论文部分内容阅读
背景与目的研究KDR启动子转录调控双自杀基因系统(CDglyTK)对人大细胞肺癌细胞L9981的杀伤作用。方法应用分子生物学技术克隆KDR基因的启动子KDRp,构建KDR启动子调控的双自杀基因(CDglyTK)真核表达质粒pcDNA3-KDRp-CdglyTK。并将其导入L9981和人肝癌细胞HepG2中。给予不同的前药(GCV和/或5-FC)处理后,应用MTT法检测各组细胞的存活率。并应用流式细胞仪检测各组细胞的细胞周期和凋亡。结果成功的克隆KDR启动子和构建pcDNA3-KDRp-CdglyTK质粒。pcDNA3-KDRp-CdglyTK转染后,L9981细胞中均可检测到CD和TKmRNA,而HepG2细胞中则无。联合应用5-FC+GCV处理对转双自杀基因细胞(L9981)的杀伤作用显著高于单独应用5-FC或GCV(P<0.05),且二者显示了良好的药物协同作用。结论KDR基因启动子调控双自杀基因系统可以靶向性杀伤人肺癌细胞。
BACKGROUND & AIM: To study the killing effect of KDR promoter double transcription and suicide gene system (CDglyTK) on human large cell lung cancer cell line L9981. Methods The KDR gene promoter KDRp was cloned by molecular biology technique and the eukaryotic expression vector pcDNA3-KDRp-CdglyTK was constructed by the double suicide gene (CDglyTK) regulated by KDR promoter. And introduced into L9981 and human hepatoma HepG2 cells. After treatment with different prodrugs (GCV and / or 5-FC), the survival rate of each group of cells was measured by MTT method. Flow cytometry was used to detect cell cycle and apoptosis in each group. Results The KDR promoter was successfully cloned and the pcDNA3-KDRp-CdglyTK plasmid was constructed. After transfection with pcDNA3-KDRp-CdglyTK, both CD and TK mRNA were detected in L9981 cells, but not in HepG2 cells. The combination of 5-FC + GCV treatment was significantly more potent than 5-FC or GCV (P <0.05) in killing dual-suicide gene cells (L9981), and both showed good synergistic effects. Conclusion The double suicide gene system regulated by KDR gene promoter can specifically kill human lung cancer cells.