%A CHEN Sheng, GU Shuo, XU Min, et al %T Mechanism of effects of rapamycin on growth inhibition and inducing apoptosis of human neuroblastoma cell line %0 Journal Article %D 2014 %J Journal of Shanghai Jiao Tong University (Medical Science) %R 10.3969/j.issn.1674-8115.2014.04.016 %P 481- %V 34 %N 4 %U {https://xuebao.shsmu.edu.cn/CN/abstract/article_10286.shtml} %8 2014-04-28 %X
Objective To investigate the effects of rapamycin on growth inhibition and inducing apoptosis of human neuroblastoma cell line SH-SY5Y and the target signaling pathway of phosphatidylinositol-3-kinase/protein kinase B/the mammalian target of rapamycin (PI3K/Akt/mTOR). Methods The rapamycin intervention group contained SH-SY5Y cells treated by the rapamycin of different concentrations (10, 15, 20 μmol/L). The negative control group contained SH-SY5Y cells by adding the same volumes of DMSO. The blank control group was also established. The CCK-8 assay, flow cytometry assay, and annexin Ⅴ-FITC/PI double staining were used to determine cell proliferation rate, cell cycle, and apoptosis rate of SH-SY5Y cells, respectively. Western blotting was performed to determine expressions and changes of phosphorylation level of cleaved caspase-3 and upstream and downstream molecules of signaling pathway of PI3K/Akt/mTOR, including PI3Kp85, Akt, mTOR, and 4E-BP1. Results Compared to the negative control group and blank control group, the cell growth rate of rapamycin intervention group decreased significantly (P<0.05 or P<0.01). Cell cycle analysis further showed that rapamycin could arrest the cell cycle at G0/G1 phase (P<0.05). Early and late apoptosis in SH-SY5Y cells treated by rapamycin were evident through annexin Ⅴ-FITC/PI staining assay (P<0.01). The results of Western blotting showed that the expression level of cleaved caspase-3 of rapamycin intervention group was higher than that of the negative control group and blank control group; the activities of upstream and downstream molecules of signaling pathway of PI3K/Akt/mTOR were inhibited; and the phosphorylation levels of PI3K, Akt, mTOR, and 4E-BP1 were significantly decreased. Conclusion The rapamycin can inhibit the cell growth and induce the apoptosis of human neuroblastoma cell line SH-SY5Y. The mechanism may be relevant to the suppression of signaling pathway of PI3K/Akt/mTOR.