Wang Yin, Li Longjiang, Xu Ke, Lü Pin, Zheng Wenlong
State Key Laboratory of Oral Diseases, Sichuan University, Chengdu 610041, China.
Hua Xi Kou Qiang Yi Xue Za Zhi. 2012 Aug;30(4):350-5, 359.
To detect the biological influence to human tongue squamous cell carcinoma (TSCC) cells of micro ribonucleic acid-21 (miR-21).
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.
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.
The expressions of miR-21 decrease after AS-miR-21 transfected into TSCC cells, and miR-21 can affect biological behavior of TSCC cells.
检测微小核糖核酸-21(miR-21)对人舌鳞状细胞癌(TSCC)细胞的生物学影响。
参照成熟miR-21序列,设计2'-O-甲基修饰的正义和反义寡核苷酸(正义-miR-21和反义-miR-21),采用脂质体转染技术转染TSCC细胞(Tca8113和高转移细胞),建立体外TSCC细胞模型。用实时荧光定量聚合酶链反应(实时PCR)检测转染细胞中miR-21的表达变化。分别采用甲基噻唑基四氮唑(MTT)法、流式细胞术、膜联蛋白V细胞早期凋亡检测法、划痕试验和Transwell试验检测细胞增殖、细胞周期、细胞早期凋亡、细胞迁移和侵袭能力的变化,以观察反义-miR-21对人TSCC细胞系生物学特性的影响。
对于TSCC细胞,靶向miR-21的反义寡核苷酸可有效抑制细胞增殖,促进细胞凋亡,并抑制细胞迁移和侵袭能力。
反义-miR-21转染TSCC细胞后miR-21表达降低,且miR-21可影响TSCC细胞的生物学行为。