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目的:通过RNAi方法沉默SP1(转录因子特异性蛋白1)基因研究其对口腔鳞癌细胞KB的生物学功能的影响。方法:构建靶向人SP1基因的干扰序列,采用脂质体介导的转染技术沉默SP1基因的表达,同时采用CCK8法观察其对细胞增殖活性的影响,采用流式细胞仪测定对细胞周期分布的影响,采用Western blotting技术检测细胞凋亡相关蛋白的变化。结果:SP1沉默后能够有效抑制口腔鳞癌细胞KB的增殖活力,24、48、72、96 h抑制率分别为28.58%±3.42%、40.62%±5.78%、65.5%±5.27%、和79.63%±8.23%,与对照组相比,差异有统计学意义(P<0.05)。细胞周期实验显示,下调SP1表达可抑制口腔鳞癌细胞KB阻滞于G1期。SP1细胞增殖被明显抑制,Western blot检测发现,沉默SP1后Bcl-2表达下调,bax,bik表达上升。结论:SP1可能参与调控口腔鳞癌细胞KB的增殖凋亡,通过促进早期的凋亡以及G1期细胞周期阻滞而发挥抗口腔肿瘤增殖的能力,为以后临床开展SP1为靶点的口腔鳞癌基因治疗提供新的研究思路。
OBJECTIVE: To investigate the effect of silencing SP1 (transcription factor-specific protein 1) gene on the biological function of oral squamous cell carcinoma KB cells by RNAi. Methods: The targeting sequence of human SP1 gene was constructed. The expression of SP1 gene was silenced by liposome-mediated transfection. At the same time, the effect of SP1 gene on cell proliferation was observed by CCK8 assay. The cell cycle was determined by flow cytometry Distribution of the impact of the use of Western blotting detection of apoptosis-related protein changes. Results: Silencing SP1 could effectively inhibit the proliferation of oral squamous cell carcinoma KB cells, the inhibitory rates were 28.58% ± 3.42%, 40.62% ± 5.78%, 65.5% ± 5.27%, 79.63% at 24, 48, ± 8.23%, compared with the control group, the difference was statistically significant (P <0.05). Cell cycle experiments show that down-regulation of SP1 expression can inhibit KB cells in oral squamous cell carcinoma in G1 phase. SP1 cell proliferation was significantly inhibited, Western blot showed that after silencing SP1 Bcl-2 expression was down, bax, bik expression increased. Conclusion: SP1 may be involved in the regulation of proliferation and apoptosis of oral squamous cell carcinoma KB cells, which can promote the proliferation of oral cancer cells by promoting early apoptosis and G1 cell cycle arrest. Gene therapy provides new research ideas.