小干扰RNA抑制肝癌细胞株BEL7402 ICAM-1基因表达的研究

目的研究小干扰RNA（siRNA）抑制肝癌细胞系BEL7402基因ICAM-1表达的可行性。方法根据ICAM-1基因设计shRNA片断，构建pGenesil-1／ICAM-1表达载体，转染BEL7402细胞，实时定量PCR和免疫组织化学方法检测肝癌细胞中ICAM-1基因表达抑制情况。结果该实验已成功构建了针对ICAM-1基因的pGenesil-1／ICAM-1质粒。实时定量PCR检测结果显示，BEL7402细胞经特异性siRNA作用后ICAM-1基因的表达受抑制，其Ct值由28．8降低到21．6，免疫组织化学结果显示：RNA干扰（RNAi）能特异而有效地抑制BEL7402细胞中ICAM-1基因的表达。结论构建的pGenesil-1／ICAM-1重组质粒能有效抑制ICAM-1基因在肝癌细胞株BEL7402中的表达，为肿瘤的生物学治疗提供了新的方法和材料。

Inhibitory effect of siRNA on ICAM-1 expression in liver cancer BEL7402 cells

Objective] To illustrate the feasibility of gene silence of ICAM-1 gene triggered by small interfering RNA(siRNA)in liver cancer BEL7402 cells.Methods] Plasmids pGenesil-1/ICAM-1 carried shRNA for ICAM-1 gene were transfected to BEL7402 cells by using lipofectamine TM2000.The expression rate of ICAM-1 gene was measured by real-time relative quantitative PCR and immunohistochemistry.Results] The recombinant plasmid pGenesil-1/ICAM-1 was constructed successfully.The result real-time of PCR revealed that the threshold cycle value was reduced form 28.8 to 21.6 after BEL7402 cell were inhibited by siRNA,Immunohistochemistry showed that pGenesil-1/ICAM-1 vector mediated ICAM-1 siRNA could inhibit the expression of ICAM-1 in BEL7402 cells.Conclusion] siRNA targeting ICAM-1 mRNA can specially suppress the expression of targeting gene in BEL7402 cells,which provides a new method and material to the biological therapy of cancer