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莪术油对鼻咽癌CNE22 细胞的凋亡及放疗 增敏机制的影响

Effect of Zedoary Oil for Apoptosis and Radiotherapy in Human Nasopharyngeal Carcino2 ma Cell Line CNE22

  • 摘要: 目的 探讨莪术油、莪术油联合γ2射线照射对鼻咽低分化鳞癌细胞株CNE22 的增殖抑制和诱导凋亡作用;分析莪术油是否具有放疗增敏的作用,并研究其机制。方法 体外培养CNE22 细胞,莪术油以体外直接加药的方式作用于CNE22 细胞,采用MTT、流式细胞术、电镜等分子生物学手段,检测莪术油单用或联合γ2射线对CNE22 的诱导凋亡作用、放疗增敏作用。结果 莪术油单独使用,对CNE22 细胞有确切的增殖抑制及诱导凋亡作用,并与常规化疗药物52Fu 有相近的抑制作用( P > 0. 05) 。莪术油的药物抑制作用呈现浓度依赖性,最低起效浓度为10μg/ ml, 最佳作用时间为48 小时; 5μg/ ml 、100 μg/ ml 、300μg/ ml 的莪术油联合100 cGy 的γ2射线作用2 天后凋亡率由0. 5 %、2. 15 %、10. 2 %显著提高到8. 6 %、14 %和26 %, 细胞的死亡率均在5 %以下有明显的协同增效作用( P < 0. 01) ;联合作用96h 后试验组的细胞主要以死亡为主,凋亡较少;500μg/ ml 的莪术油主要导致细胞死亡,有明显的细胞毒性。流式细胞仪结果显示较高浓度时,莪术油使CNE22 细胞阻滞在G1 期。电子显微镜观察细胞凋亡的超微结构可以看到凋亡小体。结论 莪术油可以抑制鼻咽癌细胞CNE22 的增殖并诱导其凋亡;莪术油和100 cGy 的γ2射线联合作用有明显的增敏作用; 其作用机制与诱导肿瘤细胞发生凋亡及影响细胞生长周期有关。

     

    Abstract: Objective  To explore the Zedoary oil, Zedoary oil jointγ2ray irradiation on human nasopharyn2 geal carcinoma (NPC) cell lines CNE22 inhibiting the proliferation and induction of apoptosis ; analysis of whether the Zedoary oil enhancement of the role of radiotherapy. Methods  MTT colorimet ric assay and elect ronic microscope method were employed to examine the growth status and apoptosis of CNE22 cells. Flow cytomet ry was employed to detect the effect of Zedoary oil 、Zedoary oil and/ or γ2radiation for growth inhibition in CNE22 cells. Results  Zedoary oil used alone, the CNE22 cell proliferation is the exact effect, and all with conventional chemotherapy drug 52Fu a similar effect ( P > 0. 05) ;Zedoary oil drugs inhibit a concent ration2dependent, the minimum effect f rom the concent ration of 10μg/ ml, the best time of 48 hours ; flow cytomet ry showed that 10μg/ ml, 100μg/ ml, 300μg/ ml Zedoary oil2t reated CNE22 cells the apoptosis ratio of were 0. 5 %, 2. 15 %, 10. 2 % on 48 hours, respectively. In the group of t reatment with 10μg/ ml, 100μg/ ml, 300μg/ ml curcuma and 100cGyγ2radiation, the apoptosis ratio were 8. 6 %, 14 %, 26 % on 48 hours, respectively. And the average mortality rate of cells were below 5. 0 %. They has the synergistic action for apoptosis in CNE22 cells ( P < 0. 01) . Combined 96 h af ter the test group of cells to death based, less apoptosis ;cell death was induced by 500μg/ ml curcuma. Zedoary oil is the CNE22 cell block in G1 phase. Elect ron microscopy apoptosis ult rast ructure can see that zedoary oil can be induced by CNE22 cells apoptotic body, leading to apoptosis. Conclusion  Zedoary oil to increase the in vit ro method role in CNE22 NPC cells can inhibit cell proliferation and induced apoptosis ; Zedoary oil and small doses ofγ2ray joint role obviously synergy ; mechanism and it s role in apoptosis induced by tumor cells and af2 fect cell growth cycle.

     

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