吲哚-3-原醇对人鼻咽癌CNE-2细胞增殖和凋亡的影响  

Effect of Indole-3-carbinol on Proliferation and Apoptosis of Human Nasopharyngeal Carcinoma CNE-2 Cells

作　　者：王茜[1] 杜敬东[1] 毛华东[1] 程杰[1] 

机构地区：[1]三峡大学仁和医院,湖北宜昌443001

出　　处：《广州中医药大学学报》2015年第2期331-335,共5页Journal of Guangzhou University of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(编号:81100264)

摘　　要：【目的】探讨吲哚-3-原醇(indol-3-carbinol,I3C)对人低分化鼻咽癌细胞株CNE-2的作用及其机制。【方法】将对数生长期的CNE-2细胞分成空白对照组和I3C组。空白对照组细胞常规培养,I3C组细胞培养体系中加入I3C(50μmol/L)。各组细胞培养48 h后,采用四甲基偶氮唑盐(MTT)法检测细胞增殖;Annexin V-碘化丙啶/异硫氰酸荧光素(PI/FITC)检测细胞凋亡,Real-time PCR检测细胞中胞外信号调节激酶(ERK)和Bax、Bcl-2基因表达;Western blot法检测细胞中ERK、p-ERK、Bax、Bcl-2蛋白表达,Caspase3活性试剂盒检测Caspase3活性。【结果】I3C组与空白对照组比较,CNE-2细胞增殖率显著下降(P<0.05),凋亡率显著升高(P<0.05),Real-time PCR检测显示I3C组与空白对照组比较,ERK m RNA表达无显著性差异,但Bax m RNA表达水平显著增高(P<0.001),Bcl-2 m RNA表达水平显著下降(P<0.05)。Western blot法检测显示,I3C组与空白对照组比较,ERK总蛋白无明显变化(P>0.05),但p-ERK和Bcl-2蛋白表达显著减少(P<0.05),Bax蛋白表达和Caspase3活性显著增加(P<0.05)。【结论】吲哚-3-原醇能通过抑制ERK信号传导通路激活而抑制CNE-2细胞增值和促进CNE-2细胞凋亡。Objective To investigate the effect of indole-3-carbinol（ I3C） on the human poorly differentiated nasopharyngeal carcinoma cell strain CNE-2 and to explore the possible mechanism. Methods The CNE-2 cells at logarithmic growth phase were divided into blank control group and I3 C group. The cells in the blank control group were cultured with conventional medium, and cells in I3 C group were incubated with conventional medium and I3C（50 μmol / l）. Methyl thiazolyl tetrazolium（MTT） assay was used to examine the proliferation and Annexin V-propidium iodide / fluorescein isothiocyanate（PI / FITC） was used for detecting the apoptosis in the two groups after incubation for 48 h. The m RNA levels of signal-regulated kinase（ERK）, Bax, and Bcl-2in CNE-2 cells were detected by real-time PCR, and their protein expression levels were detected by Western blotting method. The activity of Caspase3 was measured by Caspase3 colorimetric assay kit. Results Compared with the blank control group, the proliferation rate of CNE-2 cells in I3 C group was significantly decreased（P〈0.05）, but the apoptotic rate of cells was significantly increased（ P〈0.05）. After treatment with I3 C, the expression level of ERK m RNA showed no obvious difference, but the m RNA level of Bax was significantly increased（ P〈0.001） and Bcl-2 was decreased（ P〈0.05）. The Western blotting results showed total ERK protein level in I3 C group did not differ from that of the blank control group（ P〉0.05）, but the expression levels of p-ERK and Bcl-2 were significantly decreased while Bcl-2 level and Caspase 3 activity were increased（ P〈0.05）. Conclusion Indole-3-carbinol could inhibit the proliferation and promote the apoptosis of CNE-2cells through attenuating the activation of ERK signal transduction pathway.

关 键 词：吲哚-3-原醇/药理学 细胞增殖 细胞凋亡 细胞培养 

分 类 号：R285.5[医药卫生—中药学]