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目的:研究白花蛇舌草多糖(PEHD)体外抑制鼻咽癌CNE2细胞株增殖,诱导细胞凋亡及其凋亡机制。方法:用不同剂量PEHD(2、4、6mg/ml)处理CNE2细胞24h、48h、72h,用四甲基偶氮唑蓝法(MTT)检测CNE2细胞的增殖情况,计算抑制率。在不同药物浓度(2、4、6mg/ml)PEHD作用48h后,用Annexin V-FITC/碘化丙锭双染法(Annexin V/PI)标记的流式细胞术检测CNE2细胞的凋亡率。采用Western blot检测用药前后细胞中Bax蛋白、Bcl-2蛋白和caspase-3蛋白的表达水平。结果:MTT结果显示,2、4、6mg/ml PEHD可以明显抑制CNE2细胞增殖(均P<0.05),最高抑制率可达76.5%,在2~6mg/ml浓度内随着浓度的增加、时间的延长抑制作用逐渐增强,呈时间-剂量依赖性。流式细胞术检测到4、6mg/ml PEHD作用48h后,CNE2凋亡细胞比例显著增加,凋亡率分别为31.32%、46.28%,高于空白对照组4.86%(P<0.01)。Western blot显示PEHD处理48h后,CNE2细胞Bax蛋白和caspase-3蛋白表达上升,Bcl-2的表达下降。结论:在一定浓度范围内PEHD(2、4、6mg/ml)能够明显抑制鼻咽癌CNE2细胞的增殖,呈时间-剂量依赖性,其抑制作用与诱导细胞凋亡有关;PEHD可通过上调Bax、caspase-3蛋白表达、下调Bcl-2蛋白表达,诱导CNE2细胞凋亡。
AIM: To investigate the inhibitory effect of Hedyotis diffusa Willd polysaccharides (PEHD) on CNE2 nasopharyngeal carcinoma cell proliferation, apoptosis and apoptosis in vitro. Methods: CNE2 cells were treated with different doses of PEHD (2,4,6mg / ml) for 24h, 48h, 72h. The proliferation of CNE2 cells was detected by MTT assay and the inhibition rate was calculated. After treated with PEHD for different concentrations (2,4 and 6 mg / ml) for 48 h, the apoptotic rate of CNE2 cells was detected by flow cytometry with Annexin V-FITC / Annexin V / PI staining . Western blot was used to detect the expression of Bax protein, Bcl-2 protein and caspase-3 protein in the cells before and after treatment. Results: MTT results showed that 2,4,6mg / ml PEHD could significantly inhibit the proliferation of CNE2 cells (all P <0.05), the highest inhibition rate was 76.5%. With the increase of concentration of 2 ~ 6mg / ml, Prolong the inhibitory effect gradually increased in a time-dose-dependent manner. Flow cytometry showed that the proportion of apoptotic cells in CNE2 significantly increased after 4 and 6mg / ml PEHD treatment for 48h, respectively. The apoptotic rate was 31.32% and 46.28%, respectively, which was higher than that of blank control group (4.86%, P <0.01). Western blot analysis showed that the expression of Bax protein and caspase-3 protein in CNE2 cells decreased and the expression of Bcl-2 decreased after 48 h PEHD treatment. CONCLUSION: PEHD (2, 4, 6 mg / ml) can significantly inhibit the proliferation of CNE2 cells in a dose-and time-dependent manner in a certain concentration range, and its inhibitory effect is related to the induction of apoptosis. PEHD can up- , Caspase-3 protein expression, down-regulate Bcl-2 protein expression and induce CNE2 cell apoptosis.