Establishment and characterization of DNA pol β knockout human esophageal carcinoma cell line EC9706

To construct a DNA polymerase beta gene knockout model in human esophageal carcinoma cell EC9706 by homologous recombination for investigating its biological characterization and sensitivity upon damaging factors or chemotherapeutics. Methods: Based on the homologous recombination principle, the gene targeting vector was constructed to delete pol beta gene. The vector was introduced into esophageal carcinoma cellline EC9706 by electroporation. PCR, RT-PCR and Western blot were used to detect the expression of pol beta gene at DNA, mRNA and protein level in pol beta knockout EC9706 cell. Flow cytometry and MTT were used to detect cell cycle and cell growth velocity. The sensitivity of the gene targeting cell line upon oxydizing agent and chemotherapeutics were detected by trypan blue anti-dyeing method. Results: In the targeting cell line, the DNA, mRNA and protein expression of pol beta can not be detected and its biological characterization has marked disparation compared with the normal EC9706 cell. Conclusion: The pol beta gene knockout EC9706 cell line was constructed successfully. It may lay a foundation for the further study of pol beta gene. [Life Science Journal. 2010; 7(2): 13-18] (ISSN: 1097-8135).