论文部分内容阅读
目的构建人Snail基因慢病毒干扰载体及观察干扰前后对鼻咽癌5-8F细胞增殖和侵袭的影响。方法设计Snail基因特异性RNAi靶序列,克隆至经双酶切后的plVTHM线性化载体,测序鉴定正确后,用病毒上清感染鼻咽癌细胞5-8F,流式细胞仪分选获得稳定干扰Snail基因的细胞亚系。荧光定量PCR检测mRNA表达情况;MTT法和体外侵袭实验分别测定对细胞增殖和侵袭的影响。结果plVTHM-siSnail慢病毒干扰载体构建正确。荧光定量PCR结果表明分选后的细胞Snail mRNA水平表达显著降低。干扰后的细胞增殖减慢,穿透基膜的细胞数量减少,差异具有统计学意义(P<0.05)。结论plVTHM-siSnail表达质粒可显著下调Snail基因在5-8F中的表达,在一定程度上抑制鼻咽癌细胞的增殖和侵袭。
Objective To construct a lentiviral vector containing human Snail gene and study the effect of interfering on proliferation and invasion of nasopharyngeal carcinoma 5-8F cells. Methods Snail gene-specific RNAi target sequence was designed and cloned into plVTHM linearized vector after double digestion. After sequencing and identification, the nasopharyngeal carcinoma cells 5-8F were infected with virus supernatant, and stable interference was obtained by flow cytometry Snail gene cell subline. Fluorescent quantitative PCR was used to detect the mRNA expression. MTT assay and in vitro invasion assay were used to determine the effect on cell proliferation and invasion respectively. Results The plVTHM-siSnail lentiviral vector was constructed correctly. Fluorescent quantitative PCR results showed that the expression of Snail mRNA in the sorted cells was significantly reduced. After interference, the proliferation of cells slowed down and the number of cells penetrating the basement membrane decreased. The difference was statistically significant (P <0.05). Conclusion The plVTHM-siSnail expression plasmid can significantly down-regulate the expression of Snail gene in 5-8F and inhibit the proliferation and invasion of nasopharyngeal carcinoma cells to a certain extent.