论文部分内容阅读
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pGenesil-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。
OBJECTIVE: To study the effects of RNAi on the treatment of hepatocellular carcinoma (HCC) in vitro using small interfering RNA (siRNA) to induce RNAi and ICAM-1 gene expression in hepatoma cells. Methods According to the designed shRNA fragment of ICAM-1 gene, pGenesil-1 / ICAM-1 expression vector was constructed and transfected into E.coli. HepG2 cells were transfected with Lipofectamine tm2000. The cell viability was detected by MTT assay. Immunohistochemistry Inhibition of ICAM-1 gene protein expression in hepatocellular carcinoma cells. Results The pGenesil-1 / ICAM-1 plasmid targeting ICAM-1 gene was successfully constructed in this study. The results of immunohistochemistry showed that RNA interference (RNAi) could specifically and effectively inhibit the expression of ICAM-1 gene in HepG2 cells. Conclusion The recombinant plasmid pGenesil-1 / ICAM-1 can effectively inhibit the expression of ICAM-1 gene in HepG2 cells and proliferation of tumor cells.