雷帕霉素对骨肉瘤细胞MG63核干细胞因子表达的调节
Effect of Rapamycin on Nucleostemin Expression in Osteosarcoma MG63 Cells
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摘要: 目的 探讨雷帕霉素（rapamycin, RAPA）对骨肉瘤细胞MG63核干细胞因子（nucleostemin,NS）的调节及可能机制。 方法 反转录聚合酶链式反应（reverse transcription polymerase chine reaction,RT-PCR)和Western blot检测核干细胞因子（nucleostemin，NS）转录和蛋白表达，通过激酶抑制剂预处理找出可能参与RAPA调节NS的激酶；Western blot检测RAPA对此激酶活性的影响。 结果 MG63细胞中RAPA 在转录和蛋白表达水平均上调了NS表达。蛋白激酶B(Akt)抑制剂wortmannin预处理细胞逆转了RAPA对NS的上调作用。RAPA对Akt的总体活性无明显影响，但上调了第473位丝氨酸磷酸化Akt(phosphorylated Akt at Ser473, pAkt)的核转位。 结论 RAPA可能通过增强pAkt的核转位上调了MG63细胞中NS的表达。Abstract: Objective To investigate the regulation and possible mechanisms of rapamycin (RAPA) on nucleostemin (NS) expression in osteosarcoma MG63 cells. Methods After treatment with RAPA or in combination with various kinase inhibitors pre-treatment, the mRNA and protein expression of NS were determined by reverse transcription polymerase chine reaction (RT-PCR) and Western blot respectively. Effect of RAPA on the selected kinase was analysed by Western blot. Results RAPA up-regulated NS mRNA and protein expression and the protein kinase B (Akt) inhibitor prevented the effects of RAPA. RAPA did not affect the total Akt activity. However, the nuclear translocation of phosphorylated Akt at Ser473 (pAkt) was enhanced by RAPA. Conclusion RAPA possibly increases the expression of NS in MG63 cells through enhancing the nuclear translocation of pAkt.
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