Proetomics And Inhibition Effect Of Curcumin In Vitro About Ovarian Cancer

1.Analysis of Proteins with Differential Expression Induced by DDP in Human Ovarian Adenocarcinoma Cell Line COCProteomics is focus point in postgenome age．It provide new thinking method of the research of severe diseases of human beings．Human ovarian cancer is one of the most maligmant tumors in gynecopathy with a high mortalityrate,diagnosis and therapeutic effect isn't excellent.In this present,human ovarian cancer cell line COC1 and its cell sublineinduced by Diamminedichloroplatinum(DDP)were used as the compared sy- stem.We investigated for biomarkers of ovarian cancer with proteomics ways At first,we studied the effect of the drug dose on the cellular cycle and apoptosis rate by using flow cytometry．The cytotoxicity of DDP correlates with cellular arrest in the G2/M phase.50?g·ml of DDP can result in the change of cell apopt osis and cell cycle obviously(P<0.01)．Then,COC1 cell and COC1 induced by 50?g·mL of DDP cell line were separated and analyzed by high resolution two dimensional eletrophoresis(2-DE) techniques.Through considering the restriction of experimental pI vaule and molecu ar weight(MWt) in 2-DE maps and searching for the peptide in the bio-databade according to PMF,the peptide mass fingerprint(PMF) of the enzymatically-digested proteins that express diffenerce obviously in 2-DE images were identified by using matrix assisted laser desorption/ionization mass spectro I-TOF-MS) technique. Finally 11 protein spots in the gels were identified.Comparing the ovarian cancer cell COC1 and the treated by DDP,7proteins among of them(all for ovarian cancer cell COC1 in response to DDP) have a significant different Mean While, expression levelsprotein are different 4 proteins．Among these proteins identified,5proteins are"molecular chaperones"proteins,including of HSP70,HSP90,BiP,acute T cell leukemia associated with protein-1(TAL-1),Prohibitin．These molecular chaperones proteins maybe accociated with tumorigenesis, growth, apoptosis. 4 proteins associated withenergy metabolism have not be sure that they relate to tumor,including of ATP synthase,Iodotyrosine deiodinase,Human Dutp Pyrophosphatase, Complex with Dudp and Inositol hexaphosphate kinase2．The function of another 2 proteins have been unkown．Molecular chaperones proteins can protect cell and resist chemotherapeutics in the phase of apoptosis．Prohibitin (phb) gene is one of tumor-suppressing genes play roles of anti-proliferation and anti-tumor．Prohibitin (PHB) coded by it reside in not only mitochondrial inner membrane,but also intranuclear．It has the function of negativity trans- criptional control．PHB produce a marked effect in the phase of G1-S specifily In this study,PHB only has a little expression in COC1,while a high expression in COC1subline induced by DDP．It confirm thoroughly the common result with the study of predecessor that PHB has the function of resisting tumor.TAL-1 is one alkalinity transcription factor of coiling ring coiling family,it has been indicated that has related with hematopoietic cell proliferation,cell differentiation and leukemia．It is one of oncoprotein．In this study,TAL-1 is significant up-regulated expression in COC1 subline induced by DDP. It needed to research deeply therelationship between TAL-1 and proliferation and apoptosis of ovarian cancercell． In the word,we discover some proteins that correlate with cell cycle and apoptosis of ovarian cancer cell．The result provide new theory evidence for realing the pathogenesy of ovarian tumor and exploring new antitumor drug．2. Inhibition Effect of Curcumin on Ovarian Cancer Cell Line COC1 and COC1/DDP in VitroCurcuma is the rhizome of Curcuma longa Linn,Curcumin(Cur) is the main component of curcuma,belong to natural phenols anti-oxidant,frequently used conimex and edible pigment. Cur has the features of safety and no toxic side effect. As antitumor agent,Cur has become one of popular projects in the field of tumor prophylaxis and thrapy．In this study,we apply MTT chromatometry,high performance liquid chromatography(HPLC) to probe the synergistic inhibition of Cur combining with diamminedichloroplatinum(DDP)on COC1,reverse multiple drug resistance effect of Cur on COC1/DDP and sensitization of Cur on COC1 and COC1/DDPto DDP are studied by using MTT and HPLC．This results show that the synergy of Cur and DDP effect on COC1．5?g·mL-1 of Cur combining with DDP 2.5?g.ml-1-10?g.mL-1 produced simple addition or potentiation effect；Cur10μg·mL-1 combining with DDP 2.5μg·mL-1～10μg·mL-1produced simpleaddition or antagonistic effect．Meanwhile Cur can enhance the sensitivity of ovarian cancer tend cell line COC1 to DDP,the multiple is 1.5．when Cur5?g·mL-1,completely reverse the drug resistance on ovarian cancer multiresistant cell COC1/DDP,multiple is 18．It has the same effect with verapamil(VRM) as 20?g·mL (P>0.05)．Meanwhile,Cur increase the sensitivity of ovarian cancer multiresistant cell line COC1/DDP,multiple is equal with reversed multiple．Another,the time of drug initiation of DDP can be shorted by Cur from 18h to12h．There are no celluar accumulation in COC1 and COC1/DDP because of Cur combining with DDP(P>0.05)．In summary,Cur has the powerful antitumor activity．It has the coordinate repression of Cur combining with DDP on COC1,only low concentration Cur can reverse the multiresistant on COC1/DDP,meanwhile it can enhance the sen itivity of COC1 and COC1/DDP to DDP,decrease the dosage of DDP,shorten time of drug initiation．Because of no toxic side effect,Cur can lessen thesevere system damage in order to lessen the suffering of patient．On the other hand,there are no celluar accumulation in COC1 and COC1/DDP because of Curcombining with DDP,it noted that multiresistant of COC1/DDP maybe have nothing to do with membrane protein like P-gp．The exact mechanism of multiresistant need research deeply．If the shortcomings of low dissolubility and metabolism rapidly in vivo could be overcome by new technique and method,Cur will become a new safety and high-performance reversal agent of drug resistance and antitumor chemicals．This research may be can develop a new safety and high-performance clinical treatment scheme．...