国产雷帕霉素对人淋巴瘤细胞Raji增殖的影响
Effect of Rapamycin in Proliferation of Human Burkitt Lymphoma Cells
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摘要: 目的 探讨国产雷帕霉素（宜欣可）对人淋巴瘤细胞株Raji细胞体外生长及对mTOR/p 70S6K信号通路的影响。方法MTT法检测不同浓度（0、1、5、10、20、40、50、100 nmol/L）国产雷帕霉素作用不同时间（24、48、72 h）对Raji 细胞增殖的影响。光学显微镜观察Raji细胞形态学变化。流式细胞仪测定国产雷帕霉素对Raji细胞周期分布和凋亡的影响。Western blot 方法检测国产雷帕霉素处理前后对Raji 细胞mTOR、p70S6K、p-p70S6K蛋白的影响。结果国产雷帕霉素对Raji细胞增殖有明显的抑制作用（不同浓度P<0.01或P<0.05），呈现明显的剂量-效应和时间-效应依赖关系。国产雷帕霉素明显抑制Raji细胞周期发展（P<0.05），但没有发生明显的凋亡（P>0.05）。0、10、50、100 nmol/L国产雷帕霉作用于Raji细胞的mTOR、p-p70S6K，其蛋白量随药物浓度增大而降低（P<0.05），p70S6K随药物浓度增大而升高（P<0.05）。结论人淋巴瘤细胞株Raji中存在mTOR/p70S6K信号通路激活状态，宜欣可可抑制该通路激活并通过阻滞细胞周期发展抑制Raji细胞增殖。Abstract: Objective To investigate rapamycin effects on growth inhibition and mTOR/p70S6K signaling pathway of human Burkitt lymphoma cell line Raji cells. Methods Proliferations of Raji cells under different of concentrations (0，1，5，10，20，40，50 and 100nmol/L) and different times (24，48 and 72 h) were investigated by MTT assay.Apoptosis and cell cycle were analyzed via flowcytometry.The morphological alterations were confirmed by the optical microscope.The expressions of mTOR，p70S6K，p-p70S6K proteins were examined by Western blot technique in the rapamycin-treated and untreated Raji cells. Results Rapamycin inhibited the proliferation of Raji cells at concentrations more than 5nmol/L(P＜0.05)，in a dose and time dependent manner.After treatment with rapamycin，the number of cells at S phase and G2/M phase was decreased gradually (P＜0.05)，but significantly increased at G0/G1 phase in dose and time dependent manners (P＜0.05).However，evident apoptosis did not observed in Raji cells.The expression of mTOR、p-p70S6K proteins was decreased gradually while the expression of p70S6K protein was increased in a concentration-dependent manner (P＜0.05). Conclusion mTOR/p70S6K signaling pathway was constitutively activated in Raji cells and Rapamycin inhibited Raji cell proliferation by arrest at G0/G1 phase and inhibition of mTOR/p70S6K signaling pathway.
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