CAJ | 학술논문

背景与目的紫杉醇是用于肺癌临床一线治疗的作用于微管蛋白的化疗药物,而耐药是影响其疗效的关键因素.本研究应用蛋白组学技术整体比较紫杉醇耐药细胞与敏感细胞分泌蛋白表达谱,以期发现肺癌紫杉醇耐药相关分泌蛋白标志物,为临床选择有针对性的化疗药物提供依据.方法收集A549亲代及耐药细胞培养液中的蛋白,应用差异凝胶电泳(DIGE)进行分离,Decyder 6.5软件分析后获得的差异蛋白点通过基质辅助激光解析电离飞行时间质谱(MALDI-TOF-MS)进行鉴定.结果应用DIGE进行分离后,成功获得了蛋白质组分辨率高、重复性好的双向凝胶电泳图谱,Decyder 6.5软件分析获得差异>2倍以上的蛋白质点共有76个,经质谱鉴定了19种蛋白,鉴定的蛋白涉及代谢酶类、细胞外基质降解酶类、粘附分子、细胞因子和信号转导相关蛋白质.结论本研究首次应用双向电泳技术整体展示了紫杉醇耐药细胞株及敏感细胞株的分泌蛋白表达谱,并分析、鉴定、筛选出与紫杉醇耐药相关的差异分泌蛋白.该研究为非小细胞肺癌血清耐药标志物的筛选提供了新的方法和候选分子.
배경여목적 자삼순시용우폐암림상일선치료적작용우미관단백적화료약물,이내약시영향기료효적관건인소.본연구응용단백조학기술정체비교자삼순내약세포여민감세포분비단백표체보,이기발현폐암자삼순내약상관분비단백표지물,위림상선택유침대성적화료약물제공의거.방법 수집A549친대급내약세포배양액중적단백,응용차이응효전영(DIGE)진행분리,Decyder 6.5연건분석후획득적차이단백점통과기질보조격광해석전리비행시간질보(MALDI-TOF-MS)진행감정.결과 응용DIGE진행분리후,성공획득료단백질조분변솔고、중복성호적쌍향응효전영도보,Decyder 6.5연건분석획득차이>2배이상적단백질점공유76개,경질보감정료19충단백,감정적단백섭급대사매류、세포외기질강해매류、점부분자、세포인자화신호전도상관단백질.결론 본연구수차응용쌍향전영기술정체전시료자삼순내약세포주급민감세포주적분비단백표체보,병분석、감정、사선출여자삼순내약상관적차이분비단백.해연구위비소세포폐암혈청내약표지물적사선제공료신적방법화후선분자.
Background and objective Paclitaxol (PTX) resistance is one of main factors which affect the outcome of chemotherapy of lung adenocarcinoma. The aim of this study is to compare the secreted protein expression profiles between Paditaxol (PTX) resistant and sensitive lung adenocarcinoma cells by proteomic research method, so as to provide evidence of choosing individual chemotherapy drugs in clinical treatment. Methods Total secreted proteins extracted from a PTX sensitive cell line A549 and a PTX resistant cell line A549-Taxol were separated by fluorscent differential gel electrophoresis (DIGE). High quality 2-DE profiles were obtained and analyzed by Decyder 6.5 analysis software to screen differentially expressed protein spots. Those spots were identified by mass spectrometry. Results 2-DE patterns of lung adenocarcinoma cells with high-reso-lution and reproducibility were obtained. 76 significantly differentially expressed protein spots were screened, 19 proteins were identified by mass spectrometry. The identified proteins could be classified into different catogories: metabolic enzyme, extracel-lular matrix (ECM) degradation enzyme, cytokine, signal transducer, cell adhesion, and so on. Conclusion Multiple secreted proteins related to chemoresistance of A549-Taxol cells were identified in this study for the first time. The results presented here would provide dues to identify new serologic chemoresistant biomarkers of NSCLC.