Proteomic Analysis of Normal and Cancer Cervical Cell Lines Reveals Deregulation of Cytoskeleton-associated Proteins
Proteomic Analysis of Normal and Cancer Cervical Cell Lines Reveals Deregulation of Cytoskeleton-associated Proteins
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DOI：
10.21873/cgp.20036
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发表时间：
2017-07-01
影响因子：
2.5
通讯作者：
Anagnou, Nicholas P.
中科院分区：
文献类型：
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作者：
Pappa, Kalliopi I.;Lygirou, Vasiliki;Anagnou, Nicholas P.
背景：HPV阳性和阴性宫颈癌主要与细胞周期和细胞骨架紊乱有关；然而，影响的实际生物学过程仍然难以捉摸。为此，我们系统地描述了四个不同的和信息丰富的宫颈细胞系的细胞内蛋白质组特征。材料和方法：对正常宫颈(HCK1T)、HPV阴性(C33A)、HPV阳性(HPV16+)和HeLa(HPV18+)三种宫颈癌细胞株的细胞提取液进行双向电泳分析，MALDI-TOF质谱仪鉴定差异表达蛋白质，Western印迹分析鉴定差异表达蛋白质。结果：共发现133个蛋白在正常宫颈组织和宫颈癌细胞系中存在差异表达。生物信息学分析表明，肌动蛋白细胞骨架信号通路受到显著影响，而肌动蛋白解聚因子cofilin-1的上调被证实，并通过Western blotting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个信息性差异表达的蛋白质。结论：这些新鉴定的蛋白质为进一步的功能研究提供了动力，以剖析宫颈癌发生的两条不同途径的机制。
Background: 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. Materials and Methods: 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. Results: In total, 133 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. Conclusion: These novel identified proteins provide the impetus for further functional studies to dissect the mechanisms operating in the two distinct pathways of cervical carcinogenesis.