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Effect of N-terminal Fragment Fusion Protein of Polycystin-1 on Cell Cycle and Its Regulatory Gene in Rat Mesangial Cell
Tian-Jun Guan, Chang-Lin Mei*, Tian-Mei Sun, Li-Li Fu, Hai-Dan Zhao1, Hou-An Cai
Department of Nephrology, Changzheng Hospital, Second Military Medical University, Shanghai 200003, China; 1Division of Nephrology, Beijing Sea Army Hospital, Beijing 100037, China
Abstract: To investigate the effect of N-terminal fragment (LRR-WSC) fusion protein of polycystin-1 (PC-1NF) on cell cycle and its regulatory gene in rat mesangial cell and explore its mechanism. Rat mesangial cells were treated with serial concentration of PC-1NF in vitro. The proliferation of rat mesangial cells was measured by Brdu-ELISA. Cell cycle was analyzed by flow cytometry. The mRNA expression of cell cycle regulatory gene cyclin D1 and p21WAF1 was detected by real-time fluorescence quantitative PCR. Micro-gram per milliliter of PC-1NF could effectively inhibit the proliferation of mesangial cells with dose- and time-dependent correlation to some extent， and cause G0/G1 phase arrest. After treatment with 4 µg/ml PC-1NF， the mRNA level of cyclin D1 was significantly decreased [(2.44±0.27)×104 copies per million GAPDH] compared with that in the control [(4.10±0.32)×104 copies per million GAPDH]. However， the mRNA level of p21WAF1 was significantly increased [(3.73±0.46)×102 copies per million GAPDH] compared with that in the control [(0.85±0.18)×102 copies per million GAPDH]. N-terminal fragment fusion protein of polycystin-1 could effectively inhibit mesangial cell proliferation. The effect could be mediated by down-regulated cyclin D1 and up-regulated p21WAF1 mRNA expression， subsequently blocking cells passing through G1-S checkpoint.
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