Cloning of secreted hTRAIL tumor-specific vector and expression in laryngeal carcinoma cell line Hepa2

    CHENG Shiª²Yin1 , ZHANG Huiª²Zhong2,CHEN Jianª²Qiu1, ZHANG Li1,HAN Mingª²Kun1

    1Department of Otorhinolaryngology, 2Center Laboratory ,Tangdu Hospital, Fourth Military Medical University, Xi¬ðan 710038, China

    ¡¾Abstract¡¿ AIM£º To clone human soluble TRAIL cDNA and construct its eukaryotic expression vector to prepare a condition for targeted gene therapy. METHODS£º The total RNA was extracted from human peripheral blood lymphocytes. TRAIL gene with interleukin 2 signal peptide was amplified by RTª²PCR and cloned into the vector pGL3ª²181hTERT promoter downstream to form an eukaryotic vector. Expressing protein was identified by Western Blotting on laryngeal carcinoma cell line Hepa2. RESULTS£º Sequence of 613 bp cDNA was obtained. Identified by enzyme digestion and sequence analysis, the recombinant eukaryotic expression vector for TRAIL gene had been successfully constructed. The results of the Western Blotting showed the transfected Hepa2 cell could express TRAIL protein. CONCLUSION£º Successful construction of eukaryotic expression vector pGL3ª²181 hTERT/TRAIL provides the possibility for gene therapy of tumor. ......