[1]周美岑,兰玲,邓微,等.MCPIP4诱导甲状腺乳头状癌细胞系TPC-1周期停滞的机制[J].国际内分泌代谢杂志,2023,43(02):86-90,95.[doi:10.3760/cma.j.cn121383-20210605-06008]
　Zhou Meicen,Lan Ling,Deng Wei,et al.The mechanism of RNA binding protein MCPIP4 induces cell cycle arrest in papillary thyroid cancer[J].International Journal of Endocrinology and Metabolism,2023,43(02):86-90,95.[doi:10.3760/cma.j.cn121383-20210605-06008]
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MCPIP4诱导甲状腺乳头状癌细胞系TPC-1周期停滞的机制()
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《国际内分泌代谢杂志》[ISSN:1673-4157/CN:12-1383/R]

卷:
43
期数:
2023年02期
页码:
86-90,95
栏目:
论著
出版日期:
2023-03-20

文章信息/Info

Title:
The mechanism of RNA binding protein MCPIP4 induces cell cycle arrest in papillary thyroid cancer
作者:
周美岑1; 兰玲1; 邓微1; 鹿文葆2
1北京积水潭医院 北京大学第四临床医学院内分泌科,北京 100035; 2中国医学科学院&北京协和医学院 微循环研究所,北京 100005
Author(s):
Zhou Meicen1;  Lan Ling1;  Deng Wei1;  Lu Wenbao2.
1Department of Endocrinology, Beijing Jishuitan Hospital, the Fourth Clinical College of Peking University, Beijing 100035, China; 2Institute of Microcirculation, Chinese Academy of Medical Sciences(CAMS)& Peking Union Medical School(PUMC), Beijing 100005, China
关键词:
甲状腺乳头状癌;  单核细胞趋化蛋白诱导蛋白4;  TPC-1;  细胞周期
Keywords:
Papillary thyroid carcinoma;  Monocyte chemotactic protein-induced protein 4;  TPC-1;  Cell cycle
DOI:
10.3760/cma.j.cn121383-20210605-06008
摘要:
目的 探讨RNA结合蛋白——单核细胞趋化蛋白诱导蛋白4(MCPIP4)在甲状腺乳头状癌细胞中的作用及机制。方法 用脂质体转染法分别在甲状腺乳头状癌细胞系TPC-1细胞中过表达外源性MCPIP4或靶向人MCPIP4基因的shRNA,并筛选稳定表达单细胞克隆; 四甲基偶氮唑蓝(MTT)法检测细胞增殖; 流式细胞计量技术检测细胞周期; 实时荧光定量聚合酶链反应(qPCR)检测靶基因半衰期; RNA免疫沉淀法(RNA-IP)检测MCPIP4结合的靶基因mRNAs; 荧光素酶报告基因实验检测靶向基因3'非编码区的能力。结果 甲状腺乳头状癌细胞系TPC-1中MCPIP4 mRNA表达水平显著低于正常甲状腺组织; 过表达MCPIP4显著抑制TPC-1细胞增殖(P<0.05),而敲低MCPIP4显著促进该细胞增殖(P<0.05); 并分别显著增加或降低G1期细胞百分比(P<0.05); MCPIP4显著降低细胞周期素依赖性激酶(CDK)4、CDK6、周期蛋白D1和周期蛋白E1mRNAs的半衰期(P均<0.01); MCPIP4结合上述细胞周期相关基因(P<0.05); MCPIP4显著降低含不同3'UTR的报告基因荧光素酶活性(P<0.05)。结论 MCPIP4在甲状腺乳头状癌细胞TPC-1中发挥抑癌作用,诱导细胞周期G1期停滞可能是其发挥抑癌功能的机制之一。
Abstract:
Objective To investigate the functions of monocyte chemotactic protein-induced protein 4(MCPIP4)in human papillary thyroid cancer cell line TPC-1.Methods TPC-1 cells were transfected with GFP-tagged MCPIP4 by Tet-on inducing expression system. Endogenous MCPIP4 was knocked down by stable expressing shRNA. MTT assay was performed to measure the growth of TPC-1 cells after overexpression or knockdown of MCPIP4. FACS method was used to analysis cell cycle in TPC-1 cells. Real-time PCR was used to test the expression of cell cycle-related mRNAs expression and their half-life. RNA-IP experiment was conducted to detect the mRNA directly enriched by MCPIP4. Luciferase assay was performed to determine whether the mRNA decay was mediated through 3'UTR.Results The expression of MCPIP4 mRNA in papillary thyroid carcinoma cell line TPC-1 was significantly lower than that in normal thyroid tissue; MCPIP4 overexpression inhibited cell proliferation significantly(P<0.05), while knockdown MCPIP4 promoted cell proliferation with statistical significance(P<0.05). MCPIP4 induced cell cycle arrest in TPC-1 with statistical significance(P<0.05). MCPIP4 overexpression reduced the half-life of cell cycle mRNAs(CDK4, CDK6, Cyclin D1, Cyclin E1, respectively)with significance(all P<0.01). In addition, cell cycle-related mRNAs were able to be pulled down by GFP-MCPIP4 but not by isotype IgG(P<0.05). Compared with control vector, MCPIP4 significantly suppressed luciferase activities of all four 3'UTR reporters(P<0.05).Conclusion MCPIP4 can function as a tumor suppressor in human papillary thyroid cancer cell line TPC-1 through inducing G1 cell cycle arrest.

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备注/Memo

备注/Memo:
基金项目:北京积水潭医院院内青年基金(QN-201921); 北京积水潭医院自然基金培育计划(ZR-202115); 北京积水潭医院学科骨干人才培训计划(XKGG202119); 国家自然科学基金面上项目(81372858)
通信作者:鹿文葆,Email: luwenbao_217@163.com
更新日期/Last Update: 2023-04-15