Proteomic Analysis Of Cellular Response To Anti-peptide From Meretrix Meretrix Linnaeus In Hepatocarcinoma SMMC-7721 Cells

Meretrix meretrix Linnaeus is smooth-shelled marine mussels.It not only is delicious but has medicinal properties. Some researchers have reported that Meretrix meretrix Linnaeus can lower blood sugar and lipid. It is also resistant to oxidation and has immunocompetence. Now more and more researchers are intresting in study of anticancer active of extract from Meretrix meretrix Linnaeus.We purified the extract of Meretrix meretrix Linnaeus and obtained an anticacer component. The methods include grinding the meat of Meretrix meretrix Linnaeus, extracting by using organic solvents and purification by Sephadex G-25. The component obtained can inhibit cell prolifiration markedly. The inhibitory rate on the cell growth was determined to be 50.6%, after cells were treated by 5.0μg/mL of the component. The morphology of hepatocarcinoma SMMC-7721 cells was observed by light microscope. It can be observed that morphology of cells have changed greatly.Studying the effect of the anticancer peptide of Meretrix meretrix Linnaeus on hepatocarcinoma SMMC-7721 cells by using proteomics method is help to study anticancer mechanism.Two-dimensional gel (2-DE) electrophoresis has recently become the preferred method for proteomics and advanced the utility of proteomics in drug discovery process. In this article we established 2-DE technique successfully by optimizing a series of factors, such as sample preparation, electrophoresis parameters, gel′s concentrations and so on.2-DE patterns were obtained and these patterns were analyzied by using 2-DE software-melanie 4. Protein digest by trypsin in gel and MALDI-TOF analysis were carried out in the proper order. Then identify differential expressed proteins by searching protein database.About 136 of the differential expressed proteins were found on two patterns. One is the control cells proteins pattern, the other is cells treated with anticancer peptide of Meretrix meretrix Linnaeus proteins pattern. There are 96 down-regulated proteins and 40 up-regulated proteins after cell treated with anticancer peptide. We choose some proteins which changed markedly and identified them by MS. Finaly 10 proteins were identified successfully. 6 of proteins are down-regulated proteins, and 3 of proteins are up-regulated proteins and 1 protein whose pI was changed. These proteins include cyskeletal protein, zinc finger protein, pressure elicit protein and so on. Discuss fuctions of these proteins and make hypotheses of the machanism of inhibiting cell prolifiration.This research will be useful to further study of the effects of anticancer peptide from Mercenaria on the SMMC-7721 cells.