Construction of TRAIL eukaryotic expression vector driven by the hTERT promoter and apoptotic effect on nasopharyngeal carcinoma cell line CNE-2 / 中国耳鼻咽喉头颈外科

ABSTRACT

OBJECTIVE To explore the effect of gene therapy on the nasopharyngeal carcinoma (NPC) cell line CNE-2 by using the combination of TRAIL gene with hTERT promoter. METHODS Total RNA was ex-tracted from PBL (peripheral blood lymphocytes)whose proliferation had been stimulated. TRAIL gene with interleukin 2 signal peptide gene was amplified by RT-PCR and cloned into the vector pGL3-181hTE down-stream to the RT promoter to form an eukaryotic vector. The vector was transfected into CNE-2 cells and HL-7702 cells through lipofection. Flow cytometry (FCM), agarose gel electrophoresis and cell morphology were used to examine the cell apoptosis. RESULTS In tranfected nasopharyngeal carcinoma cells CNE-2, FCM analysis showed that apoptotic peak appeared before G1 phase. A ladder-like pattern of DNA fragmentation appeared upon agarose gel electrophoresis. Many cells exhibited apoptotic changes such as cell shrinkage , nulear condensation , and nuclear fragmenta-tion under transmission electron microscope (TEM). CONCLUSION The recombinant eukaryotic ex-pression vector for TRAIL gene driven by hTERT pro-moter was successfully constructed and shown to induced apoptosis in CNE-2 cells. The results suggest that TRAIL may be a promising target for gene therapy of NPC.