Target delivery of lactose poly-L-lysine combined HSV-TK to human liver cancer cells / 中国医师杂志

ABSTRACT

Objective To explore a new molecular target for HSV-tk/GCV system in human liver cancer therapy.Methods The lactose and poly-L-lysine covalently linked compound(Lac-PLL) were prepared by using reductive amination methods and purified by using Sephadex G10 gel filtration.The value of n was determined by methods of phenol-vitriol colorimetry.The plasmid r-pAs16Dr was mixed with the conjugate to form a gene delivery complex named GlanPLL-r-pAs16Dr.The GlanPLL-r-pAs16Dr was transformed to different cell lines such as HepG2 and A549 to confirm the expression of RFP.The expression of HSV-tk was confirmed by RT-PCR.Cells with various concentrations of GCV were observed at different time points using MTT.Results The PLL modified by 34 Lac was obtained by using chemical synthesis.The RFP was expressed in HepG2 by 48h after transfection,and was not expressed in A549.The expression of HSV-tk was only detected in HepG2 using RT-PCR.The HepG2 transformed with GlanPLL-r-pAs16Dr was sensitive to GCV and the growth inhibiting rate was 70.5% with the treatment of low concentration of GCV(1mg/L) for 3 days.The A549 was not sensitive to GCV.Conclusion Lac-PLL,which is easy to prepare,is an efficient carrier for HSV-tk to be delivered to hepatoma cell lines by binding to ASGPR.