论文部分内容阅读
目的:观察Hpa小干扰RNA(small-interfering RNA,siRNA)联合亚砷酸对胰腺癌侵袭力的影响,为Hpa-siRNA联合亚砷酸治疗胰腺癌提供实验依据。方法:将3条Hpa干扰靶序列Hpa1-siRNA,Hpa2-siRNA,Hpa3-siRNA稀释退火,分别与线性化pGenesil-1.1质粒表达载体的连接制成pGenesil1.1/Hpa1-siRNA、pGen-esil1.1/Hpa2-siRNA、pGenesil1.1/Hpa3-siRNA等重组质粒。然后将上述目的基因转染SW1990胰腺癌细胞,并设空白对照和阴性对照组。RT-PCR检测转染后Hpa-mRNA表达,Western blot检测转染后Hpa蛋白表达。采用Transwell小室试验检测转染后Hpa-siRNA和经亚砷酸处理SW1990细胞体外侵袭能力。结果:Hpa-mR-NA和Hpa蛋白在基因转染前后的表达,NC和HK组无明显差别,Hpa1-siRNA,Hpa2-siRNA,Hpa3-siRNA三组均明显下降,Hpa2-siRNA抑制效率优于Hpa1-siRNA和Hpa3-siRNA(P<0.05)。随着亚砷酸浓度从0.5、1.0、2.0mg/ml递增,侵袭抑制率也随之升高,分别为7.78%、46.8%及76.8%,差异有统计学意义(χ2=71.633,P<0.05)。Hpa2-siRNA联合亚砷酸后均能显著抑制SW1990胰腺癌细胞侵袭力,抑制率均达100%。结论:Hpa-siRNA和亚砷酸对SW1990胰腺癌细胞侵袭力具有协同抑制作用。
OBJECTIVE: To observe the effect of small interfering RNA (siRNA) combined with arsenious acid (Hpa) on the invasiveness of pancreatic cancer and provide the experimental evidence for the treatment of pancreatic cancer with Hpa-siRNA combined with arsenious acid. METHODS: Three Hpa interfering target sequences Hpa1-siRNA, Hpa2-siRNA and Hpa3-siRNA were diluted and annealed, respectively, and ligated with the linearized pGenesil-1.1 plasmid expression vector to prepare pGenesil1.1 / Hpa1-siRNA and pGen-esil1.1 / Hpa2-siRNA, pGenesil1.1 / Hpa3-siRNA and other recombinant plasmids. The target gene was then transfected into SW1990 pancreatic cancer cells, and blank control and negative control groups were established. Hpa-mRNA expression was detected by RT-PCR and Hpa protein expression by Western blot. Transwell chamber assay was used to detect the in vitro invasion ability of Hpa-siRNA and SW-9090 cells treated with arsenite. Results: There was no significant difference between Hpa1-siRNA, Hpa2-siRNA and Hpa3-siRNA in the expression of Hpa-mR-NA and Hpa before and after gene transfection. The inhibitory efficiency of Hpa2-siRNA was better than that of Hpa1-siRNA Hpa1-siRNA and Hpa3-siRNA (P <0.05). As the concentration of arsenious acid increased from 0.5, 1.0, 2.0mg / ml, the invasion inhibition rate also increased, which was 7.78%, 46.8% and 76.8% respectively, with significant difference (χ2 = 71.633, P <0.05 ). Hpa2-siRNA combined with arsenious acid could significantly inhibit the invasiveness of SW1990 pancreatic cancer cells, the inhibition rate reached 100%. Conclusion: Hpa-siRNA and arsenious acid have a synergistic inhibitory effect on invasiveness of SW1990 pancreatic cancer cells.