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敲低 KRAB 结构域相关蛋白 1 通过调控 P68/DEAD 盒蛋白 5 抑制甲状腺癌细胞的增殖、迁移和侵袭。

Knockdown of KRAB domain-associated protein 1 suppresses the proliferation, migration and invasion of thyroid cancer cells by regulating P68/DEAD box protein 5.

机构信息

Department of Thyroid and Breast Surgery, Zhongnan Hospital of Wuhan University, Wuhan, Hubei, China.

出版信息

Bioengineered. 2022 May;13(5):11945-11957. doi: 10.1080/21655979.2022.2067289.

Abstract

KRAB domain-associated protein 1 (KAP-1) has been reported to be an oncogene in diverse tumors. KAP-1 was found to have abundant existence in malignant thyroid tissues, but its role in thyroid cancer hasn't been elucidated clearly. This study was carried out to explore the role of KAP-1 in thyroid cancer, and to clarify its molecular mechanism. The expressions of KAP-1 and P68/DEAD box protein 5 (DDX5) were assessed under the help of qRT-PCR and western blot. Then, we downregulated KAP-1 or upregulated DDX5 by cell transfection in TPC-1 cells. A series of cellular experiments on proliferation, apoptosis, migration and invasion were conducted with CCK-8, EdU, TUNEL, wound-healing and Transwell assays. Besides, the relationship between KAP-1 and DDX5 was verified by co-immunoprecipitation (Co-IP). The results showed that both of KAP-1 and DDX5 were upregulated in thyroid cancer cells. Loss-of-function experiments revealed that KAP-1 knockdown imparted suppressive effects on cell proliferation, migration and invasion, but promoted cell apoptosis. Additionally, KAP-1 was demonstrated to interact with DDX5 and positively regulate DDX5 expression. The following rescued experiments exhibited that the inhibitory effects of KAP-1 knockdown on cellular activities of thyroid cancer and Wnt/β-catenin signaling were also partly reversed by DDX5 overexpression. Moreover, activation of Wnt/β-catenin signaling retarded the anti-tumor activity of KAP-1 knockdown. In conclusion, the data in this study disclosed that KAP-1 silence helped to repress the cell proliferation, migration and invasion by degrading DDK5, so as to hinder the development of thyroid cancer.

摘要

KAP1 相关蛋白 1(KAP-1)已被报道在多种肿瘤中为致癌基因。研究发现 KAP-1 在恶性甲状腺组织中大量存在,但在甲状腺癌中的作用尚未阐明。本研究旨在探讨 KAP-1 在甲状腺癌中的作用,并阐明其分子机制。采用 qRT-PCR 和 Western blot 评估 KAP-1 和 P68/DEAD 盒蛋白 5(DDX5)的表达。然后,我们通过细胞转染在 TPC-1 细胞中下调 KAP-1 或上调 DDX5。用 CCK-8、EdU、TUNEL、划痕愈合和 Transwell 测定法进行一系列细胞增殖、凋亡、迁移和侵袭实验。此外,通过共免疫沉淀(Co-IP)验证 KAP-1 和 DDX5 之间的关系。结果表明,甲状腺癌细胞中 KAP-1 和 DDX5 均上调。功能丧失实验表明,KAP-1 敲低对细胞增殖、迁移和侵袭具有抑制作用,但促进细胞凋亡。此外,证明 KAP-1 与 DDX5 相互作用并正向调节 DDX5 表达。后续的挽救实验表明,DDX5 过表达部分逆转了 KAP-1 敲低对甲状腺癌细胞活力和 Wnt/β-catenin 信号的抑制作用。此外,Wnt/β-catenin 信号的激活削弱了 KAP-1 敲低的抗肿瘤活性。综上所述,本研究的数据揭示了 KAP-1 沉默通过降解 DDK5 有助于抑制细胞增殖、迁移和侵袭,从而阻碍甲状腺癌的发展。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/55f1/9275928/aad93b47a158/KBIE_A_2067289_UF0001_OC.jpg

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