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目的:观察对放射敏感的早期生长反应基因-1(early growth response-1,Egr-l)启动子调控单纯疱疹病毒胸苷激酶(herpes simplex virus thymidine kinase,TK)基因是否对胰腺癌细胞的杀伤作用有增敏效应。方法:以细菌内同源重组法构建含TK基因的腺病毒载体pAdEgr-1-TK,转染人胰腺癌细胞株PC-3,60Co-γ射线照射后,逆转录-聚合酶链反应(RT-PCR)半定量分析各不同照射剂量组TK基因的mRNA的表达。加入前药丙氧鸟苷(ganciclovir,GCV),MTT法检测其对胰腺癌细胞的杀伤作用。结果:经60Co-γ射线照射后,与对照组胰腺癌细胞(0.86±0.11)相比,前药GCV可明显提高对转染pAdEgr-1-TK胰腺癌细胞的杀伤效率(0.08±0.03)(P<0.001)。结论:由Egr-l启动子调控的TK自杀基因在γ射线作用下可以显著提高杀伤胰腺癌细胞的能力。
AIM: To investigate whether the radiation-sensitive early growth response-1 (Egr-1) promoter regulates the expression of herpes simplex virus thymidine kinase (TK) on pancreatic cancer cells Effect sensitization effect. Methods: The adenovirus vector pAdEgr-1-TK containing the TK gene was constructed by homologous recombination in bacteria and transfected into human pancreatic cancer cell line PC-3 and 60Co-γ-ray. Reverse transcriptase-polymerase chain reaction -PCR) semi-quantitative analysis of different irradiation dose TK gene mRNA expression. The prodrug ganciclovir (GCV) was added and its killing effect on pancreatic cancer cells was detected by MTT assay. Results: Compared with the control group, the GCV significantly increased the killing efficiency of pAdEgr-1-TK pancreatic cancer cells (0.08 ± 0.03) compared with the control group (0.86 ± 0.11) after 60Co-γ irradiation P <0.001). Conclusion: The TK suicide gene regulated by Egr-1 promoter can significantly enhance the ability of killing pancreatic cancer cells under the action of γ-rays.