POLD1对口腔鳞癌细胞增殖、侵袭的影响及调控机制

doi:10.19438/j.cjoms.2021.04.003

中国口腔颌面外科杂志 ›› 2021, Vol. 19 ›› Issue (4): 302-308.doi: 10.19438/j.cjoms.2021.04.003

POLD1对口腔鳞癌细胞增殖、侵袭的影响及调控机制

张莹^1,2, 吴利², 陈羽菱², 林云涛², 沈月洪^2,*, 杨宏宇^1,2,*

1.北京大学口腔医学院·口腔医院综合科,国家口腔疾病临床医学研究中心, 口腔数字化医疗技术和材料国家工程实验室,口腔数字医学北京重点实验室,北京 100081;
2.北京大学深圳医院口腔医学中心口腔颌面外科,广东省高水平临床重点专科, 广东省口腔疾病诊疗技术工程技术研究中心,广东深圳 518036

收稿日期:2021-01-27 修回日期:2021-03-22 出版日期:2021-07-20 发布日期:2021-08-05
通讯作者: 沈月洪, E-mail：yuehongshen@hotmail.com;杨宏宇, E-mail：hyyang192@hotmail.com。^*共同通信作者
作者简介:张莹（1993-）,女,硕士研究生,E-mail:1811210573@bjmu.edu.cn
基金资助:
广东省基础与应用基础研究基金（2019A1515011911）; 深圳市科技创新基础研究基金（JCYJ20200109140208058,JCYJ20190809104803572）; 广东省高水平临床重点专科（深圳市配套建设经费,SZGSP008）

Effect of POLD1 on proliferation and invasion of oral squamous cell carcinoma and its regulatory mechanism

ZHANG Ying^1,2, WU Li², CHEN Yu-ling², LIN Yun-tao², SHEN Yue-hong², YANG Hong-yu^1,2

1. Department of General Dentistry, Peking University School and Hospital of Stomatology, National Clinical Research Center for Oral Diseases, National Engineering Laboratory of Dental Digital Medical Technology and Materials, Beijing Key Laboratory of Oral Digital Medicine.Beijing 100081;
2. Department of Oral and Maxillofacial Surgery, Stomatological Center, Peking University Shenzhen Hospital, Guangdong Provincial High-Level Clinical Key Specialty, Guangdong Province Engineering Research Center of Oral Disease Diagnosis and Treatment. Shenzhen 518036, Guangdong Province, China

Received:2021-01-27 Revised:2021-03-22 Online:2021-07-20 Published:2021-08-05

摘要/Abstract

摘要： 目的： 探讨人源DNA 聚合酶δ催化亚基（POLD1）的表达对口腔鳞状细胞癌（oral squamous cell carcinoma,OSCC）细胞增殖、迁移、侵袭、细胞周期的影响及其相关机制。方法： 使用生物信息数据分析POLD1在头颈部鳞癌及癌旁组织中的表达,利用免疫组织化学染色检测POLD1在40对口腔鳞癌及癌旁组织中的表达。慢病毒感染SCC9及CAL27细胞系,蛋白免疫印迹检测POLD1在稳转细胞系中的表达。CCK-8实验和EdU实验检测细胞增殖能力,划痕实验和Transwell迁移和侵袭实验检测细胞的迁移和侵袭能力。流式细胞周期实验检测POLD1对细胞周期的影响。蛋白免疫印迹法研究POLD1影响细胞周期的相关通路。采用SPSS 21.0软件包和GraphPad Prism 进行数据分析和绘图。结果： POLD1在头颈部鳞癌及40对OSCC组织中的表达显著高于癌旁组织（P<0.001);慢病毒感染SCC9及CAL27细胞系后,POLD1的表达显著下降,且在细胞核及细胞质中的表达降低。实验组SCC9及CAL27细胞的增殖、迁移、侵袭能力下降。流式细胞周期实验检测到实验组细胞在G2/M期阻滞,G1期缩短。蛋白免疫印迹结果： 显示,实验组P-Cdc2(Tyr15)、P-Rb(Ser807)表达显著升高。结论： 敲低POLD1的表达可抑制OSCC细胞增殖、迁移和侵袭。

关键词: 口腔鳞癌, POLD1, 增殖, 迁移, 侵袭, 细胞周期/G2期阻滞, P-Cdc2

Abstract: PURPOSE: To investigate the effect of POLD1 expression on cell proliferation, migration, invasion and cell cycle of oral squamous cell carcinoma (OSCC) cell lines and the relevant mechanism. METHODS: Online data were used to analyze POLD1 expression in head and neck squamous cell carcinoma (HNSCC) samples and normal tissues. POLD1 expression was evaluated in OSCC and normal tissues using immunohistochemistry. OSCC cell lines（SCC9 and CAL27) were infected with lentivirus, and the expression of POLD1 in stable cell lines was detected by Western blot. CCK-8 assay and EdU assay were used to detect cell proliferation; wound healing assay and Transwell migration and invasion assay were used to detect cell migration and invasion. The effect of POLD1 on cell cycle was detected by flow cytometry and the related pathway was studied via Western blot. SPSS 21.0 software package and GraphPad Prism were used for data analysis and plotting. RESULTS: POLD1 was abnormally upregulated in head and neck squamous cell carcinoma and OSCC tissues (P<0.001). Depletion of POLD1 in OSCC cells not only inhibited proliferation, migration, and invasion of cancer cells but also resulted in G2 arrest. The expression of P-Cdc2(Tyr15) and P-Rb(Ser807) in the experiment group were significantly increased. CONCLUSIONS: Down-regulation of POLD1 inhibits proliferation, migration and invasion of OSCC cells.

Key words: Oral squamous cell carcinoma, DNA polymerase delta subunit 1, Proliferation, Migration, Invasion, Cell cycle/G2 arrest, P-Cdc2

中图分类号:

R739.8

张莹, 吴利, 陈羽菱, 林云涛, 沈月洪, 杨宏宇. POLD1对口腔鳞癌细胞增殖、侵袭的影响及调控机制[J]. 中国口腔颌面外科杂志, 2021, 19(4): 302-308.

ZHANG Ying, WU Li, CHEN Yu-ling, LIN Yun-tao, SHEN Yue-hong, YANG Hong-yu. Effect of POLD1 on proliferation and invasion of oral squamous cell carcinoma and its regulatory mechanism[J]. China J Oral Maxillofac Surg, 2021, 19(4): 302-308.

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POLD1对口腔鳞癌细胞增殖、侵袭的影响及调控机制

Effect of POLD1 on proliferation and invasion of oral squamous cell carcinoma and its regulatory mechanism

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