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目的:研究柯里拉京(Corilagiln)对人喉癌细胞Hep-2的抑制生长作用及凋亡诱导作用。方法 :四亚基噻唑蓝(MTT)法测定柯里拉京对细胞生长的抑制作用,流式细胞仪技术(FCM)观察细胞周期及凋亡;免疫印记法(Western-blot)测定Bax及bcl-2表达,荧光定量PCR法测量人喉癌Hep-2细胞增殖诱导配体(aproliferation-inducing ligand,APRIL)的表达量。结果:2.5～20μg/ml的柯里拉京均能抑制人喉癌细胞Hep-2生长,并有明显的时间和剂量依赖性,2.0μg/ml的柯里拉京作用72h后对人喉癌细胞Hep-2生长抑制作用达到73%;流式细胞仪检测不同浓度柯里拉京作用72h后的细胞凋亡率分别为13.80%±2.32%,21.72%±1.51%,37.43%±1.06%和54.91%±2.40%;经柯里拉京作用后,免疫印记法显示柯里拉京上调Bax,同时降低bcl-2表达,使Bax/bcl-2比值增加;柯里拉京作用人喉癌Hep-2细胞后其增殖诱导配体的mRNA表达量随时间的延长和浓度的增加而升高,48 h后各浓度与空白组比较差异有统计学意义(P<0.05)。结论:柯里拉京在体外能明显抑制人喉癌细胞Hep-2生长,抑制其生长的机制可能是诱导细胞凋亡。
Objective: To study the inhibitory effect of Corilagiln on human laryngeal carcinoma cell line Hep-2 and the induction of apoptosis. METHODS: Tetra-subunit thiazolyl blue (MTT) method was used to determine the inhibitory effect of corilagin on cell growth. Cell cycle and apoptosis were observed by flow cytometry (FCM). Bax and bcl- 2 expression was detected by fluorescence quantitative polymerase chain reaction. The expression of APRIL in human laryngeal carcinoma cell line Hep-2 was measured by fluorescence quantitative PCR. Results: Corilagin in 2.5 ~ 20μg / ml could inhibit the growth of human laryngeal carcinoma Hep-2 cells in a dose-and time-dependent manner. After treated with 2.0μg / ml corilagin for 72 hours, -2 was 73%. The apoptotic rates of cells treated with different concentrations of corilagin for 72h by flow cytometry were 13.80% ± 2.32%, 21.72% ± 1.51%, 37.43% ± 1.06% and 54.91% ± 2.40% respectively. The results of immunofluorescence showed that corilagin up-regulated Bax and decreased the expression of bcl-2, and then increased the ratio of Bax / bcl-2. The effect of cilalagin on human laryngeal carcinoma Hep-2 cell proliferation The mRNA expression of the induced ligand increased with time and concentration, and there was significant difference between each concentration and the blank group after 48 h (P <0.05). CONCLUSION: Corilagin can inhibit the growth of human laryngeal carcinoma Hep-2 cells in vitro and induce apoptosis.