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目的构建CD147慢病毒干涉载体,建立稳定下调CD147表达的人肺腺癌A549细胞系,观察下调CD147表达后人肺腺癌细胞增殖能力的变化。方法将携带不同特异性干涉序列的3个DNA片段分别插入干涉质粒pSicoR中,构建pSicoR-siCD147慢病毒干涉载体(pSicoR-siCD147-1,pSicoR-siCD147-2,pSicoR-siCD147-3)。分别将构建好的载体转染入293FT细胞中,进行慢病毒包装。建立稳定下调CD147表达的细胞系。利用CCK-8法检测细胞增殖能力的变化。结果与结论构建了pSicoR-siCD147慢病毒干涉载体,RT-PCR和实时PCR结果显示pSicoR-siCD147-2的干涉效率最高。建立了稳定下调CD147表达的人肺腺癌A549细胞系(A549-siCD147)。CCK-8法检测发现A549-siCD147细胞的增殖能力显著性下降(P<0.05),下调CD147的表达可以使人肺腺癌细胞的增殖能力受到明显抑制。
Objective To construct a CD147 lentiviral vector and establish a human lung adenocarcinoma A549 cell line stably downregulating CD147 expression and observe the changes of the proliferation of human lung adenocarcinoma cells after down-regulating the expression of CD147. Methods Three DNA fragments carrying different specific interference sequences were inserted into the interference plasmid pSicoR respectively to construct the pSicoR-siCD147 lentiviral vector (pSicoR-siCD147-1, pSicoR-siCD147-2, pSicoR-siCD147-3). The constructed vector was transfected into 293FT cells for lentivirus packaging. To establish a cell line that stably down-regulates CD147 expression. The changes of cell proliferation were detected by CCK-8 assay. Results and Conclusion The pSicoR-siCD147 lentiviral vector was constructed. The results of RT-PCR and real-time PCR showed that the interference efficiency of pSicoR-siCD147-2 was the highest. A human lung adenocarcinoma A549 cell line (A549-siCD147) stably down-regulated CD147 expression was established. The proliferation of A549-siCD147 cells was significantly decreased by CCK-8 assay (P <0.05). The down-regulation of CD147 expression could significantly inhibit the proliferation of human lung adenocarcinoma cells.