短发夹RNA对肝癌细胞细胞间粘附分子-1基因表达的影响

目的利用小片段干扰RNA（siRNA）在肝癌细胞内诱导RNA干扰（RNAi），抑制细胞间粘附分子-1（ICAM-1）基因表达，在体外进行RNAi对肝癌治疗的实验研究。方法根据ICAM-1基因设计shRNA片断，构建pGenesil-1／ICAM-1表达载体，转入E．coli菌，并用Lipofectamine tm2000转染HepG2细胞，通过MTT方法检测细胞活性状态，应用免疫组织化学的方法检测肝癌细胞中ICAM-1基因蛋白质表达抑制情况。结果本实验已成功构建了针对ICAM-1基因的pGenesil-1／ICAM-1质粒，免疫组织化学结果显示：RNA干扰（RNAi）能特异而有效地抑制HepG2细胞中ICAM-1基因的表达。结论构建的pchlesd-1／ICAM-1重组质粒能有效抑制ICAM-1基因在肝癌细胞株HepG2中的表达和瘤细胞增殖。

Inhibition of ICAM-1 gene expression in liver cancer cells by shRNA

Objective To investigate the hepatocareinoma therapy by RNA interference,we used siRNA to inhibit ICAM-1 gene expression in vitro.Methods Plasmids pGenesil-1/ ICAM-1carried shRNA for ICAM-1 gene were transfected to HepG2 cells by using Lipofectamine TM2000.The mortality rate of HepG2 cells was determined by MTT staining,the protein level of ICAM-1 in the culture supernatant was measured with a enzyme-linked immunosorbent assay(ELISA).Results The recombinant plasmid pGenesil-1/ ICAM-1was constructed successfully.ELISA showed that pGenesil-1/ ICAM-1 vector mediated ICAM-1 siRNA could inhibit the expression of ICAM-1 in HepG2 cells.Conclusion siRNA targeting ICAM-1 mRNA can specially suppress the expression of targeting gene in HepG2 cells and cell growth.