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目的探讨小白菊内酯(parthenohde,VTL)对多发性骨髓瘤(multiple myeloma,MM)细胞的体外作用及其机制。方法培养人MM细胞系PRMI8266,在不同浓度的PTL作用不同时间后,以四甲基偶氮唑盐比色法(MTT法)及台盼蓝拒染检测细胞增殖及活性,用AO／EB染色荧光显微镜及Wright-Giemsa染色观察细胞形态学改变,应用膜联蛋白V(Annexin V)和碘化丙啶(PI)染色流式细胞仪检测细胞凋亡率,酶底物法测定细胞半胱氨酸蛋白水解酶3(Caspase-3)活性。结果(1)1- 10μmol／L的PTL对MM细胞生长曲线和活性有明显抑制作用,并呈时间和剂量依赖性,5μmol／L作用48 h,可达到接近50％的活性抑制;(2)5μmol／L PTL作用48 h后可见到细胞出现明显形态学改变,细胞形态变小、胞质减少、胞体胞核固缩、出现凋亡小体;(3)2μmol／L、5μmol／L、10μmol／L的PTL作用48 h后MM细胞的凋亡率分别为17．1％±2．6％、33．6％±3．8％、40．9％±3．1％,与对照组5．6％±1．2％相比,差异有显著统计学意义(均P<0．01);(4)PTL作用48 h后,MM细胞caspase-3活性明显增强,且呈浓度依赖性(P<0．01)。结论。PTL能明显抑制MM细胞体外生长及活性,其作用机制为增强caspase-3活性而诱导MM细胞凋亡;PTL作为新的MM细胞凋亡诱导剂,其作用靶点及体内效应值得深入研究。
Objective To investigate the in vitro effect and mechanism of Parthenohde (VTL) on multiple myeloma (MM) cells. Methods The human MM cell line PRMI8266 was cultured and treated with different concentrations of PTL for different time. The cell proliferation and activity were detected by MTT assay and trypan blue exclusion. AO / EB staining Fluorescence microscopy and Wright-Giemsa staining were used to observe the morphological changes of the cells. The apoptotic rate was detected by flow cytometry with Annexin V and propidium iodide (PI) staining. Cysteamine Acid protease 3 (Caspase-3) activity. Results (1) PTL of 1 ~ 10μmol / L significantly inhibited the growth curve and activity of MM cells in a dose- and time-dependent manner. At 5μmol / L for 48 h, the inhibitory activity of PTL reached nearly 50% After treated with 5μmol / L PTL for 48h, obvious morphological changes of the cells were observed, the morphology of the cells was diminished, the cytoplasm was reduced, and the apoptotic bodies were observed. (3) The apoptotic bodies were observed at the concentration of 2μmol / L, 5μmol / L, 10μmol / L for 48 h, the apoptosis rates of MM cells were 17.1% ± 2.6%, 33.6% ± 3.8%, 40.9% ± 3.1% (P <0.01). (4) The activity of caspase-3 in MM cells was significantly increased in a dose-dependent manner (P <0.01) P <0.01). in conclusion. PTL can significantly inhibit the growth and activity of MM cells in vitro, and its mechanism of action is to increase the activity of caspase-3 and induce the apoptosis of MM cells. PTL as a new apoptosis inducer of MM cells, its target of action and in vivo effect worth further study.