The mechanism of SW480 cell line death caused by transfection of G1CEACDNa gene / 中国普通外科杂志

ABSTRACT

Objective To study the mechanism of SW480 cell line death caused by transfection of retroviral vector-G1CEACDNa.Methods Plasmid G1CEACDNa was transferred into the SW480 cell line using liposomes method. RT-PCR was performed to examin the expression of CD gene indrectly.The cell growth curve was measured by means of cell counting. When the CD+SW480 cells were exposed to 5-FC (1mmol/L), the growth inhibition rate and the "bystander effect" were detected by MTT method.The ultrastructure was observed by electron microscope.Apoptosis was verified by flow cytometer .Results The product of RT-PCR showed a band at 1.5kb on the photo of electrophoresis. The growth of CD+ SW480 cells was inhibited 24h after administrating 5-FC,and the inhibition rates at 72h,96h,120h were 30.0%,50.0% and 80.0%,respectively.Apoptosis cells in different phases and apoptotic bodies in the field of electron microscope were observed. Meanwhile ,a few cells showed necrosis.Flow cytometer verified that a few cells appearred apoptosis 48h after exposed to 5-FC (1mmol/L), the apoptosis rate and the necrosis rate at 72h,96h were 20.2%,30.7% and 19.6%,21.1% respectively.Conclusions The death mechanism of SW480 cells transfected with G1CEACDNa followed by 5-FC treatment includes both necrosis and apoptosis.Apoptosis is possibly the bystander effect.