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目的探讨不同剂量柚皮素(naringenin)对人鼻咽癌CNE2细胞生长的影响及其相关作用机制。方法以不同剂量(0、5、10、20、40、60、80、100、200、400、800μg·mL~(-1))柚皮素处理CNE2细胞,作用不同时间(24、48、72 h)后,四甲基偶氮唑蓝(MTT)法检测细胞生长率的变化;以较高剂量(0、200、400μg·mL~(-1))柚皮素处理CNE2细胞48 h,进行Hochest33258荧光染色观察细胞凋亡形态变化及细胞周期流式检测;以不同剂量(0,40,200μg·mL~(-1))柚皮素处理CNE2细胞,作用不同时间(24、48 h)后,通过流式细胞仪对其活性氧含量(R0S)进行测定,并对其增殖凋亡调控相关基因P53及C-fos进行mRNA表达水平的检测。结果不同剂量柚皮素对CNE2细胞的生长具有双向调节作用,较低剂量能使CNE2细胞内活性氧含量低水平升高,从而激活细胞增殖信号途径,通过诱导C-fos(p<0.01)及抑制P53(P<0.01)基因的表达体现为促进癌细胞增殖的活性;较高剂量能引发CNE2细胞内活性氧大量产生,从而激活细胞凋亡信号途径,通过诱导P53(P<0.01)及抑制C-fos(P<0.01)基因的表达,对细胞周期产生阻滞(p<0.01)并诱导细胞凋亡,有效抑制癌细胞的增殖。结论不同剂量柚皮素对CNE2细胞生长具有双向调节作用,较低剂量体现为促癌活性,较高剂量则体现为强烈的抑癌活性。
Objective To investigate the effect of different doses of naringenin on the growth of human nasopharyngeal carcinoma cell line CNE2 and its related mechanism. Methods CNE2 cells were treated with different concentrations of naringenin (0, 5, 10, 20, 40, 60, 80, 100, 200, 400 and 800 μg · mL -1) for 24 h h), MTT assay was used to detect the cell growth rate. CNE2 cells were treated with naringenin at higher doses (0, 200, 400 μg · mL -1) for 48 h, Hoechst 33258 staining was used to observe the morphological changes and cell cycle flow cytometry. CNE2 cells were treated with different concentrations of naringenin (0, 40, 200μg · mL -1) for 24,48 h, The content of reactive oxygen species (ROS) was measured by flow cytometry, and the mRNA expression level of P53 and C-fos in proliferation and apoptosis were detected. Results Naringenin had a bidirectional effect on the growth of CNE2 cells. Lower dose of naringenin could increase the level of reactive oxygen species (ROS) in CNE2 cells and activate the cell proliferation signal pathways. By inducing C-fos (p <0.01) and The inhibition of the expression of P53 gene (P <0.01) was shown to promote the proliferation of cancer cells. Higher dose could induce a large number of reactive oxygen species production in CNE2 cells and activate the apoptotic signal pathway. By inducing P53 (P <0.01) and inhibiting C-fos (P <0.01), which could block the cell cycle (p <0.01) and induce apoptosis, effectively inhibiting the proliferation of cancer cells. Conclusion Naringenin has a bidirectional effect on the growth of CNE2 cells at different doses. The lower doses show the promotion of cancer activity, while the higher dose of naringenin shows strong anti-cancer activity.