摘要目的 探讨雷公藤甲素(TP)对急性髓系白血病伴FMS样酪氨酸激酶3内部串联重复(FLT3-ITD)的细胞株MV411增殖和凋亡的影响,及其对PI3K-Akt-mTOR通路的作用.方法 四甲基偶氮唑盐(MTT)法测定不同浓度TP作用24、48、72 h时MV411细胞的增殖抑制率;流式细胞术检测48、72 h的细胞凋亡率;实时荧光定量聚合酶链反应(PCR)检测PI3K-Akt-mTOR通路相关基因FLT3、PTEN、PI3K、Akt、mTOR mRNA的表达.结果0、5、10、20、40、80 nmol/L TP作用24、48、72 h,MV411细胞增殖受到抑制,呈现时间-剂量依赖性.0、10、20 nmol/L TP作用48 h后,MV411细胞平均早期凋亡率分别为(3.30±0.20)%、(17.10±0.36)%、(35.67±0.61)%,作用72 h后分别为(7.37±0.32)%、(49.33±0.40)%、(68.92±0.11)%,同一时间各浓度组间差异均有统计学意义(均P=0.000).TP能明显降低FLT3、PI3K、Akt、mTOR mRNA的表达,升高PTEN mRNA的表达.结论TP可能通过影响PI3K-Akt-mTOR通路相关基因的表达发挥抑制MV411细胞增殖、诱导其凋亡的作用.

abstractsObjective To investigate the effects of triptolide (TP) on proliferation and apoptosis of acute myeloid leukemia cell line MV411 with FMS-like tyrosine kinase 3 internal tandem duplication (FLT3-ITD), and its effect on PI3K-Akt-mTOR pathway. Methods MTT assay was used to detect the proliferation inhibition of MV411 cell proliferation treated by different concentrations of TP in 24, 48 and 72 h. Flow cytometry was used to measure the cell apoptosis rate at 48 and 72 h. Real-time fluorescent quantitative PCR was used to detect the expression of FLT3, PTEN, PI3K, Akt, mTOR mRNA on PI3K-Akt-mTOR pathway. Results After treated by 0, 5, 10, 20, 40, and 80 nmol/L TP in 24, 48 and 72 h, the viability of MV411 cells was inhibited in a time-dose dependence manner. MV411 cells was treated by 0, 10, and 20 nmol/L TP after 48 and 72 h, the cell apoptosis rates were rising with the increasing of drug concentration, there were statistical significances [48 h:(3.30±0.20) %, (17.10±0.36) %, (35.67±0.61) %, 72 h: (7.37±0.32) %, (49.33± 0.40)%, (68.92±0.11)%, all P=0.000]. The expressions of FLT3, PI3K, Akt, and mTOR mRNA were down-regulated and PTEN mRNA expression was up-regulated by TP. Conclusion TP may inhibit the proliferation and induce apoptosis of MV411 cells by affecting the expression of PI3K-Akt-mTOR pathway related genes.