In vitro study on reversing the drug resistance of choriocarcinoma cell line JEG-3/VP16 by the expression of adenovirus-mediated hTNFα gene in human umbilical cord blood-derived mesenchymal stem cells
MA Yan
XIANG Yang
CHEN Yan
FENG Feng-zhi
WAN Xi-run
YANG Xiu-yu
Abstract：Objective To construct the replication-deficient recombinant adenoviruses vectors contained human TNFα gene and to investigate TNFα expression in human umbilical cord blood-derived mesenchymal stem cells(hUCB-MSCs).And to observe the drug-resistance changes of JEG-3/VP1 6 after co-culture with hUCB-MSCs transfected with adenovirusmediated hTNFα gene.Method Human TNF-α gene was amplified by PCR method from plasmid pCMV-SPORT6-TNF and cloned into shuttle vector pDC316-IRES-EGFP which was carried into 293 cells together with backbone plasmid pBHGlox_E1,3Cre by lipofectamine 2000 to obtain packed recombinant adenovirus Ad-TNFα.hUCB-MSCs were isolated,cultivated and identified in vitro.The resistance index of JEG-3/VP16 was determined by MTT test after co-culture with hUCB-MSCs transfected with hTNFα gene.Result (1)The packed recombinant adenovirus Ad-TNFα was identified using PCR and enzyme digestion.(2)The adenovirus vector was constructed successfully and high efficient expression of TNFα in hUCB-MSCs was detected.(3)MTT test showed that the resistance index of JEG-3/VP16 decreased after coculture with hUCB-MSCs transfected huTNF-α gene.Conclusion The drug resistance of JEG-3/VP16 can be reversed after co cultured with hUCB-MSCs transfected with hTNFα gene.
Keywords：mesenchymal stem cellstumor necrosis factorrecombinant adenovirus vectordrug-resistance
Publication Date：2010-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：3( 193-195 )