ErbB3/Her3基因重组慢病毒的构建及应用*
Construction and application of recombinant ErbB3/Her3 lentiviral

目的:构建慢病毒Lenti-Her3,探索ErbB3/Her3在p44通路中的作用。方法:将ErbB3/Her3基因的两个片段Her3-1和Her3-2分段克隆至pCDH-CMV-MCS-EF1-Puro,构建慢病毒载体pCDH-Her3,然后将其包装成慢病毒Lenti-Her3; 用Lenti-Her3感染经p44-shRNA沉默的A549细胞,细胞计数法检测细胞的生长情况,Western blot法检测 P44、Her3蛋白的表达。结果:转染p44-shRNA的A549细胞P44蛋白不表达,Her3蛋白表达显著降低,细胞计数显著降低; Lenti-Her3感染后,p44沉默的A549细胞Her3蛋白表达显著增强,细胞计数有所升高(P＜0.05),但未达到正常水平。结论:成功构建了重组慢病毒Lenti-Her3; p44可部分通过Her3促进肺癌A549细胞的生长,可能成为肺癌药物研究的新靶点。
Aim: To construct lentiviral Lenti-Her3 and identify ErbB3/Her3 function in p44 pathway.Methods: Her3-1 and Her3-2 fragments of ErbB3/Her3 gene were cloned in turn into pCDH-CMV-MCS-EF1-Puro to generate pCDH-Her3,and then was packaged to obtain Lenti-Her3. Lenti-Her3 was infected in A549 cells which had been transfected with p44-shRNA, and then expressions of P44 and Her3 proteins were detected by Western blot, cell growth was detected through cell counting.Results: The A549 cells transfected with p44-shRNA had no expression of P44 protein, low expression of Her3 protein, and low cell number counting. The expression ofHer3 protein in the A549 cells transfected with p44-shRNA and infected with Lenti-Her3 was improved, and cell number counting was increased, but did not reach the normal level.Conclusion: The lentiviral Lenti-Her3 has been successfully constructed; p44 could promote lung cancer cell growth partially by Her3 pathway, which may be a potential therapeutic target for lung cancer