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Research on p85a Gene in Migration of Human Neuroblastoma SY5Y Cell
Chen Linhua, Jing Xia, Wang Lihua, Wang Jiao*
School of Ophthalmology and Optometry, Eye Hospital, Wenzhou Medical University; State Key Laboratory of Ophthalmology, Optometry and Visual Science, Wenzhou 325027, China
Abstract: By interfering the expression of p85a gene， we investigated the function of p85a on migration， and the effection with mRNA and protein expression in human neuroblastoma SY5Y cell. First， the RNAi plasmid (si-p85a) and overexpression plasmid (OE-p85a) were constructed for p85a gene. Si-p85a and OE-p85a were transferred separately into cultured human neuroblastoma SY5Y cell. After 48 hours， Real-time PCR and Western blot analysis were carried out to detect the mRNA and protein expression level of p85a gene. The cell cycle was evaluated by flow cytometry. SY5Y cell migration was examined by wound-healing assay. The results of Real-time PCR showed that the p85a gene expressed differentially compared with the MOCK control. The expression level of si-p85a group was reduced by 58%. Conversely， the expression of OE-p85a group was significantly increased， which was 22.84 times that of the control group (P<0.05). Western blot indicated that p85a protein expression was decreased by transfection of si-p85a plasmid. On the other hand， transfection of OEp85a plasmid increased p85a protein expression. Wound-healing assay showed that cell migration was promoted in OE-p85a plasmid transfected SY5Y cell. Consistently， migration was retarded in si-p85a plasmid transfected SY5Y cell (P<0.05). During SY5Y cell growth process， p85a plays an important role in promoting the migration of SY5Y cells.
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