Apoptosis of pancreatic carcinoma mediated by a retroviral vector containing antisense human telomerase gene / 基础医学与临床

ABSTRACT

Objective To study the apoptosis of pancreatic cancer cells induced by a retroviral vector containing antisense human telomerase RNA gene. Methods Sense and antisense telomerase RNA genes were transfected into Can-pan-2 cell line by electroporation,and selected with G418 to get stable transfection and retrovirus yielding cell lines. We transfected Can-pan-2 cell with concentrated retroviral supernatant,and selected with G418 to get stable transfection cell lines,The expression of the aim gene in Can-pan-2 cell was confirmed through PCR,while TRAP-PCR-ELISA was used to determine the telomerase activity,and immunofluorescence was adopted to determine reproductive activity of the infected cells,flow cytometry(FCM)was used to detect cell apoptosis.ResultsThe titer of sense and antisense recombinant retrovirus reached 0.95?106 CFU/mL and 1.1?106 CFU/mL respectively,PCR demonstrated that the telomerase RNA genes were successfully intergrated into the target cells’ genome,The cell growth and telomerase activity of Can-pan-2 were significantly inhibited and apoptosis occurred after antisense virus infection. Conclusion Antisense human telomerase RNA may inhibite telomerase activity of pancreatic cancer cells and promote apoptosis of the cells.