华西口腔医学杂志 ›› 2012, Vol. 30 ›› Issue (4): 350-355.doi: 10.3969/j.issn.1000-1182.2012.04.005

• 基础研究 • 上一篇    下一篇

微小核糖核酸-21对人舌鳞状细胞癌细胞系生物学特性的影响

王寅1,2 李龙江3 徐克4 吕品2 郑文龙5   

  1. 1.口腔疾病研究国家重点实验室, 四川大学, 成都610041;2.天津医科大学总医院口腔颌面外科, 天津300052; 3.四川大学华西口腔医院头颈肿瘤外科, 成都610041;4.天津医科大学总医院天津市肺癌研究所, 天津300052; 5.天津市疾病预防控制中心综合防治科, 天津300011
  • 收稿日期:2011-07-28 修回日期:2011-11-10 出版日期:2012-08-01 发布日期:2012-08-01
  • 通讯作者: 李龙江,Tel:028-85501428
  • 作者简介:王寅(1970—),女,天津人,副主任医师,博士
  • 基金资助:

    国家自然科学基金资助项目(30973345)

Biological effect of micro ribonucleic acid-21 on human tongue squamous cell carcinoma lines

Wang Yin1,2, Li Longjiang3, Xu Ke4, Lü Pin2, Zheng Wenlong5.   

  1. 1. State Key Laboratory of Oral Diseases, Sichuan University, Chengdu 610041, China; 2. Dept. of Oral and Maxillofacial Surgery, Tianjin Medical University General Hospital, Tianjin 300052, China; 3. Dept. of Head and Neck Oncology, West China School of Stomatology, Sichuan University, Chengdu 610041, China; 4. Tianjin Lung Cancer Institute, Tianjin Medical University General Hospital, Tianjin 300052, China; 5. Dept. of Comprehensive Prevention and Control, Tianjin Disease Prevention and Control Center, Tianjin 300011, China
  • Received:2011-07-28 Revised:2011-11-10 Online:2012-08-01 Published:2012-08-01

摘要:

目的研究微小核糖核酸-21(miR-21)对人舌鳞状细胞癌细胞系生物学特性的影响。方法将2’O-Me修饰的正义和反义miR-21寡核苷酸序列通过脂质体转染技术转染到舌鳞状细胞癌细胞Tca8113及其高转移株内,采用实时荧光定量聚合酶链式反应检测转染后miR-21在细胞中的表达变化,通过甲基噻唑基四唑(MTT)法、流式细胞术、Annexin Ⅴ细胞早期凋亡检测、划痕实验及Transwell实验分别检测细胞增殖能力、细胞周期、细胞早期凋亡情况、细胞迁移及侵袭能力的变化。结果靶向miR-21的反义寡核苷酸可以有效地抑制Tca8113及其高转移株细胞的增殖,促进细胞凋亡,并抑制细胞的迁移和侵袭能力。结论细胞内转染反义miR-21寡核苷酸序列后,可以降低舌鳞状细胞癌细胞中miR-21的表达,并对舌鳞状细胞癌细胞的生物学行为产生影响。

关键词: 舌鳞状细胞癌, 微小核糖核酸-21, 微小核糖核酸-21反义寡核苷酸, 生物学特性

Abstract:

Objective To detect the biological influence to human tongue squamous cell carcinoma(TSCC) cells of micro ribonucleic acid-21(miR-21). Methods Referring to mature miR-21 sequence, the sense and antisense oligonucleotide(sense-miR-21 and AS-miR-21) modified by 2’O-Me were designed to transfect into TSCC cells(Tca8113 and high metastasis cells) by liposome transfection technology, in order to establish an in vitro TSCC cell model. The expression changes of miR-21 in the transfected cells were detected with real-time fluorescence quantitative polymerase chain reaction(real-time PCR). The changes of cell proliferation, cell cycle, cell early apoptosis, cell migration and invasion capabilities were detected respectively by the technologies of methyl thiazolyl tetrazolium(MTT), flow cytometry,  Annexin Ⅴ cell early apoptosis assay, scratch assay and Transwell assay, to check AS-miR-21’s effect on the biological
characteristics of human TSCC cell lines. Results For the TSCC cells, the antisense oligonucleotide of targeting miR-21 could effectively inhibit cell proliferation, promoted cell apoptosis, and inhibited the capability of cell’s migration and invasion. Conclusion The expressions of miR-21 decrease after AS-miR-21 transfected into TSCC cells, and miR-21 can affect biological behavior of TSCC cells.

Key words: tongue squamous cell carcinoma, micro ribonucleic acid-21, antisense-micro ribonucleic acid-21 oligonucleotide, biological effect