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通过观察不同营养状况下NGF诱导PC12细胞发生周期阻滞过程中p53蛋白水平的变化,探讨p53在PC12细胞周期阻滞中可能的作用机制.用流式细胞术检测细胞周期;Westernblot检测p53和p21WAF1/CIP蛋白水平.结果显示1%FBS(FatalBovineSerum)和50μg/LNGF(NerveGrowthFactor)均可诱导PC12细胞发生细胞周期阻滞.在10%FBS+50μg/LNGF处理的细胞中,p53和p21WAF1/CIP1均增高,而使用MEK抑制剂U0126(10μmol/L)可以抑制这一增高.在1%FBS处理的细胞中,p53水平增高,p21WAF1/CIP1却未见明显增高;进而加入50μg/LNGF作用1h后,p53显著降低,6h后再次升高,并持续至24h.可见p53在50μg/LNGF和1%FBS诱导的细胞周期阻滞中均发挥作用,但作用机制可能不同.
To investigate the possible mechanism of p53 in the cell cycle arrest of PC12 cells by observing the changes of p53 protein during the process of cell cycle arrest induced by NGF under different nutrition conditions.Cell cycle was detected by flow cytometry and Western blotting was used to detect the expression of p53 and p21WAF1 / CIP protein.The results showed that 1% FBS (FatalBovineSerum) and 50μg / LNGF (Nerve Growth Factor) could induce cell cycle arrest in PC12 cells.P53 and p21WAF1 / CIP1 both in 10% FBS + 50μg / , While MEK inhibitor U0126 (10μmol / L) could inhibit this increase.When the cells were treated with 1% FBS, the level of p53 increased but the level of p21WAF1 / CIP1 was not increased obviously. p53 was significantly decreased and then increased again after 6h, and continued to 24h.We can see that p53 play a role in 50μg / LNGF and 1% FBS-induced cell cycle arrest, but the mechanism may be different.