A Comparative Proteomic Study Of Cervical Carcinoma Tissues With Different Concurrent Chemoradiotherapy Sensitivity

Objective: To investigate the proteomic differences between thehigh-sensitivity (HS) group and the low-sensitivity(LS) group of cervicalcarcinoma treated by concurrent chemoradiotherapy and confirm theconcurent chemoradiotherapy sensitivity associated proteins in interme-diate stage and advanced cervical carcinoma.Methods: The fresh carcinoma tissues were collected from 10untreated cervical carcinoma patients and preserved in the -80℃refrigera-tory. The tissues were classified into two groups: high sensitivity group(HS) and low sensitivity group (LS) , according to their response toconcurrent chemoradiotherapy. In the first part of our experiment, proteinseparating was performed by using two-dimensional gel electrophoresis(2-DE) with Amersham 18cm linear pH 3-10 immobilized pH gradient(IPG) strips. The images of the gels were acquired by the scanner and thenanalyzed by using PD-quest7.0 software to find the differentiallyexpression protein-spots in each group. Then the differentially expressedprotein-spots was incised from the gels and digested by trypsin .Thepeptide mass fingerprintings(PMF) was acquired by matrix assisted laserdesorption/ionization time-of-flight mass spectrometry(MALDI-TOF-MS)and the proteins were identified by data searching in the Mascot-database.Four differentially expression proteins were assayed by Western Blot andimmunohistochemical method. The clinico pathological significance of differentially expression proteins were analysized.Results: Most of the gels were clear and successfully analyzed byPD-quest7.0 software. Most of the protein-spots concentrated on the area of20～100KDa (Mw) and pH4～8. The average number of the protein-spotswas 781±74 in high-sensitivity group and 766±52 in the low-sensitivitygroup. The match rate was 87.6 % between two groups. 9 high expressionproteins were found in HS group which were low expression in LS groupincluding Hemoglobin subunit beta, Caspase-14 precursor , Calmodulin-like, S100-A9 protein/MRP-14, galectin-7, HSKERC4 , keratin 19 andActin; 10 high expression protein were found in LS group which werelow expression in HS group including Anti HBs antibody light-chain Fab,Lamin-B1, WARS protein, Flavin reductase , Glutamate dehydrogenase1, nuclear matrix protein 238, Retinal dehydrogenase 1, AF165172,Subunit of replicative DNA polymerase and HSP70. The higherexpression of HSP70 and NMP238 proteins in LS group and S100-A9protein and galectin7 in HS groups were further confirmed by western blotand immunohistochemical assay.Conclusions: The 2-DE gets image of high-sensitivity (HS) groupand the low-sensitivity(LS) group of intermediate stage and advancedcervical carcinoma tissues treated by concurrent chemoradiotherapy weresuccessfully acquired, 19 differentially expressed proteins were identifiedby mass spectrometry; Some differentially expression proteins betweenthe two groups were further confirmed by western blot and immunohis-tochemical assay. These proteins may be associated with sensitivity ofconcurrent chemoradiotherapy and expected to be candidate biomarker forprediction of sensitivity of concurrent chemoradiotherapy.