CAJ | 학술논문

目的探讨与鼻咽癌细胞放射敏感性相关的并可能预测鼻咽癌细胞放射敏感性的蛋白质.方法以人鼻咽癌细胞株CNE-2诱导建立放射抗拒的鼻咽癌细胞株CNE-2( R743),通过克隆形成实验及流式细胞术检测以比较CNE-2和CNE-2( R743)细胞的放射敏感性及细胞周期的特征.提取细胞总蛋白质,进行双向凝胶电泳,Image Master 7.0图像分析软件分析图谱以识别差异蛋白质点,MALDI-TOF/TOF肽质量指纹图谱分析,蛋白质质谱数据库搜索鉴定差异蛋白.应用RTPCR和Western blot验证2种细胞中相关差异蛋白的表达.结果得到7个差异表达较显著的蛋白质,与CNE-2相比较,CNE-2( R743)细胞中6个上调,1个下调.Western blot及RT-PCR证实膜联蛋白A2、原肌球蛋白及糖调节蛋白78在CNE-2( R743)的表达均上调(t=24.22、24.20和29.19,P＜0.05),与蛋白质组学结果一致.结论不同放射敏感性鼻咽癌细胞存在差异表达的蛋白质,其中膜联蛋白A2、原肌球蛋白及糖调节蛋白78有可能成为预测鼻咽癌细胞放射敏感性的候选生物标志物.
목적 탐토여비인암세포방사민감성상관적병가능예측비인암세포방사민감성적단백질.방법 이인비인암세포주CNE-2유도건립방사항거적비인암세포주CNE-2( R743),통과극륭형성실험급류식세포술검측이비교CNE-2화CNE-2( R743)세포적방사민감성급세포주기적특정.제취세포총단백질,진행쌍향응효전영,Image Master 7.0도상분석연건분석도보이식별차이단백질점,MALDI-TOF/TOF태질량지문도보분석,단백질질보수거고수색감정차이단백.응용RTPCR화Western blot험증2충세포중상관차이단백적표체.결과 득도7개차이표체교현저적단백질,여CNE-2상비교,CNE-2( R743)세포중6개상조,1개하조.Western blot급RT-PCR증실막련단백A2、원기구단백급당조절단백78재CNE-2( R743)적표체균상조(t=24.22、24.20화29.19,P＜0.05),여단백질조학결과일치.결론 불동방사민감성비인암세포존재차이표체적단백질,기중막련단백A2、원기구단백급당조절단백78유가능성위예측비인암세포방사민감성적후선생물표지물.
Objective To investigate the proteins which were associated with radiosensitivity of nasopharyngeal carcinoma (NPC) cells and could be used to predict the radiosensitivity.Methods A radioresistant subclone cell line CNE-2 (R743) derived from NPC cell line CNE-2 was established.Radiosensitivity and cell cycle characteristics of CNE-2 and CNE-2 ( R743 ) were examined and compared by clonogenic survival assay and flow cytometry.The total proteins from the two cell lines were extracted and separated by two-dimensional gel electrophoresis,and the images were analyzed by Image Master 7.0analysis software.Differentially expressed proteins in the two cell lines were identified through MALDITOF/TOF peptide mass fingerprint and searched in the protein sequence database.The protein expressions were confirmed by RT-PCR and Western blot.Results Totally seven differentially expressed proteins were identified,six of which were upregulated and one downregulated in the radioresistant CNE-2 (R743),compared with those of CNE-2.Three out of the seven,Annexin A2,Tropomyosin 4 and GRP78 were upregulated in the CNE-2 ( R743 ),which were confirmed by Western blot and RT-PCR ( t =24.22,24.20,29.19,P ＜ 0.05).Conclusions Differentially expressed proteins might be involved in different radiosensitivities of nasopharyngeal carcinoma cell lines,among which Annexin A2,Tropomyosin 4 and GRP78 could be the candidate biomarkers for predicting radiosensitivity of nasopharyngeal carcinoma cells.