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目的研究慢病毒介导的髓细胞白血病基因(Mcl-1)短发夹状RNA(shRNA)干扰对淋巴瘤细胞系SNK-6增殖和凋亡的影响。方法设计以Mcl-1为靶点的shRNA并构建携带此shRNA的慢病毒载体,转染淋巴瘤细胞系SNK-6,荧光定量PCR(qPCR)及Western blot方法检测病毒感染前后Mcl-1 mRNA及蛋白表达变化,四甲基偶氮唑盐法(MTT法)和流式细胞仪分析细胞增殖和凋亡变化的情况。结果成功构建Mcl-1-shRNA慢病毒表达载体,并可有效感染淋巴瘤细胞株SNK-6,感染后SNK-6细胞Mcl-1基因的mRNA水平及蛋白水平表达明显下调,MTT法检测显示慢病毒介导的Mcl-1基因shRNA干扰与对照病毒组相比可明显抑制SNK-6细胞增殖[(31.6±3.3)%vs(5.8±2.7)%,P<0.01],流式细胞仪测定感染后细胞凋亡明显高于对照病毒组[(28.9±2.1)%vs(5.3±1.5)%,P<0.01]。结论慢病毒介导的Mcl-1基因shRNA干扰技术可特异性阻断SNK-6细胞Mcl-1基因的表达,抑制细胞的增殖并促进细胞的凋亡。
Objective To investigate the effects of lentivirus mediated short hairpin RNA (Mcl-1) shRNA on the proliferation and apoptosis of lymphoma cell line SNK-6. Methods shRNA targeting Mcl-1 and lentiviral vector carrying this shRNA were designed and transfected into lymphoma cell line SNK-6. Quantitative real-time quantitative PCR (qPCR) and Western blot were used to detect the expression of Mcl-1 mRNA and Protein expression changes, MTT assay and flow cytometry analysis of cell proliferation and apoptosis changes. Results The Mcl-1-shRNA lentiviral vector was successfully constructed and effectively infected lymphoma cell line SNK-6. The mRNA and protein levels of Mcl-1 gene in SNK-6 cells were significantly down-regulated Virus-mediated Mcl-1 gene shRNA interference significantly inhibited the proliferation of SNK-6 cells compared with the control group [(31.6 ± 3.3)% vs (5.8 ± 2.7)%, P <0.01] Cell apoptosis was significantly higher than that of the control group [(28.9 ± 2.1)% vs (5.3 ± 1.5)%, P <0.01]. Conclusion Lentivirus-mediated Mcl-1 gene shRNA interference can specifically block the expression of Mcl-1 gene in SNK-6 cells, inhibit cell proliferation and promote apoptosis.