稳定低表达CD147的人肺腺癌细胞系的建立及对细胞增殖的影响  

Establishment of human lung adenocarcinoma cell line with stable low expression of CD147 and its effect on proliferation

作　　者：杨绍兴[1] 汤传昊[1] 王思涵[1] 宋三泰[1] 刘晓晴[1] 

机构地区：[1]军事医学科学院附属医院肺部肿瘤科,北京10007

出　　处：《军事医学》2012年第12期912-915,928,共5页Military Medical Sciences

基　　金：首都医学发展基金资助项目(2007-3042)

摘　　要：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 CD147 lentiviral RNA interference（RNAi） vector and human lung adenocarcinoma cell line A549 that is consistently down-regulated by CD147, and to observe its effect on proliferation of human lung adeno- carcinoma cells. Methods Three cloning DNA segments carrying different specific interfering sequences were inserted into lentivirus particles pSicoR respectively. CD147 lentiviral RNAi vectors （pSicoR-siCD147-1, pSicoR-siCD147-2 and pSi- coR-siCD147-3 ）were constructed. The pSicoR-siCD147 vector and pEGFP vector were transferred into 293FT cells respec- tively to produce a lentivirus equipment package. A stable cell line named A549-CD147 that could express CD147 lowly was established. The changes in cell proliferation were observed using CCK-8. Results and Conclusion The lenticiral RNAi vectors of CD147 （ pSicoR-siCD147 ） were successfully constructed. The expression of CD147 down-regulated by pSiocR- siCD147-2 lentiviral RNAi vector showed a higher interference efficiency than other groups detected with ：RT-PCR and real- time PCR. The human lung adenocarcinoma cell line with stable low expression of CD147 gene named A549-siCD147 was successfully established. The result of CCK-8 detection showed that the low expression of CD147 in A549 cells could significantly decrease cell proliferation （ P 〈 0.05 ）. Down-regulated expression of CD147 could obviously inhibit the proliferation of human lung adenocarcinoma cells.

关 键 词：CD147 慢病毒属 RNA干涉 肺肿瘤 细胞增殖 

分 类 号：R734.2[医药卫生—肿瘤]