The Influence Of Death Receptor6on The Human Neuroblastoma Cell Growth

ObjectiveThe purpose of this paper was to construct eukaryotic expressing vector ofpTK643-DR6, establish the human neuroblastoma cell line stably expressingDR6, and study the effect of DR6on cell proliferation.MethodsThe full-length DR6cDNA fragment was amplified by PCR from eukaryoticexpressing plasmid FG30-DR6and cloned into the lentiviral vector expressionplasmid pTK643(with green fluorescent protein (GFP) gene). After digestionand sequencing, the recombinant pTK643-DR6vector was infected intoSH-SY5Y cells by lentivirus. A stably-transfected SH-SY5Y cell line wasestablish. Expression of DR6was identified by Flow Cytometry and WesternBlot. The influence of DR6on SH-SY5Y cells was detected by Growth Curve.ResultsThe pTK643-DR6expression vector was successfully constructed. Astably-expressing DR6SH-SY5Y cell line was obtained via lentiviral infection,which was displayed by Flow Cytometry and Western Blot. From the result ofGrowth Curve, high level expression of DR6promote the death of SH-SY5Ycells, while low expression has little effect.ConclusionsDR6stably expressing SH-SY5Y cell line was successfully obtained,which may provide a cell model and experimental data for further investigationon DR6function and impaction of neuro-oncology. The high level expression ofDR6affect the proliferation of SH-SY5Y cells, and the influence of low level expression of DR6on SH-SY5Y cells is not obvious, of which the exactmechanisms still need further study.