Comparative Proteome Analysis Of HPV-infected Cervical Specimens And The Difference Between The High- And Low-risk Genotypes Of HPV

Objective To analysis comparative Proteome of HPV-infected Cervical Specimensand investigate different expression Between High and Low Genotypes. Thedifferential expression protein spots were identified by Mass Spectrometry.Methods HPV DNA was genotyped based on the amplification of HPV DNA bypolymerase chain reaction with a pair of universal primers, MY09/11, in 42 clinicalspecimens, and subsequently subject to either sequencing or a liquid chiphybridization method, luminex. The cervical specimens were divided into twogroups (CIN and CA) according to the genotypes and were analyzed the differentionof the two-dimensional gel electrophoresis (2-DE) in this study with comparingnormal tissues. The differential expression protein spots were identified by MassSpectrometry.Results1. HPV DNA was genotyped based on both sequencing and a liquid chiphybridization method, luminex in HPV infective specimens of 31 cases of CA and11 cases of CIN. (1) HPV in the specimens of CA and CINⅠwas 6, 11 etc.low-risk genotypes, Sequencing was an single peak. HPV in the specimens ofCINⅡ～Ⅲwas 16, 18, 33, 58 etc. high-risk genotypes or mixed type, Sequencingwas an single peak or miscellaneous peak. (2) HPV Infective Genotypes was thesame in one patient.2. Three specimens were composed mixed pooling in each group. 2-DE for the CA,CIN and normal tissues in each group was repeated three times respectively andwas compared differential protein between each of two groups. The expressionof 63 proteins were found significantly between CA specimens and normal tissues, 51 spots were upregulated, 12 spots were downregulated in CAspecimens.The biggest difference between the two groups is 20.2939 times. Theexpression of 47 proteins were found significantly between CIN specimens andnormal tissues, 21 spots were upregulated, 26 spots were downregulated in CINspecimens. The biggest difference between the two groups is 11.2506 times. Theexpression of 39 proteins were found significantly between CIN specimens andCA specimens,15 spots were upregulated,24 spots were downregulated in CINspecimens. The biggest difference between the two groups is 4.24857 times.3.26 differential spots were selected and analyzed, 22 peptide mass fingerprints(PMF) maps were obtained by MALDI-TOF-MS. Recognized the apexes withMascot distiller software, and search for NCBIrtr database, as a result, 18 proteinswere preliminarily Identified. There were correlated cell metabolism, signaltransduction, cell secreting, cell cytoskeleton contruction, cell proliferation andapoptosis.Conclusions The expression maps of proteome in warts, CIN specimens ofcervical tissues and normal cervical tissues are preliminarily established by 2-DEtechnique, and the differential proteins between each of two groups is obtained.Some of differential proteins are identified through MS and bioinformatics technique.Some of them are probably closely related with occurrence and development ofcervical cancer. The results provide useful data for setting up database of HPVinfection in cervical tissues proteome. Further research is needed on whether theseproteins could be used as diagnostic marks or therapeutic targets.