蛋白质组学技术在研究AdIL-24抗癌机制中的应用

Application of Proteomics Technique in Research of the Anticancer Mechanisms of AdIL-24

目的:探讨蛋白质组学技术在研究AdIL-24抗宫颈癌CaSki细胞作用机制的应用价值。方法:前期研究已成功构建AdIL-24,可抑制宫颈癌CaSki细胞生长,促进凋亡。在此基础上,通过双向凝胶电泳分离AdIL-24处理前后细胞总蛋白,并对胶条(pH3-10NL、pH4-7)、SDS-PAGE浓度(10%、12%)、电泳时间(6h、7h)进行优化,PDQuest图像分析,MALDI-TOF质谱鉴定,经Mascot、MS-fit软件查询SWISS-PORT数据库鉴定差异蛋白。结果:获得分辨率高、重复性好的CaSki细胞双向电泳图谱,质谱鉴定的差异蛋白点经Mascot、MS-fit软件搜索到同一种蛋白质。结论:已建立的蛋白质组学技术,为进一步研究AdIL-24的抑癌机制奠定基础。

Objective： To evaluate the role of proteomics technique in studying on the mechanisms of AdIL- 24 in inhibiting cervical cancer CaSki cells. Methods： The inhibitory effect of AdIL- 24 to CaSki cells and apoptosis induced were verified by early research. Total proteins of treated cells and control cells were extracted and separated by two dimensional gel electrophoresis（2 - DE） ,conditions（gel strip： pH3 - 10NL/pH4 - 7; SDS-PAGE concentration： 10%/12%; electrophoresis time： 6h/7h） were adjusted , the differential expression proteins were analyzed by PDQuest software,and identified using matrix- assisted laser desorption/ionization time of flight mass spectrometry （MALD1- TOF- MS） and Mascot,MS- fit database searching.Results： 2 - DE patterns of CaSki cells with high - resolution and reproducibihty were obtained. The same protein was obtained by Mascot and MS - fit database searching after mass spectrometry identification. Conclusion： Proteomics technique that we established can be effectively applied to study the anticancer mechanisms of AdIL - 24.