Abstract:

Abstract:Objective
To construct the hTERT promoter-driven TRAIL expression vector and explore its inhibitory effects on the proliferation of hepatoma SMMC7721 cells and HL7702 hepatocytes. Methods The recombinant plasmids pshuttle-TRAIL and pshuttle-hTERT-TRAIL were constructed with molecular biological methods. They were confirmed correctly by endonucleases digestion and PCR identification,and transfected into SMMC7721 and HL7702 cells through liposome,meanwhile,the pcDNA3.1+-GFP was transfected to detect the transfection efficacy by fluorescence microscopy. MTT was used to detect the effects of the recombinant plasmids on cell proliferation in transfected cells. Results The recombinant plasmid pshuttle-TRAIL and pshuttle-hTERT-TRAIL were constructed successfully. And the transfection efficacy was highest by fluorescence microscopy when the ratio of plasmid to liposome was 1∶3. As compared with control group, the inhibitory rates of SMMC7721 cells from 16 h after transfection in pshuttle-hTERT-TRAIL group was increased significantly (P<0.001);and as compared with pshuttle-TRAIL group,the inhibitory rates of SMMC7721 cells from 24 h after transfection in pshuttle-hTERT-TRAIL group was increased significantly (P<0.05). However,as compared with control group,the inhibiory rates of HL7702 cells in recombinant plasmids groups did not change significantly. Conclusion hTERT promoter-driven TRAIL could significantly inhibit the hepatoma cell proliferation,especially superior to that in pshuttle-TRAIL,however,no effects on hepatocyte proliferation.

Key words: TNF-related apoptosis inducing ligand;hTERT promoter;tumor targeting therapy;cell proliferation