Proto-oncogene Wnt-1
cytoplasm
The effect on cell growth by Wnt1 RNAi in human neuroblastoma SH-SY5Y cell line.
<h4>Purpose</h4>We tested the hypothesis that Wnt signaling pathways are critical to neuroblastoma development. Our objective was to explore the novel role that Wnt/beta-catenin plays in human neuroblastoma cell line SH-SY5Y, including detection of expression of wnt1 and beta-catenin in SH-SY5Y, and the morphological and proliferative changes after Wnt1 RNAi in SH-SY5Y.<h4>Methods</h4>PCR, Western blot and immunofluorescence technology were used to detect the expression of Wnt1 in human neuroblastoma SH-SY5Y cell line. RNAi technology was used to knock down the expression of Wnt1in SH-SY5Y. SiRNA targeting Wnt1 was transfected into SH-SY5Y cells by Lipofectamine2000. The protein expression of Wnt1 and beta-catenin were detected by Western blotting 48 h after transfection. The quantity and the morphologic changes of the cells were recorded under light microscope. The growth curve of SH-SY5Y cells after RNAi transfection was drawn through MTT assay.<h4>Results</h4>Wnt1 was expressed in human neuroblastoma SH-SY5Y cells. The SH-SY5Y cell was successfully transfected with siRNA targeting Wnt1 mediated by Lipofectamine in vitro. The proteins expression of Wnt1 and beta-catenin decreased after transfection with siRNA; the numbers of the cells were decreased, accompanying abundant floating and dead cells under the light microscope. SH-SY5Y cells transfected with siRNA targeting Wnt1 showed less viability.<h4>Conclusion</h4>Wnt1 and beta-catenin expressed in SH-SY5Y cells. Knockdown of endogenous wnt1 expression could result in cell death and inhibit cell growth. From our study, we suggest that the activated embryonal development-related wnt1/beta-catenin pathway might take part in the oncogenesis and growth of neural crest-derived neuroblastoma.