摘要
探讨薏苡仁酯（coixenolide,CXL）对同源不同辐射抗性鼻咽癌细胞增殖及凋亡的影响,探讨其可能的作用机制。采用CCK-8法、DAPI染色法及流式细胞术检测CXL作用后CNE-2R和CNE-2细胞活性及凋亡的情况,Western blotting法检测凋亡相关基因的蛋白表达水平。CCK-8法显示CXL呈剂量-时间依赖性的抑制CNE-2R（24 h,48 h,72 h的IC_（50）分别为273.20 g/L,10.88 g/L和4.55 g/L）和CNE-2（24 h,48 h,72 h的IC_（50）分别为256.60 g/L,5.44 g/L和2.43 g/L）的细胞活性;DAPI染色显示CXL组细胞中观察到染色质浓缩、边集和分割成块状;与对照组相比,流式细胞术显示CXL作用于CNE-2R（（4.51依0.09）vs（11.68±3.54）,p〈0.05）和CNE-2（（5.27±0.14）vs（15.68±2.76）,p〈0.05）细胞凋亡率均明显增加;Western blotting法显示,CXL组Bax、Caspase-9、cleaved-caspase-3、cleaved-PARP蛋白表达量较对照组均显著增加（p〈0.05）,Bcl-2蛋白表达量显著降低（p〈0.05）,Bcl-2/Bax比值降低（p〈0.001）。上述均表明CXL可抑制同源不同辐射抗性鼻咽癌细胞的增殖及诱导其凋亡,其作用机制可能与凋亡调节基因的表达变化有关。
To investigate the effects of coixenolideon on proliferation and apoptosis in homologous nasopharyngeal arcinoma cells with different radioresistance,and to explore the possible mechanism.CCK-8 assay,DAPI staining and flow cytometry with Annexin V/PI staining were applied to detect viability and apoptosis of CNE-2 R and CNE2 cells after treated with CXL,respectively.The protein expression level of several genes related to apoptosis was detected by Western blotting.CCK-8 assay showed that CXL produced an obvious dose-time dependent inhibitory effect on the viability of CNE-2 R cells with the IC_（50） of 273.20 g/L、10.88 g/L、4.55 g/L and CNE-2 cells with the IC_（50） of 256.60 g/L、5.44 g/L、2.43 g/L for 24 h、48 h、72 h respectively.DAPI staining showed that chromatin condensation,margination and splitting into a massive structure were observed in CXL cells array.Flow cytometry with Annexin V/PI staining showed that the apoptotic rate of CNE-2 R cells was apparentlyincreased（P〈0.05）from（4.51±0.09）% to（11.68±3.54）% compareed to CNE-2 cells which was significantly increased（P〈0.05） from（5.27±0.14）% to（15.68±2.76）%.Western blotting showed that after treated with CXL,the expression of PARP、cleave-Caspase-3、Caspase-9、Bax protein significantly increased（P〈0.05） corresponding to the control groups.The expression of Bcl-2 protein significantly decreased（P〈0.05）.The protein expression ratio of the Bcl-2 to Bax decreased（P〈0.05）.The results indicated that CXL could inhibit the proliferation and induce the apoptosis of the homologous nasopharyngeal arcinoma cells with different radioresistance.It is possibly mediated by the varyingexpression ofthe apoptosis-regulatorygenes.
作者
刘静姝
周寒静
隆姝孜
刘洪良
杨扬
张涛
Liu Jingshu;Zhou Hanjing;Long Shuzi;Liu Hongliang;Yang Yang;Zhang Tao(The First Affiliated Hospital of Chongqing Medical University,Chongqing,400016)
出处
《基因组学与应用生物学》
CAS
CSCD
北大核心
2018年第8期3587-3592,共6页
Genomics and Applied Biology
基金
重庆市卫生和计生委中医药科技项目(ZY20150245)
肿瘤科国家临床重点专科建设项目(国卫办医函(2013)544号)共同资助
关键词
鼻咽癌
薏苡仁酯
辐射抗拒
增殖
凋亡
Nasopharyngeal carcinoma
Coixenolide
Radioresistance
Proliferation
Apoptosis
