Endostatin gene therapy for liver cancer by a recombinant adenovirus delivery

摘要： <正> AIM:To investigate the expression of adenovirus-mediatedhuman endostatin(Ad/hEndo)gene transfer and its effecton the growth of hepatocellular carcinoma(HCC)BEL-7402xenografted tumors.METHODS:Immunohistochemistry analysis with an anti-endostatin antibody was preformed to detect endostatinprotein expression in HCC BEL-7402 cells infected withAd/hEndo.MTT assay was used to investigate the effects ofAd/hEndo on proliferation of human umbilical vein endothelialcells(HUVEC).Intra-tumoral injections of 1×10~9 pfu Ad/hEndowas given to treat BEL-7402 xenografted tumors in nudemice once weekly for 6 wk.Mice received injections ofAd/LacZ and DMEM were regarded as control groups.Afterintra-turmoral administration with Ad/hEndo,the endostatinmRNA expression in tumor tissue was analyzed by Northernblotting,and plasma endostatin levels were determined usingenzyme-linked immunosorbent assay(ELISA).RESULTS:High level expression of endostatin gene wasdetected in the infected HCC BEL-7402 cells.Ad/hEndosignificantly inhibited HUVEC cell proliferation by 57.2% ata multiplicity of infection(MOI)of 20.After 6-weektreatment with Ad/hEndo,the growth of treated tumorswas inhibited by 46.50% compared to the Ad/LacZcontrolgroup(t=2.729,P<0.05)and by 48.56% compared to theDMEM control group(t=2.485,P<0.05).The ratio of meantumor volume in treated animals to mean tumor volume inthe control animals(T:C ratio)was less than 50% after 24 dof treatment.Endostatin mRNA in tumor tissue was clearlydemonstrated as a band of approximately 1.2 kb,whichwas the expected size of intact and functional endostatin.Plasma endostatin levels peaked at 87.52±8.34 ng/mLat d 3 after Ad/hEndo injection,which was significantlyhigher than the basal level(12.23±2.54 ng/mL).By d 7,plasma levels dropped to nearly half the peak level(40.34±4.80 ng/mL).CONCLUSION:Adenovirus-mediated human endostatin gene can successfully express endogenous endostatin invitro and in vivo,and significantly inhibit the growth ofBEL-7402 xenograffed liver tumors in nude mice. ... （共5頁）