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Effects of Protein Phosphatase Inhibitor Okadaic Acid on Phosphorylation of Tau Protein in Human Neuroblastoma SK-N-SH Cells
Yan-Qi Chu, Wei Li1, Lan Zhang, Hou-Xi Ai, Yα-Li Li, Lin Li*
Department of Pharmacology, Xuanwu Hospital of Capital University of Medical Sciences, Beijing 100053, China;1Department of Neurology, Renmin Hospital of Henan Province, Zhengzhou 450000, China
Abstract: To investigate the effects of protein phosphatase (PP-1 and PP-2A) inhibitor okadaic acid (OA) on human neuroblastom SK-N-SH cells， and to determine the best dosage and duration of OA treatment in this cell model. The diverse doses of OA were incubated with SK-N-SH cells for different periods of time. The changes in cell morphology were observed by microscope. The phosphorylation and non-phosphorylation level of tau protein at Ser202 site and Ser404 site were detected with Western blotting method. Incubation of 10_160 nmol/L OA with SK-N-SH cells for 3_24 h induced dose- and time-dependent changes in cell morphology， in which the starting dose and time of OA effect were 10 nmol/L and 3 h respectively. When 10 nmol/L OA was incubated with SK-N-SH cells for 6_24 h， the expression of phosphorylated tau protein at Ser199/Ser202 site and Ser404 site was increased， the expression of non-phosphorylated tau at Ser202 site and Ser404 site was decreased， and the total tau protein did not show obvious changes. OA is a good tool drug for studying hyperphosphorylation of tau protein， and 10 nmol/L OA incubation with human neuroblastoma SK-N-SH cells for 6 h can be taken as the proper conditions for developing the cell model of tau hyperphosphorylation.
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