摘要
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 invasion inhibiting effect of Hpa small - interfering RNA （siRNA） combined with arsenous acid on pancreatic cancer cells, so as to provide the theoretical foundation for further clinical use in the cancer treatment. Methods ： Target gene （ Hpal - siRNA, Hpa2 - siRNA, Hpa3 - siRNA） fragments were diluted, annealed and connected with linear plasmid expression vector pGenesil - 1.1, then pGenesill. 1/Hpal - siRNA, pGenesill. 1/Hpa2 - siRNA and pGenesill. 1/Hpa3 - siRNA were reconstructed. The Hpa - siRNA was transfected into SW1990 pancreatic cancer cell by DOTAP Liposomes meanwhile the empty control and negative control group were designed. Those target genes were respectively transfected into Hpa - mRNA and Hpa protein expression was detected by RT - PCR and Western blot respectively. Transwell cab model was employed to test the ability of SW1990 cell. Results ： Expression of Hpa - mR- NA and Hpa - protein in SW1990 were obviously suppressed by Hpal - siRNA, Hpa2 - siRNA, and Hpa3 - siRNA, and efficacy in Hpa2 - siRNA better than in Hpal - siRNA and Hpa3 - siRNA （ P 〈 0.05 ）. With the improvement of arsenous acid concentration from 0.5, 1.0, to 2.0mg/mL, the invasive inhibitory rates in pancreatic cancer increased from 7. 78% , 46.8%, to 76.8%, respectively, the differences had statistics significance （ x^2 = 71. 633, P 〈 0. 001 ）. Inva- sive ability of pancreatic cancer cells were completely inhabited by Hpa2 - siRNA combined with arsenous acid （ invasive inhibitory rates reached 100% ）. Conclusion： Hpa- siRNA and arsenous acid have co-inhibition effect on pancreatic cancer invasion.
出处
《中华中医药学刊》
CAS
2011年第12期2657-2661,I0021,共6页
Chinese Archives of Traditional Chinese Medicine
基金
浙江省研究生创新科研资助项目(YK2008071)
关键词
乙酰肝素酶
小片段干扰RNA
亚砷酸
胰腺癌
侵袭力
Hparinase
small interfering RNA
arsenous acid
pancreatic cancer
invasive ability
