The effect on cell growth by Wnt1 RNAi in human neuroblastoma SH-SY5Y cell line.
OBJECTIVE
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.


METHODS
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.


RESULTS
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.


CONCLUSIONS
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.
Zhang
Lihong
L
Department of Pediatric Surgery, Children's Hospital of Fudan University, 399 Wang Yuan Road, 201102 Shanghai, People's Republic of China. likai2727@163.com
Li
Kai
K
Lv
Zhibao
Z
Xiao
Xianmin
X
Zheng
Jicui
J
D015153:Blotting, Western; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D015200:Fluoroimmunoassay; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D009447:Neuroblastoma / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D012329:RNA, Bacterial / Q000235:genetics*; D012334:RNA, Neoplasm / Q000096:biosynthesis / Q000235:genetics*; D051155:Wnt1 Protein / Q000096:biosynthesis / Q000235:genetics*; D051176:beta Catenin / Q000096:biosynthesis / Q000235:genetics
C042027:RNA I; D012329:RNA, Bacterial; D012334:RNA, Neoplasm; C495247:WNT1 protein, human; D051155:Wnt1 Protein; D051176:beta Catenin