摘要
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 explore the influence and the mechanisms of different dose naringenin on nasopharyngeal carcinoma CNE2 cells growth. METHODS Different concentrations of naringenin （0,5,10,20,40,60,80,100,200,400,800 μg·mL-1 ）were used to detect their effects on CNE2 cells growth for 24 to 72 h by MTF Method. High concentrations of naringenin （0,200,400 μg·mL-1） for 48 h were used to detect apoptosis by Hochest 33 258 staining. High concentrations of naringenin （0,200,400 μg·mL-1 ） for 48 h were used to detect the cell cycle by flow cytometry. Different concentrations of naringenin （0,40,200μg·mL-1） for 24 and 48 h were used to detect the level of reactive oxygen species （ROS） by flow cytometry, and mRNA expression levels of P53 and C-los were determined by qPCR. RESULTS After different concentrations of naringenin administration, CNE2 cells were changed growth rate. Relatively low dose can induce the low level rise of ROS, the low level rise of ROS can induce the up-regulation of the ex- pression levels of c-los （P 〈 0. 01 ）and the down-regulation of the expression levels of P53 （P 〈 0. 01 ） , so relatively low dose can pro- mote CNE2 cells growth rate . However, relatively high dose can induce the high level rise of ROS, the high level rise of ROS can in- duce the up-regulation of the expression levels of P53 （ P 〈 0. 01 ） and the down-regulation of the expression levels of C-fos（ P 〈 0. 01 ）, so relatively high dose can induce apoptosis, also possess the effect of S and Gz/M phase blockage（ P 〈 0. 01 ） , significantly suppress CNE2 cells growth rate. CONCLUSION Different dose naringenin has two-way effects on CNE2 cells growth, relatively low dose has carcinogenic effect, relatively high dose has good anticancer effect.
出处
《中国药学杂志》
CAS
CSCD
北大核心
2013年第23期2006-2012,共7页
Chinese Pharmaceutical Journal
基金
广西自然科学基金资助项目(2010GXNSFA013254)
广西医学科学实验中心开放基金(KFJJ2010-57)
广西教育厅科研项目(NO.201010LX328)
关键词
柚皮素
不同剂量
CNE2
促癌
抑癌
naringenin
different dose
CNE2
carcinogenic effect
anticancer effect
