沉默LASP1对口腔鳞癌细胞生物学行为及3种抗肿瘤药物IC50的影响

doi:10.19439/j.sjos.2019.01.003

上海口腔医学 ›› 2019, Vol. 28 ›› Issue (1): 13-19.doi: 10.19439/j.sjos.2019.01.003

沉默LASP1对口腔鳞癌细胞生物学行为及3种抗肿瘤药物IC₅₀的影响

陈祺¹, 王旭², 余优成³, 魏魁杰¹

1.复旦大学附属中山医院青浦分院口腔科,上海 201700;
2.上海交通大学附属第九人民医院·口腔医学院口腔颌面-头颈肿瘤科,上海市口腔医学研究所,上海市口腔医学重点实验室,上海 200011;
3.复旦大学附属中山医院口腔科,上海 200032

收稿日期:2018-06-20 修回日期:2018-09-07 出版日期:2019-02-25 发布日期:2019-04-12
通讯作者: 魏魁杰,E-mail:wkj100310@163.com
作者简介:陈祺（1990-）,男,在读硕士研究生,住院医师,E-mail：cqbbm2008@126.com
基金资助:
上海市科学技术委员会科研计划项目（124119b0400）; 上海市卫生局科研课题计划（20134193）; 上海市青浦区科技发展基金医疗卫生项目(QKY2017-02)

Effect of silencing LASP1 on biological behaviors of oral squamous cell carcinoma cells and IC₅₀ of three anti-tumor drugs

CHEN Qi¹, WANG Xu², YU You-cheng³, WEI Kui-jie¹

1.Department of Stomatology, Qingpu Branch, Zhongshan Hospital, Fudan University. Shanghai 201700;
2. Department of Oromaxillofacial Head and Neck Oncology, Shanghai Ninth People's Hospital, College of Stomatology, Shanghai Jiao Tong University, Shanghai Research Institute of Stomatology, Shanghai Key Laboratory of Stomatology. Shanghai 200011;
3. Department of Stomatology, Zhongshan Hospital, Fudan University. Shanghai 200032, China

Received:2018-06-20 Revised:2018-09-07 Online:2019-02-25 Published:2019-04-12

摘要/Abstract

摘要： 目的:评价LASP1 对口腔鳞癌细胞增殖、转移、侵袭和周期的影响,并对3种抗肿瘤药物顺铂、阿帕替尼和多西他赛的相关作用进行分析。方法:利用The human protein atlas数据分析LASP1与头颈部肿瘤生存率、预后的关系。RT-PCR 和Western免疫印迹检测LASP1在口腔鳞癌细胞系的mRNA和蛋白表达。慢病毒构建LASP1沉默的HN30稳转细胞株,CCK-8 法检测细胞增殖,平板克隆实验检测细胞克隆形成能力,Transwell法检测细胞转移和侵袭能力,流式细胞仪检测细胞周期变化。建立裸鼠口腔鳞癌肺部转移瘤。CCK-8法分析3种药物顺铂、阿帕替尼和多西他赛在细胞中的IC₅₀变化。采用SPSS 11.0 软件包对数据进行统计学分析。结果:LASP1与头颈部肿瘤生存率、预后密切相关。LASP1促进口腔鳞癌细胞系HN30增殖、克隆形成、转移和侵袭,促进细胞周期G2/M期过渡。沉默LASP1后,裸鼠肺部转移瘤显著减少,多西他赛IC₅₀显著下调,而顺铂和阿帕替尼IC₅₀无显著变化。结论:LASP1促进口腔鳞癌细胞增殖、平板克隆、转移和侵袭,细胞周期G2/M期过渡,促进裸鼠体内肺转移瘤形成和多西他赛耐药。

关键词: 口腔鳞癌, LASP1, 细胞转移和侵袭, 转移瘤, 多西他赛

Abstract: PURPOSE: This study was designed to investigate the effects of LASP1 on proliferation, metastasis, invasion, and cycle of oral squamous cell carcinoma cells and analyze the changes of IC50 in three antitumor drugs: cisplatin, apatinib and docetaxel. METHODS: The correlation between LASP1 and survival rate and prognosis of patients with head and neck cancer were analyzed on the human protein atlas data. RT-PCR and Western blot were used to detect mRNA and protein expression of LASP1 in oral squamous cell carcinoma cell lines. LASP1 silenced HN30 stable transfectant cell line was constructed by lentivirus. CCK-8 assay was used to detect cell proliferation. Plate colony assay was used to detect cell clone formation ability. Transwell assay was used to detect cell migration and invasion ability. Flow cytometry was used to detect cell cycle changes. Oral squamous cell carcinoma metastases were established in nude mouse, the number of metastatic lung nodules was counted and stained with H-E. CCK-8 method was used to analyze the changes of IC50 in three antitumor drugs: cisplatin, apatinib and docetaxel. Statistical analysis was performed using SPSS 11.0 software package. RESULTS: LASP1 was closely related to the survival rate and prognosis of head and neck cancer. LASP1 promoted proliferation, colony formation, metastasis and invasion of oral squamous cell carcinoma cell line HN30, promoted G2/M phase transition of cell cycle, and significantly reduced the formation of lung metastasis in nude mice after silencing. There was significant correlation with docetaxel IC50 but no significant impact on cisplatin IC50 and aptatinib IC50. CONCLUSIONS: LASP1 enhances cell proliferation, plate cloning, metastasis and invasion, G2/M phase transition of cell cycle, promotes lung metastasis in nude mice and docetaxel resistance of oral squamous cell carcinoma cell line HN30.

Key words: Oral squamous cell carcinoma, LASP1, Cell metastasis and invasion, Metastatic lung nodules, Docetaxel

中图分类号:

R739.8

陈祺, 王旭, 余优成, 魏魁杰. 沉默LASP1对口腔鳞癌细胞生物学行为及3种抗肿瘤药物IC₅₀的影响[J]. 上海口腔医学, 2019, 28(1): 13-19.

CHEN Qi, WANG Xu, YU You-cheng, WEI Kui-jie. Effect of silencing LASP1 on biological behaviors of oral squamous cell carcinoma cells and IC₅₀ of three anti-tumor drugs[J]. Shanghai Journal of Stomatology, 2019, 28(1): 13-19.

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沉默LASP1对口腔鳞癌细胞生物学行为及3种抗肿瘤药物IC₅₀的影响

Effect of silencing LASP1 on biological behaviors of oral squamous cell carcinoma cells and IC₅₀ of three anti-tumor drugs

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