小分子RNA干扰沉默Tiam1基因对舌鳞状细胞癌细胞生物学特性的影响

Effect of Silencing Tiam1 Gene by Small RNA Interference on Biological Characteristics of Tongue Squamous Cell Carcinoma Cells

目的:探讨小分子RNA干扰(siRNA)沉默Tiam1基因对舌鳞状细胞癌细胞生物学特性的影响。方法:培养人舌癌细胞SCC-4,随机分为siRNA-Tiam1组、siRNA-对照序列组和空白对照组,利用实时荧光定量PCR技术和Western blot法检测不同转染组细胞中Tiam1基因和蛋白表达,MTT法检测不同转染组细胞增殖情况,利用流式细胞术检测不同转染组细胞凋亡情况,Transwell法检测不同转染组细胞迁移和侵袭能力。结果:siRNATiam1组细胞中Tiam1基因和蛋白相对表达量均低于siRNA-对照序列组和空白对照组(P<0.05);MTT实验结果显示,与siRNA-对照序列组和空白对照组相比,siRNA-Tiam1组细胞24h、48h、72h和96h时A值均降低(P<0.05);流式细胞检测结果显示,siRNA-Tiam1组细胞凋亡率显著高于siRNA-对照序列组和空白对照组(P<0.05);siRNA-Tiam1组迁移细胞数和侵袭细胞数均低于siRNA-对照序列组和空白对照组(P<0.05)。结论:特异性沉默Tiam1基因可有效抑制舌鳞状细胞癌细胞增殖能力,加速细胞凋亡,抑制细胞迁移和侵袭能力。

Objective： To investigate the effect of silencing Tiam1 gene by small RNA interference （siRNA） on the biological characteristics of tongue squamous cell carcinoma cells. Methods： The human tongue cancer cells of SCC-4 were cultured. All cells were randomly divided into siRNA-Tiam1 group, siRNA-control sequence group, and blank control group. The expressions of Tiam1 gene and protein in different transfection groups were detected by real-time fluorescence quantitative PCR and Western blot, respectively. The cell proliferations in different transfection groups were detected by MTT assay. The cell apoptosis in different transfection groups were detected by flow cytometry. The cell migration and invasion in different transfection groups were detected by Transwell assay. Results： The relative expression levels of Tiam1 gene and protein in the siRNA-Tiam1 group were lower than those in the siRNA-control sequence group and blank control group （P〈0.05）. MTT assay showed that, compared with siRNA-control sequence group and blank control group, the absorbance A values at 24h, 48h, 72h and 96h in the siRNA-Tiam1 group decreased （P〈0.05）. Flow cytometry showed that the apoptosis rate in the siRNA-Tiam1 group was significantly higher than that in the siRNA-control sequence group and blank control group （P〈0.05）. The number of migrating cells and invasive cells in the siRNA-Tiam1 group was lower than that in the siRNA-control sequence group and blank control group （P〈0.05）. Conclusion： Specific silencing Tiam1 gene could effectively inhibit the proliferation of tongue squamous cell carcinoma cells, accelerate cell apoptosis, and inhibit cell migration and invasion ability.