HPV永生化细胞系与SiHa细胞系蛋白质表达差异的质谱分析

Mass spectrometry analysis of differentially expressed proteins between HPV immortalized cell line and SiHa cell line

目的分析人乳头瘤病毒(HPV)永生化细胞系与Si Ha细胞系之间的蛋白质表达差异,探讨从HPV感染到宫颈癌发生间的变化。方法高效液相色谱(LC)与质谱(MS)联用对HPV永生化细胞系与Si Ha细胞系的蛋白质进行鉴定,然后通过蛋白质组学和生物信息学分析两种细胞系之间的蛋白表达,及一种新型翻译后修饰-琥珀酰化修饰的差异。结果在两种细胞系中,质谱鉴定到的蛋白总数为10 691个,琥珀酰化修饰位点共6 342个;非标定量分析共发现572种差异表达蛋白。HPV永生化细胞系中高表达的273种蛋白多集中在疾病的相关通路,而宫颈癌细胞系中高表达的299种蛋白多集中在代谢通路和核糖体通路。琥珀酰化修饰无差异,但发现一个新的琥珀酰化修饰位点——血清和糖皮质激素激酶1(SGK1)K41琥珀酰化。结论与HPV永生化细胞系相比,Si Ha细胞系代谢能力及翻译蛋白能力更强;而HPV永生化细胞中高表达蛋白则大多与疾病相关,且更易发生恶化。

Objective To explore the mechanism by which human papilloma virus（ HPV） induces cervical cancer and to identify proteins differentially expressed between HPV immortalized cells and Si Ha cells. Methods Differentially expressed proteins between HPV immortalized cells and Si Ha cells were identified by high performance liquid chromatography and mass spectrometry,and analyzed by label-free quantity software and bioinformatics approaches. Results A total of 10 691 proteins were identified and 6 342 succinylation sites were found; label-free quantification analyses indicated that 572 proteins were differentially expressed between the two types of cells; bioinfomatic analysis showed that273 proteins in HPV immortalized cells were mostly associated with disease pathway,while 299 proteins in cervical cancer cells were mostly involved in metabolic and protein-produced pathway. Although succinylation modifications had no differences,a newsuccinylation site at SGK1 K41 was found. Conclusion Compared with HPV immortalized cells,Si Ha cells demonstrate higher level of metabolism and stronger protein-synthesis,while most up-regulated proteins in HPV immortalized cell are associated with particular diseases.