Quantitative Peptidomics Analysis Of Colchicine And Cisplatin-induced Apoptosis In Hela Cells

Peptidomics is a technology of qualitative and quantitative analysis of peptides in biological samples.The applications of peptidomics include identifying peptide biomarkers of disease,identifying bioactive peptide and investigating proteolytic regulation of bioactive peptides.In this study,we applied peptidomics to examine the changes of peptides within the Hela cells after treatment by colchicine and cisplatin which induce cell apoptosis.Cell apoptosis is an important physiological processes in life,which play an important role in maintaining homeostasis,immune function and growth and development.Therefore,it is of great significance to study apoptosis process.Colchicine is an alkaloid that has been used to treat gout and cancer for a long time,and it can induce cell apoptosis by inhibiting microtubule polymerization which has been well documented.Cisplatin is a heavy metal complex,and it can inhibit the replication of DNA by binding to DNA and induce apoptosis.While there have been many ways to explore the apoptosis mechanism induced by these two kinds of medicine,the impact of colchicine and cisplatin on intracellular peptides has not been previously reported.Moreover,attempts to use the information on these peptides from cells for apoptosis mechanism studies have been fairly limited to date,largely due to the lack of awareness that intercellular peptides may contribute to the biological effects of many physiological process including apoptosis.In this study,quantitative peptidomics was used to examine peptides changes of treating Hela cells with 200 ?M colchicine and 50 ?M cisplatin for 24 h.The results are as follows:1.The cell viability was determined with CCK 8 assay.Cell viability was decreased with the increase of drug concentration.2.Hela cells were treated by 200 ?M colchicine for 24 h.A total of 203 peptides were identified,and these peptides represent naturally-occuring fragments of 101 different proteins.Among the peptides,84 peptides were differentially expressed.Among the differentially expressed peptides,3 of these peptides represent N-terminal protein fragments and 24 represent C-terminal protein fragments.Other 57 peptides are the internal fragments of the precursor protein.3.Hela cells were treated by 50 ?M cisplatin for 24 h.A total of 61 peptides wereidentified,of which 34 peptides displayed significant changes.These peptides represent naturally-occuring fragments of 26 different proteins.Among the differentially expressed peptides,only one of these peptides represented N-terminal protein fragments and 14 represented C-terminal protein fragments.Other 19 peptides were the internal fragments of the precursor proteins.4.For the same peptides identified by the treatment of two drugs respectively,some peptides were down-regulated in both experiment and some were only significantly changed in one experiment.There weren't same peptides up-regulated.The different mechanism of the two drugs may contribute to the results.In this study,we have applied an effective analytical platform for comprehensive analyses of endogenous peptidomes during apoptosis,which is a new approach for apoptosis studies,and also indicate that colchicine and cisplatin alter the balance of intercellular peptides,which may contribute to the biological effects of these drugs.