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目的 :应用慢病毒介导的RNA干扰技术沉默胆管癌RBE细胞中人髓细胞白血病因子1相互作用蛋白(MLF1IP)基因的表达,探讨其对胆管癌细胞体外增殖的影响。方法 :设计合成针对MLF1IP靶基因序列的siRNA序列,进行重组慢病毒包装。与不含有MLF1IP基因si RNA序列的慢病毒分别转染胆管癌RBE细胞株。Real-time PCR检测MLF1IP基因的mRNA表达,MTT法检测细胞的增殖能力。结果 :成功构建了MLF1IP-sh RNA慢病毒质粒并进行慢病毒包装,Real-time PCR显示实验组MLF1IP基因的mRNA相对表达量为0.15±0.02明显高于对照组为1.01±0.12。MTT法检测实验组细胞的OD值对比对照组显著降低。结论 :慢病毒介导的RNA干扰技术能有效沉默RBE细胞中MLF1IP基因的表达,并能明显抑制胆管癌细胞RBE的体外增殖能力。
Objective: To silence the expression of human myeloid leukemia factor-1 interacting protein (MLF1IP) gene in RBE cells by lentivirus-mediated RNAi and investigate its effect on the proliferation of cholangiocarcinoma cells in vitro. Methods: The siRNA sequence targeting MLF1IP target gene was designed and synthesized and packaged by recombinant lentivirus. RBE cells were transfected with lentivirus without MLF1IP gene si RNA sequence respectively. The mRNA expression of MLF1IP gene was detected by Real-time PCR and the proliferation of cells was detected by MTT assay. Results: The lentiviral plasmid MLF1IP-sh RNA was successfully constructed and packaged with lentivirus. Real-time PCR showed that the relative mRNA level of MLF1IP gene in experimental group was 0.15 ± 0.02, which was significantly higher than that in control group (1.01 ± 0.12). The OD value of cells in experimental group detected by MTT method was significantly lower than that in control group. CONCLUSION: Lentivirus-mediated RNA interference can effectively silence the expression of MLF1IP gene in RBE cells and significantly inhibit the proliferation of RBE cells in vitro.