transfected empty inhibitor),miR-194 inhibitor(transfected miR-194 inhibitor),si-EZH2(transfected si-EZH2),miR-194+Vector(miR-194 mimics and pcDNA 3.1 co-transfection),miR-194+EZH2(miR-194 mimics and pcDNA 3.1-EZH2 co-transfection) were transfected into SW579 and IHH-4 cells by liposome method.Western blot was used to detect the protein expression of EZH2 in cells.MTT assay was used to detect cell proliferation.Transwell was used to detect cell migration and invasion.Dual luciferase reporter assay was used to detect luciferase activity.Results:Compared with the normal thyroid group,the expression of EZH2 was significantly increased in thyroid cancer group,and the expression of miR-194 was significantly decreased.Compared with normal thyroid epithelial cells,the expression of EZH2 was significantly increased in thyroid cancer cells and miR-194 was significantly decreased,and overexpression miR-194 or silencing EZH2 inhibited cell proliferation,migration and invasion in thyroid cancer cells.EZH2 is a target of miR-194.EZH2 could rescue the inhibitory effect of overexpression miR-194 on cell proliferation,migration and invasion in thyroid cancer cells.Conclusion:miR-194 could inhibits the proliferation,migration and invasion of thyroid cancer cells.It may be related to targeting EZH2,which will provide a basis for miR-194 targeted therapy for thyroid cancer.
作者
刘兵雄
宋艾红
李学刚
Liu Bingxiong;Song Aihong;Li Xuegang(The Third Department of General Surgery of Hanchuan People's Hospital,Hubei Hanchuan 431600,China)
出处
《现代肿瘤医学》
CAS
2020年第7期1101-1107,共7页
Journal of Modern Oncology
