Mechanism of apoptosis of nasopharyngeal carcinoma cells induced by polysaccharides extracts from Hedyotic diffusa
-
摘要: 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、6 mg/ml)处理CNE2细胞24 h、48 h、72 h,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、6 mg/ml)PEHD作用48 h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、6 mg/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2~6 mg/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、6 mg/ml PEHD作用48 h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48 h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、6 mg/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。Abstract: Objective:To explore the proliferation inhibition and apoptosis of polysaccharides extracts from polysaccharides extracts from Hedyotic diffusa (PEHD) on Human Nasopharyngeal Carcinoma (NPC)cell line CNE2 cells in vitro.Method:CNE2 cells treated with various concentrations of PEHD were detected by MTT assay at 24 h, 48 h, and 72 h. The apoptotic cells were analyzed by flow cytometry with Annexin V/PI staining. The expression levels of Bax, Bcl-2 and caspase-3 protein were examined by Western blotting method.Result:The growth of CNE2 cells were suppressed after treatment with PEHD (P<0.05), MTT assay showed that the highest cell inhibition rate reached to 76.5%, the inhibition in the doses from 2 to 6 mg/ml showed dose-and-time-dependent. The percent of apoptosis in 4 and 6 mg/ml PEHD treatment groups for 48 h were 31.32%, 46.28%,respectively, and significantly higher than that in control groups, 4.86% (P<0.01).After the cells being treated with PEHD for 48 h, the expression of Bax and caspase-3 protein increased, and the expression of Bcl-2 protein decreased gradually.Conclusion:PEHD could inhibited the growth of CNE2 cells and was dose-and-time-dependent, the mechanism may involve induction of cell apoptosis, which was associated with the activation of Bax and caspase-3 protein and the down-regulation of Bcl-2 protein expression.
-
-
[1] AGULNIK M,EPSTEIN J B.Nasopharyngeal carcinoma:current management,future directions and dental implications[J].Oral Oncol,2008,44:617-627.
[2] 韦星,万福生,涂硕,等.白花蛇舌草注射液诱导人肺癌细胞株SPC-A-1凋亡及其分子机制的研究[J].中国老年学杂志,2007,27(2):110-112.
[3] 孟庆宇,吕秀芳,任凤云.白花蛇舌草多糖对Bel7402细胞基因表达的影响[J].中国老年学杂志,2008,28(13):1271-1272.
[4] 蒋剑平,许海顺,卢烨琳,等.响应面分析法优化白花蛇舌草水溶性多糖的提取工艺[J].浙江中医药大学学报,2012,36(2):187-191.
[5] 赵浩如,李瑞,林以宁,等.白花蛇舌草不同提取工艺对抗肿瘤活性的影响[J].中国药科大学学报,2002,33(6):510-513.
[6] LING Y Z.Separation purify and content determination of polysassharides in hedyotis diffusa willd[J].Biotechnology,2005,15:48-50.
[7] ANDERSSON D,LIU J J,NILSSON A,et al.Ursolic acid inhibits proliferation and stimulates apoptosis in HT29 cells following activation of alkaline sphingomyelinase[J].Anticancer Res,2003,24:3317-3322.
[8] 曾永长,梁少瑜,罗佳波,等.白花蛇舌草提取物体内外抗肿瘤实验研究[J].中药材,2011,34(4):594-597.
[9] 赵晓艳,胡玉娜,康向东,等.熊果酸诱导人胃癌BGC823细胞凋亡及其作用机制初探[J].中国癌症杂志,2010,20(2):101-104.
[10] PLATI J,BUCUR O,KHOSRAVI-FAR R.Apoptotic cell signaling in cancer progression and therapy[J].Integra Biol(Camb),2011,3:279-296.
[11] 林圣云,沈楚云,蒋剑平,等.白花蛇舌草多糖诱导多发性骨髓瘤细胞凋亡及其机制研究[J].中华血液学杂志,2013,34(4):337-340.
[12] THORNBERRY N A,LAZEBNIK Y.Caspases:enemies within[J].Science,1998,281:1312-1316.
[13] CORY S,ADAMS J M.Killing cancer cells by flipping the Bcl-2/Bax switch[J].Cancer Cell,2005,8:5-6.
[14] TULALAMBA W,JANVILISRI T.Nasopharyngeal carcinoma signaling pathway:an update on molecular biomarkers.[J].Int J Cell Biol,2012,5:594681.
-
计量
- 文章访问数: 373
- PDF下载数: 122
- 施引文献: 0
下载: