摘要
目的探讨PI3K/Akt/mTOR信号通路在表阿霉素抑制人T细胞淋巴瘤细胞株Jurkat细胞增殖和诱导凋亡中的作用。方法用0、1.25、2.5、5、10μmol/L表阿霉素和0、0.25、0.5、1、2μmol/L PI3K/mTOR双重抑制剂(NVP-BEZ235)对Jurkat细胞作用48h后,CCK-8试剂盒检测Jurkat细胞株增殖抑制情况;采用AnnexinⅤ/PE双染法流式细胞术检测上述药物作用Jurkat细胞48h的凋亡率以及5μmol/L表阿霉素和2μmol/L NVP-BEZ235单独及联合作用Jurkat细胞0、12、24、36、48h的凋亡率;Western blotting法检测5、10μmol/L的表阿霉素作用Jurkat细胞0、6、12、24、48h,以及5μmol/L表阿霉素与2μmol/LNVP-BEZ235单独及联合作用Jurkat细胞24、48h的PI3K/Akt/mTOR信号通路中Akt、mTOR、p70s6k等表达变化。结果表阿霉素能够抑制Jurkat细胞增殖并诱导其凋亡,且凋亡作用呈浓度依赖性,5μmol/L表阿霉素作用Jurkat细胞48h的凋亡率为57.72%。在表阿霉素诱导Jurkat细胞凋亡过程中伴随Akt、mTOR、p70s6k的表达变化,NVP-BEZ235能够降低Jurkat细胞Akt、p70s6k的磷酸化水平,显著提高表阿霉素诱导Jurkat细胞凋亡的作用,5μmol/L表阿霉素和2μmol/L NVP-BEZ235联合作用Jurkat细胞48h的凋亡率达78.31%,明显高于5μmol/L表阿霉素的57.72%。结论表阿霉素抑制Jurkat细胞增殖和诱导凋亡与PI3K/Akt/mTOR信号通路有关,当该通路抑制剂与表阿霉素联用时,Jurkat细胞对于表阿霉素敏感性有一定程度的提高。
Objective To investigate the role of PI3K/Akt/mTOR signaling pathway in epirubicin-induced apoptosis and an- ti-proliferation of human T-cell lymphoma cell line Jurkat. Methods The effects of 0, 1.25, 2. 5, 5, 10μmol/L epirubiein and 0, 0. 25, 0. 5, 1,2μmol/L dual PI3K/mTOR inhibitor（ NVP-BEZ235 ） on human T-cell lymphoma cell line Jurkat proliferation after 48h was assessed by CCKS. The apoptosis of Jurkat cells with 0, 1.25, 2. 5, 5, 101μmol/L epirubicin after 48h,5μmol/L epirubicin and 2μmol/L NVP-BEZ235 after 0, 12, 24, 36 and 48h was detected by Annexin V/PE double staining flow cytometry. The effects of 5, 10μmol/L epirubicin at 0, 6, 12, 24, 48h and the change of combining 2μmol/L NVP-BEZ235 at 24, 48h with 51xmol/L epirubicin on the expressions of Akt, p-Akt, mTOR, p-mTOR, p70s6k and p-p70s6k were detected by Western blotting method. Results Epi- rubicin could inhibit proliferation of Jurkat cells and induce its apoptosis. When Jurkat was treated by 5 μmol/L epirubiein, the apopto- sis rate was 57. 72%. The apoptotic effect was concentration-dependent. Epirubicin-induced apoptosis of Jurkat cells was along with the changes of Akt, mTOR, p70s6k and their phosphorylation levels. NVP-BEZ235 reduced the phosphorylation levels of Akt and p70s6k in Jurkat cells, which significantly improved the apoptosis of Jurkat cells. The apoptotic rate rose from 57.72% to 78.31% be- cause of 2μ moL/L NVP-BEZ235 combining with 5μmol/L epirubicin after 48h. Conclusion Epirubicin-induced apoptosis of Jurkat cells has a relation with PI3 K/Akt/mTOR signaling pathway, when the pathway inhibitors combined with epirubiein, the cells sensitivi- ty of epirubicin has improved to some extent.
出处
《临床肿瘤学杂志》
CAS
2012年第9期780-784,共5页
Chinese Clinical Oncology
