Hsa_circ_0016600通过调控miR-149/PTK7轴促进甲状腺癌细胞迁移的机制

The mechanism of hsa_circ_0016600 promoting thyroid cancer cell migration via modulating the miR-149/PTK7 axis

目的:探究环状RNA hsa_circ_0016600对甲状腺癌细胞迁移的影响及其作用机制。方法:收集温州医科大学附属第二医院颈部外科收治的15例甲状腺癌患者的癌组织及正常组织标本,采用RT-qPCR法检测hsa_circ_0016600、miR-149和PTK7在甲状腺癌组织及甲状腺细胞系中的表达情况。体外培养甲状腺癌细胞系,分别敲低hsa_circ_0016600和过表达miR-149,采用CCK-8、Transwell和划痕实验检测甲状腺细胞增殖、迁移和侵袭能力。建立小鼠异种移植皮下瘤模型,观察敲低hsa_circ_0016600的甲状腺癌细胞在体内的成瘤作用,并利用荧光原位杂交实验检测hsa_circ_0016600在裸鼠移植瘤模型中的表达水平。采用RNA荧光原位杂交实验检测hsa_circ_0016600和miR-149在甲状腺癌细胞中的定位,双荧光素酶报告基因实验检测hsa_circ_0016600与miR-149及miR-149与PTK7的靶向调控关系。结果:在人甲状腺癌组织和细胞系中,hsa_circ_0016600(P<0.05)和PTK7表达上调(P<0.01),miR-149表达下调(P<0.05)。敲低hsa_circ_0016600表达后,甲状腺癌细胞的增殖(P<0.01)、侵袭(P<0.01)和迁移能力(P<0.01)受到抑制。hsa_circ_0016600可通过靶向miR-149促进甲状腺癌细胞的增殖(P<0.01)、迁移(P<0.01)和血管形成,并上调PTK7的表达(P<0.01)。此外,PTK7是miR-149的一个靶基因。结论:Hsa_circ_0016600在甲状腺癌组织和细胞中高表达,并通过调控miR-149/PTK7轴促进甲状腺癌细胞的迁移。

Objective:To investigate the impact of circular RNA hsa_circ_0016600 on the migration of thyroid cancer cells and its mechanisms.Methods:Cancer tissues and normal tissue specimens from 15 thyroid cancer patients treated at the Department of Pate Surgery,the Second Affiliated Hospital of Wenzhou Medical University.Utilize RT-qPCR to assess the expression levels of hsa_circ_0016600,miR-149,and PTK7 in both thyroid cancer tissues and thyroid cell lines.Thyroid cancer cell lines were cultured in vitro,and knockdown experiments were conducted for hsa_circ_0016600 and overexpression experiments for miR-149.Cell proliferation,migration,and invasion capabilities were assessed using CCK-8 assays,transwell assays,and scratch assays.A xenograft subcutaneous tumor model in mice was established to observe the tumorigenic effect of thyroid cancer cells with hsa_circ_0016600 knockdown in vivo.Fluorescence in situ hybridization experiments were conducted to assess the expression level of hsa_circ_0016600 in the xenograft tumor model in nude mice.RNA fluorescence in situ hybridization experiments were performed to determine the cellular localization of hsa_circ_0016600 and miR-149 in thyroid cancer cells.Dual-luciferase reporter gene assays were used to investigate the targeted regulatory relationships between hsa_circ_0016600 and miR-149,as well as miR-149 and PTK7.Results:In human thyroid cancer tissues and cell lines,the expression of hsa_circ_0016600(P<0.05)and PTK7(P<0.01)was upregulated,while the expression of miR-149(P<0.05)downregulated.Knockdown of hsa_circ_0016600 expression inhibited the proliferation(P<0.01),invasion(P<0.01),and migration(P<0.01)capabilities of thyroid cancer cells.Hsa_circ_0016600 could promote the proliferation(P<0.01),migration(P<0.01),and angiogenesis of thyroid cancer cells by targeting miR-149 and upregulating the expression of PTK7(P<0.01).Additionally,PTK7 was a target gene of miR-149.Conclusion:Hsa_circ_0016600 is highly expressed in thyroid cancer cells and promotes thyroid cancer cell migration by targeting regulation of the miR-149/PTK7 axis.