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Effect of recombinant plasmid pEGFP-AFP-hTNF on liver cancer cells (HepG2 Cells) in vitro when delivered by PEG-PEI/Fe₃O₄ nanomagnetic fluid

, : Effect of recombinant plasmid pEGFP-AFP-hTNF on liver cancer cells (HepG2 Cells) in vitro when delivered by PEG-PEI/Fe₃O₄ nanomagnetic fluid. Journal of the Formosan Medical Association 110(5): 326-335

Gene delivery into liver cancer cells has been a problem. This study aimed to understand the effect of using PEGI/Fe₃O₄ nanomagnetic fluid as a gene vector for liver cancer gene therapy. An AFP enhancer aids in the expression of tumor-specific foreign genes in AFP-producing cancer cells like HepG2 cells, and was utilized in the delivery method in this study. We constructed recombinant plasmid PEGFP-AFP-hTNFα, which was transfected into AFP positive HepG2 cells and AFP negative Hela cells by PEG-PEI/Fe₃O₄ nanomagnetic fluid. Fluorescence microscopy was used to evaluate the transfection rate of the hTNFα gene in the HepG2 cells 12 hours after transfection. Reverse transcription polymerase chain reaction (RT-PCR) and western blot were used to detect expression of hTNFα gene in the HepG2 cells 48 hours after transfection. Methyl thiazolyl tetrazolium (MTT) assay was used to evaluate the inhibitory effect of hTNFα on the proliferation of HepG2 cells. Flow cytometry was used to analyze the apoptosis of HepG2 cells. Plasmid PEGFP-AFP-hTNFα delivered by PEG-PEI/Fe₃O₄ nanomagnetic fluid was successfully transfected into HepG2 cells and expressed in the HepG2 cells. The transfection efficacy of hTNFα gene delivered by PEG-PEI/Fe₃O₄ nanomagnetic fluid was superior to that of hTNFα gene delivered by lipofectamine in HepG2 cells. RT-PCR and western blot demonstrated that hTNFα gene was expressed in HepG2 cells that were transfected with complexes of nanomagnetic fluid/PEGFP-AFP-hTNFα. MTT and flow cytometry showed that the hTNFα gene markedly exerted a cell killing effect. PEG-PEI/Fe₃O₄ nanomagnetic fluid successfully transfected PEGFP-AFP-hTNFα into HepG2 cells and induced expression of hTNFα gene in the HepG2 cells, thus showing promise as a gene vector for liver cancer gene therapy. Furthermore, an AFP enhancer can specifically increase the expression of target genes in cells positive for AFP.


PMID: 21621154

DOI: 10.1016/S0929-6646(11)60049-1

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Yin, J.Q.; Wan, Y., 2002: RNA-mediated gene regulation system: now and the future (Review). The discovery of RNA-mediated gene regulation system has brought human beings another big surprise on the wisdom and power of nature. Nature can not only create genetic codes with 4 nucleotides and produce proteins with 21 amino acids, but also co...

Rummens, K.; Van Herck, E.; Van Bree, R., 2000: Dietary calcium and phosphate restriction in guinea-pigs during pregnancy: fetal mineralization induces maternal hypocalcaemia despite increased 1a,25-dihydroxycholecalciferol concentrations. The British Journal of Nutrition 84(4): 5-504

Prenner, K., 1969: Indication for amputation in patients with disorders of the peripheral arterial circulation. Munchener Medizinische Wochenschrift 111(42): 2150-2154

Esper, J.; Bosshard, A.; Schweingruber, F.H.; Winiger, M., 1995: Tree-rings from the upper timberline in the Karakorum as climatic indicators for the last 1000 years. Climatic sensitivity, mid-term fluctuations and tree reaction responsible for the growth effect of atmospheric CO2 enrichment were analysed using the width of growth rings of 69 juniper (Juniperus) trees in Karakorum, northern Pakistan. The skelet...

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Dietz, P.; Böhm, R.; Strauch, D., 1980: Experimental studies of the effectiveness and compatibility of formaldehyde gas as well as peracetic acid and hydrogen peroxide aerosols. Zentralblatt für Veterinarmedizin. Reihe B. Journal of Veterinary Medicine. Series B 27(4): 268-279

Nilsson, A., 1962: Estrogenic activity of some isoflavone derivatives. Five isoflavones, biochanin A, genistein formononetin, daidzeln and 5-7-dihydroxyisoflavone were synthesized. Their estrogenic activity was assayed by the mouse uterine weight method under different administration routes. Daidzein, in contrast to...

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Eckmann, L.; Resta Lenert, S.; Kagnoff, M.F.; Barrett, K.E., 1999: Butyrate-induced differentiation of intestinal epithelial cells is accompanied by decreased responsiveness to prostaglandin E2. Gastroenterology 116(4 PART 2): A876, April