论文部分内容阅读
目的:研究三七姜挥发油对人鼻咽癌(CNE-2)细胞周期的调控作用,并探讨其可能的机制。方法:体外培养人鼻咽癌细胞株(CNE-2),采用MTT法观察三七姜挥发油对CNE-2细胞增殖的抑制作用;流式细胞术分析三七姜挥发油处理CNE-2细胞后对细胞周期分布的影响;实时荧光定量PCR检测细胞周期调控相关基因的表达水平。结果:不同浓度的三七姜挥发油作用CNE-2细胞24,48,72,96 h后,均能明显抑制细胞增殖(P<0.05),且呈浓度与时间依赖性,其IC50分别为71.12,60.034,53.167,34.75μg·ml-1;三七姜挥发油诱导CNE-2细胞发生G1期阻滞,与阴性对照相组比,200μg·ml-1加药处理CNE-2细胞72 h后G1期细胞增加了13.94%(P<0.01);伴随ATM,CCND1/E1,CDK6,CDC25A,E2F1等mRNA表达水平下调,而Tp53基因表达显著增强(P<0.01)。结论:三七姜挥发油诱导人鼻咽癌CNE-2细胞G1期阻滞、抑制细胞增殖,其作用可能与抑制了介导DNA损伤修复的ATM-Chk2-Cdc25A通路,以及激活p53通路有关,从而抑制CDC25A,cyclinD1/E1、CDK6、E2F1等基因的表达而实现的。
Objective: To study the regulation effect of volatile oil of Panax Ginseng on the cell cycle of human nasopharyngeal carcinoma (CNE-2) and to explore its possible mechanism. Methods: The human nasopharyngeal carcinoma cell line (CNE-2) was cultured in vitro. MTT assay was used to observe the inhibitory effect of Radix Notoginseng volatile oil on the proliferation of CNE-2 cells. Flow cytometry Cell cycle distribution; real-time fluorescent quantitative PCR detection of cell cycle regulation related gene expression levels. Results: CNE-2 cells were significantly inhibited after 24,48,72,96 h (P <0.05) with concentration-dependent and time-dependent IC50 of 71.12, 60.034,53.167,34.75μg · ml-1; CNE-2 cells induced by the volatile oil of Panax notoginseng had G1 phase arrest. Compared with negative control group, CNE-2 cells were treated with 200μg · ml-1 for 72 h, (P <0.01). The mRNA expression of CCND1 / E1, CDK6, CDC25A and E2F1 was down-regulated while the expression of Tp53 gene was significantly increased (P <0.01). CONCLUSION: The volatile oil of Panax notoginseng can induce G1 phase arrest and inhibit the proliferation of human nasopharyngeal carcinoma CNE-2 cells, which may be related to the inhibition of the ATM-Chk2-Cdc25A pathway that mediates the DNA damage repair and the activation of p53 pathway. Inhibit the expression of CDC25A, cyclinD1 / E1, CDK6, E2F1 and other genes.