唾液中微小RNA-375对口腔鳞状细胞癌的诊断价值及其对人口腔鳞状细胞癌细胞生物学功能的影响

doi:10.13701/j.cnki.kqyxyj.2019.09.016

口腔医学研究 ›› 2019, Vol. 35 ›› Issue (9): 883-888.DOI: 10.13701/j.cnki.kqyxyj.2019.09.016

唾液中微小RNA-375对口腔鳞状细胞癌的诊断价值及其对人口腔鳞状细胞癌细胞生物学功能的影响

谢龙¹, 桑磊^2,3*, 宋卓英⁴, 江燕军⁵

1. 武汉大学口腔医院湖北武汉 430079;
2. 苏州卫生职业技术学院江苏苏州 215000;
3. 苏州市华夏口腔医院口腔颌面外科江苏苏州 215000;
4. 辽宁省营口市中心医院口腔科辽宁营口 115003;
5. 中国煤炭总医院口腔科北京 100028

收稿日期:2019-01-25 出版日期:2019-09-28 发布日期:2019-09-25
通讯作者: 桑磊,E-mail:258470029@qq.com
作者简介:谢龙(1987~ ),男,湖北石首人,硕士,主治医师,主要从事口腔颌面外科临床研究工作。
基金资助:
苏州卫生职业技术学院院级课题(编号:SZWZY201315)

Diagnostic Values of Salivary MicroRNA-375 for OSCC and Its Effects on Biological Function of OSCC Cells

XIE Long¹, SANG Lei^2,3*, SONG Zhuoying⁴, JIANG Yanjun⁵

1. Hospital of Stomatology, Wuhan University, Wuhan 430079, China;
2. Suzhou Vocational Health College, Suzhou 215000, China;
3. Suzhou Huaxia Stomatological Hospital, Suzhou 215000, China;
4. Department of Stomatology, Yingkou Central Hospital, Yingkou 115003, China;
5. Department of Stomatology, China General Meitan Hospital, Beijing 100028, China.

Received:2019-01-25 Online:2019-09-28 Published:2019-09-25

摘要/Abstract

摘要： 目的:探讨口腔鳞状细胞癌(oral squamous cell carcinomas,OSCC)患者唾液中微小RNA(microRNA,miRNA,miR)-375对OSCC的诊断价值以及研究miR-375对人OSCC细胞株增殖、凋亡、迁移和侵袭等生物学功能的影响。方法:收集20例OSCC患者和20例健康成年人的唾液,采用实时定量聚合酶链反应(real-time quantitative polymerase chain reaction,Real-time PCR)检测唾液中miR-375的相对表达情况,miR-375 mimic/inhibitor转染细胞为实验组,未转染组细胞为对照组。Real-time PCR检测不同OSCC细胞系中miR-375的表达情况,脂质体转染法将miR-375 mimic/inhibitor分别转入SCC-4和HN-6细胞,Real-time PCR检测miR-375的表达情况;5-乙炔基-2'-脱氧尿嘧啶核苷(5-ethynyl-2’-deoxyuridine, EdU)掺入实验检测细胞增殖;流式细胞术检测miR-375对细胞凋亡的影响;划痕实验检测细胞的迁移和侵袭能力。结果:OSCC患者唾液中miR-375水平较健康成年人显著降低(P<0.05),在miR-375表达最弱的细胞株SCC-4中,转染miR-375 mimic后,抑制细胞活性,促进细胞凋亡,抑制细胞侵袭和迁移;而在miR-375表达最强的细胞株HN-6中,转染miR-375 inhibitor后, 促进细胞增殖,抑制细胞凋亡,促进细胞侵袭和迁移。结论:miR-375在 OSCC细胞中可起到抑癌基因的作用,与肿瘤的恶性程度有关,唾液miR-375将来有望作为OSCC诊断的标志物。

关键词: 微小RNA-375, 口腔鳞状细胞癌, 增殖, 凋亡, 迁移, 侵袭

Abstract: Objective: To explore the diagnostic value of salivary miR-375 for patients with oral squamous cell carcinomas (OSCC) and investigate the potential effects of miR-375 on biological characteristics of OSCC cells. Methods: Saliva samples from 20 OSCC patients and 20 healthy controls were collected. RT-PCR was applied to compare the relative expression of miR-375. SCC-4 cells were cultured and transfected with miR-375 mimics (miR-375 mimic group) via Lipofectamine 2000 liposome. HN-6 cells were cultured and transfected with miR-375 inhibitor (miR-375 inhibitor group) via Lipofectamine 2000 liposome. All cells without transfection were used as control group. The cell proliferation and cell apoptosis were measured by EDU and flow cytometry. The migration and invasion ability of the cells were detected by scratch test and Transwell test, respectively. Results: The expression of salivary miR-375 in OSCC patients was significantly lower than that in healthy control group (P<0.05). The low expression of miR-375 in SCC-4 cells, after miR-375 mimicking transfection, inhibited the cell proliferation, promoted the apoptosis, and decreased the cell migration and invasive ability. The highly expression of miR-375 in HN-6 cells, after miR-375 inhibitor transfection, promoted the cell proliferation, inhibited the apoptosis,and increased the cell migration and invasive ability. Conclusion: Our study indicates that miR-375 is down expression in the saliva of patients with OSCC and likes a tumor suppressor gene in OSCC, and it could be used as an indicator of diagnosis of OSCC in future.

Key words: microRNA-375, oral squamous cell carcinoma, proliferation, apoptosis, migration, invasion

谢龙, 桑磊, 宋卓英, 江燕军. 唾液中微小RNA-375对口腔鳞状细胞癌的诊断价值及其对人口腔鳞状细胞癌细胞生物学功能的影响[J]. 口腔医学研究, 2019, 35(9): 883-888.

XIE Long, SANG Lei, SONG Zhuoying, JIANG Yanjun. Diagnostic Values of Salivary MicroRNA-375 for OSCC and Its Effects on Biological Function of OSCC Cells[J]. Journal of Oral Science Research, 2019, 35(9): 883-888.

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唾液中微小RNA-375对口腔鳞状细胞癌的诊断价值及其对人口腔鳞状细胞癌细胞生物学功能的影响

Diagnostic Values of Salivary MicroRNA-375 for OSCC and Its Effects on Biological Function of OSCC Cells

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