摘要
目的研究针对自分泌运动因子(autotaxin,ATX)的小干扰RNA(siRNA)对人胆管癌细胞ATX基因表达的影响,探讨RNA干扰技术治疗胆管癌的前景。方法参考人黑色素瘤ENPP2基因序列,设计合成ATX-siRNA及随机阴性对照。在阳离子脂质体的介导下转染人胆管癌细胞HCCC-9810。分别于转染后24,48,72h收集细胞,用半定量RT-PCR方法检测转染后ATXmRNA表达的变化,并与对照组及ATX表达抑制剂IL-1β的作用进行比较。结果体外合成的siRNA在转染胆管癌细胞24h后ATXmRNA的表达降低,48h抑制作用最为明显,与对照组比较差异有显著性意义(P<0.01),并明显强于IL-1β(P<0.01)。结论siRNA可有效地抑制胆管癌细胞ATX的表达,从而有可能阻断ATX对肿瘤细胞运动的促进作用。
Objective To investigate the effects of siRNA on expression of ATX mRNA in human biliary cancer cell HCCC-9810.Methods According to ATX gene sequence, an ATX-siRNA and a nagtive siRNA were designed and synthysized. The siRNAs were transfected into biliary cancer cell line HCCC-9810 with the media of Lipofectamine 2000. At 24, 48 and 72 h post-transfection, the cells were collected and analyzed. The changes of ATX mRNA expressions were detected by semi-quantitative RT-PCR. Results The ATX siRNA specifically targeting ATX mRNA was successfully constructed. Semi-quantitive RT-PCR assay showed that a decrease of ATX mRNA expression was detected at 24 h post-transfection, with maximal inhibition at 48 h post-transfection, but no further reduction at 72 h post-transfection. The ATX mRNA expression treated with siRNA for 48 h were decreasted signficantlly compared to that of controls and that treated with IL-1β （ P 〈 0.01 ）. Neither of Lipofectamine or siRNA-co groups showed marked reduction of ATX mRNA expression. Conclusion The ATXsiRNA can reduce the expression of ATX mRNA level, which is morn efficient than that of IL-1β. Therefore, ATX siRNA may be used for further study to inhibit tumor spread in vivo experiments.
出处
《肝胆胰外科杂志》
CAS
2006年第5期291-293,共3页
Journal of Hepatopancreatobiliary Surgery
基金
江苏省教委自然科学基金资助项目(02KJD320033)
