目的 :观察小干扰RNA(small interference RNA,si RNA)沉默Gankyrin基因对喉鳞癌Hep-2细胞的生物学行为影响。方法:用si RNA沉默喉鳞癌Hep-2细胞中的Gankyrin基因,运用实时定量反转录聚合酶链反应(q RT-PCR)及蛋白印迹法(Western blot)技术检测转染前后Gankyrin m RNA及蛋白的表达。采用CCK-8法检测细胞增殖能力,流式细胞仪检测细胞凋亡及细胞周期,细胞划痕损伤实验和Transwell小室实验检测细胞迁移能力,Matrigel侵袭实验检测细胞侵袭能力,Western blot检测Gankyrin下调后p53蛋白的表达。结果:q RT-PCR和Western blot结果显示,Gankyrin si RNA组Gankyrin m RNA和蛋白的相对表达量均低于对照si RNA组和未处理组(P<0.001)。CCK-8法显示Gankyrin si RNA组细胞增殖速度明显下降(P<0.001)。流式细胞仪检测结果表明,Gankyrin si RNA组细胞凋亡率[(7.70±1.12)%]明显高于对照si RNA组[(2.34±0.32)%]和未处理组[(1.82±0.29)%],差异有统计学意义(P<0.001);与两对照组相比,Gankyrin si RNA组G1期比例增加,S期比例减少,差异有统计学意义(P<0.001)。细胞划痕损伤实验和Transwell小室实验显示Gankyrin si RNA组的细胞迁移能力明显减弱(P<0.01);Matrigel侵袭实验显示Gankyrin si RNA组细胞侵袭能力与对照si RNA组和未处理组比较,差异无统计学意义(P>0.05)。Gankyrin表达下调增加了Hep-2细胞中p53蛋白的表达,差异有统计学意义(P<0.001)。结论:Gankyrin表达下调能抑制Hep-2细胞的增殖和迁移,可能与细胞凋亡、细胞周期改变及p53的表达密切相关。 Objective: To observe the effect of Gankyrin gene silencing small interfering RNA (siRNA) on the biological behavior of Hep-2 cells. Methods: The Gankyrin gene was silenced by si RNA in Hep-2 cells. The expression of Gankyrin m RNA was detected by q RT-PCR and Western blot, And protein expression. Cell proliferation was detected by CCK-8 assay. Cell apoptosis and cell cycle were detected by flow cytometry. Scratch cell injury assay and Transwell chamber assay were used to detect cell migration. Matrigel invasion assay was used to detect cell invasion. Western blot was used to detect Gankyrin downregulation p53 protein expression. Results: q-RT-PCR and Western blot showed that the relative expression levels of Gankyrin m RNA and protein in Gankyrin si RNA group were lower than those in control si RNA group and untreated group (P <0.001). The CCK-8 assay showed that the proliferation rate of Gankyrin si RNA group was significantly decreased (P <0.001). The results of flow cytometry showed that the apoptosis rate in Gankyrin si RNA group was significantly higher than that in control si RNA group [(2.34 ± 0.32)%] and (1.82 ± 0.29)% in untreated group [(7.70 ± 1.12)%] ], The difference was statistically significant (P <0.001). Compared with the two control groups, Gankyrin si RNA increased the proportion of G1 phase, S phase decreased, the difference was statistically significant (P <0.001). The cell scratch injury assay and Transwell chamber assay showed that the cell migration ability of Gankyrin si RNA group was significantly decreased (P <0.01). Matrigel invasion assay showed that the cell invasion ability of Gankyrin si RNA group was significantly lower than that of the control si RNA group and the untreated group Statistical significance (P> 0.05). The downregulation of Gankyrin increased the expression of p53 protein in Hep-2 cells, the difference was statistically significant (P <0.001). Conclusion: Down-regulation of Gankyrin can inhibit the proliferation and migration of Hep-2 cells, which may be closely related to apoptosis, cell cycle changes and p53 expression.