| Objective: Analyze different expressions of nucleus proteins andα-tubulin,β-tubulin andγ-tubulin between HPV16-E6 immortalized human squamous epithelial cell (H8) and cervical cancer cell (Caski); identify different expressions of nucleus proteins. Results from the subproteomic analysis could be useful for the understanding of the mechanism of carcinogenesis and optioning a target of anticancer drug.Methods:①The nucleus proteins and the skeleton proteins of H8 cells and Caski cells were extracted by ProteoExtract? Subcellular Proteome Extraction Kit.②The nucleus proteins were seperated by two-dimensional gel electrophoresis (2-DE). Different expressions of proteins were firstly analyzed by ImageMaster 2D,and then identified by matrix-assisted laser desorption / ionization time-of-flight mass spectrometry (MALDI-TOF-MS ). Finally, the peptide mass spectra identification was performed based on the Swiss-prot databases though Mascot and MS-Fit program.③Immunocytochemistry was used to analyze the protein expressions ofα-tubulin andβ-tubulin in the climbing. Meanwhile western blotting was used to determine the protein expressions ofα-tubulin,β-tubulin andγ-tubulin by semi-quantitative.Results:①We obtained well-resolved,reproducible 2-DE profiles and 30 notable differential nucleus protein spots were detected in 2-DE gels.Among the proteins identified, 6 proteins(INT6,TRA16,PP4C,CDK6,ANR16,ZN211)were up-regulated in Caski cells and 3 proteins(MYST1,LMNA,TIAF1) were down-regulated in Caski cells.②The protein expressions ofα–tubulin,β–tubulin andγ–tubulin were higher in Caski than H8 (all P<0.05).Conclusions:①Nucleus proteins with different expressions are valuable for the study of cervical carcinogenesis mechanisms,and for diagnostic markers and therapeutic targets in cervical cancer .②Centrosome abnormality may be one of the malignant phenotype of cervical cancer cells. Research of microtubulin protein for a target in tumor treatment has a prospective future.
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