| Nowadays,urea and some other body fluid analyses have been used to diagnose diseases.Saliva is a kind of exocrine fluid in human bodies,mainly secreted by major salivary glands(parotids,submandibular glands and sublingual glands).There are more than a thousand of proteins,a great amount of mineral salts and water in saliva.Saliva is easy to gain by rapid,simple and non-invasive ways.However,using saliva to make diseases diagnosis has not been wildly accepted.It has been proved that many salivary proteins expression changes are related to systemic and oral diseases,such as diabetes mellitus,Sjogren syndrome;periodontitis and oral squamous cell carcinoma.Parotid,as one of the three major salivary glands,can be identified with many kinds of tumors.So,will there be any detectable abnormalities in salivary proteins when parotid is invaded by tumors?In order to investigate the salivary protein expression abnormalities in parotid pleomorphic adenoma(PMA)and parotid malignant tumors(PMT),we performed a comparative salivary proteomic analysis by Liquid chromatography electrospray ionization tandem mass spectrometry(LC-ESI-MS/MS)among PMA,PMT and normal groups.PURPOSES:1.Find some peotein biomarkers by analyzing the parotid saliva proteome of normal and tumor-invaded individuals to reflect the present of parotid thmors.2.Demonstrate that the importent proteins indentified by LC-ESI-MS/MS realy exist in saliva.3.Find some representative proteins by comparing the salivary proteins of different parotid tumor individuals,to help to certify the properties of tumors.METHOD:A total of 15 individuals were involved in LC-ESI-MS/MS to define the candidate proteins,among which 5 were PMA,5 were PMT and the other 5 were normal individuals.For control group collect a random side,while the tumor group collect the affected side.Extract,purify,ultrafiltrate and concentrate,lyse,desalt the salivary proteins,and then under go the LC-ESI-MS/MS analysis.Analyse the obtained data and find the differential proteins.What's more,we detected the identified galectin-7 protein,which is significantly up-regulated in PMA group,by ELISA assay kit in more samples.RESULTS:The results show that 21 proteins are significantly changed(up-regulated or down-regulated more than 1.5-fold)in tumor groups,including galectin-7,SPARCL1,Azurocidin,ARPC4-TTLL3,Lipocalin 1,and cystatin C.Above which galectin-7 is up-regulated in PMA group compared with control and PMT groups,SPARCL1(Hevin)is down-regulated in PMT group compared with control and PMA groups,Azurocidin and ARPC4-TTLL3 are up-regulated in PMA and PMT groups compared with control group,while Cystatin C and Lipocalin 1 are down-regulated in PMA and PMT groups compared with control group.A further ELISA detection was performed to identify the galectin-7 expression abnormality and the independent-samples nonparametric test shows that galectin-7 is significantly up-regulated in 18 PMA individuals compared with 6 normal ones(P=0.002)and 5 PMT ones(P=0.008),while not significantly different(P=0.193)between control and PMT groups.CONCLUSION:1.LC-ESI-MS/MS analysis can select the significantly changed proteins in saliva of parotid pleomorphic adenoma patients.2.The salivary galetin-7 expression abnormality in PMA patients,compared with control and PMT patients,can be certified both by ELISA assay kit and LC-ESI-MS/MS analysis,while there is no difference of galectin-7 expression between control and PMT groups.3.Our results indicate that galectin-7 may serves as a potential factor in preoperative non-invasive identification of parotid tumors by saliva.4.Others abnormally expressed proteins need to be certified by futher studies. |
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