Pappa Kalliopi I, Lygirou Vasiliki, Kontostathi Georgia, Zoidakis Jerome, Makridakis Manousos, Vougas Konstantinos, Daskalakis George, Polyzos Alexander, Anagnou Nicholas P
First Department of Obstetrics and Gynecology, University of Athens School of Medicine, Alexandra Hospital, Athens, Greece
Cell and Gene Therapy Laboratory, Centre of Basic Research II, Biomedical Research Foundation of the Academy of Athens (BRFAA), Athens, Greece.
Cancer Genomics Proteomics. 2017 Jul-Aug;14(4):253-266. doi: 10.21873/cgp.20036.
Both HPV-positive and -negative cervical cancers are primarily associated with features of cell cycle and cytoskeletal disruption; however, the actual biological processes affected remain elusive. To this end, we systematically characterized the intracellular proteomic profiles of four distinct and informative cervical cell lines.
Cell extracts from a normal cervical (HCK1T) and three cervical cancer cell lines, one HPV-negative (C33A), and two HPV-positive, SiHa (HPV16+) and HeLa (HPV18+), were analyzed by 2-dimensional electrophoresis and differentially expressed proteins were identified by MALDI-TOF mass spectrometry, while differential expression was confirmed by western blot analysis.
In total, 113 proteins were found differentially expressed between the normal and the cervical cancer lines. Bioinformatics analysis revealed the actin cytoskeleton signaling pathway to be significantly affected, while up-regulation of cofilin-1, an actin depolymerizing factor, was documented and further validated by western blotting. Furthermore, two-way comparisons among the four cell lines, revealed a set of 18 informative differentially expressed proteins.
These novel identified proteins provide the impetus for further functional studies to dissect the mechanisms operating in the two distinct pathways of cervical carcinogenesis.
HPV阳性和阴性宫颈癌均主要与细胞周期和细胞骨架破坏的特征相关;然而,受影响的实际生物学过程仍不清楚。为此,我们系统地表征了四种不同且信息丰富的宫颈细胞系的细胞内蛋白质组图谱。
通过二维电泳分析来自正常宫颈(HCK1T)和三种宫颈癌细胞系(一种HPV阴性(C33A)以及两种HPV阳性,SiHa(HPV16+)和HeLa(HPV18+))的细胞提取物,并用基质辅助激光解吸电离飞行时间质谱法鉴定差异表达的蛋白质,同时通过蛋白质印迹分析确认差异表达。
总共发现113种蛋白质在正常细胞系和宫颈癌细胞系之间差异表达。生物信息学分析显示肌动蛋白细胞骨架信号通路受到显著影响,同时记录了肌动蛋白解聚因子cofilin-1的上调,并通过蛋白质印迹进一步验证。此外,四种细胞系之间的双向比较揭示了一组18种信息丰富的差异表达蛋白质。
这些新鉴定的蛋白质为进一步的功能研究提供了动力,以剖析在宫颈癌发生的两种不同途径中起作用的机制。