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- Title
The effect on cell growth by Wnt1 RNAi in human neuroblastoma SH-SY5Y cell line.
- Authors
Lihong Zhang; Kai Li; Zhibao Lv; Xianmin Xiao; Jicui Zheng; Zhang, Lihong; Li, Kai; Lv, Zhibao; Xiao, Xianmin; Zheng, Jicui
- Abstract
<bold>Purpose: </bold>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.<bold>Methods: </bold>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.<bold>Results: </bold>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.<bold>Conclusion: </bold>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.
- Subjects
NEUROBLASTOMA; NERVOUS system tumors; GROWTH factors; CELL lines; IMMUNOFLUORESCENCE; CELL physiology; COMPARATIVE studies; CYTOSKELETAL proteins; GENES; RESEARCH methodology; MEDICAL cooperation; PROTEINS; RESEARCH; RNA; WESTERN immunoblotting; FLUORESCENCE in situ hybridization; EVALUATION research; FLUOROIMMUNOASSAY
- Publication
Pediatric Surgery International, 2009, Vol 25, Issue 12, p1065
- ISSN
0179-0358
- Publication type
journal article
- DOI
10.1007/s00383-009-2481-0