Effects Of Curcumin, Demethoxycurcumin And Bisdemethoxycurcumin On Expression Of MMP-2, MMP-9Proteins In Human Hepatoma Cell Line HepG2

Objective: HCC(Hepatocellular Carcinoma, HCC) is a commonmalignancy in the world, accounting for over90%of primary liver cancer.According to2002statistics, the incidence of liver cancer worldwide eachyear about626,000case,598,000cases of death, its incidence ranked NO.6inthe common tumors,while mortality is ranked NO.3.The hepatocellularcarcinoma incidence and mortality of our country is the highest. The numberof new liver cancer each year accounts for more than half of the world, whilethe number of died from liver cancer accounts for about40%-45%of theglobal deaths. In our country, the mortality of liver cancer ranked NO.2.The early clinical symptoms of liver cancer is not clear, insidious onset,but rapid progress. The majority of patients has locally advanced or distantmetastasis when diagnosed. The invasion and distant metastasis of HCCseriously impact the prognosis of liver cancer. Therefore, an effectiveanticancer drugs urgently needed to control the invasion and metastasis ofHCC.The evolution of tumor form cancer in situ to local invasion andmetastasis, the most important condition is the extracellular matrix(Extracellular Matrix, ECM) degradation of the cell. While the degradation ofECM needs a variety of proteolytic enzymes. The matrixmetalloproteinase(Matrix Metalloproteinases, MMPs)is a major extracellularmatrix degrading enzymes. The basic skeleton of the extracellular matrix iscomposed of collagen IV. So far, the gelatinase (MMP-2å'ŒMMP-9) is theonly proteolytic enzyme to degrade collagen IV. The gelatinase can destroythe extracellular matrix and promote cancer breakthrough extracellularmatrix.therefore, in the process of tumor invasion and metastasis, the gelatinase plays an important role.Curcumin-drugs(curcumin,demethoxycurcumin,besidemethoxycurcumin)is a compound extracted from the rhizome of the plant Curcumin.Demethoxycurcumin and besidemethoxycurcumin is the ramification ofcurcumin. All three have similar pharmacological effects, especially inanti-tumor. Research has shown that curcumin and its derivatives can inhibitthe expression of MMP-2and MMP-9with a dose-dependent manner in thehuman HT1080fibrosarcoma cells. Wether the curcuminoids has the sameinhibitory effect on the expression of MMP-2and MMP-9in HepG-2is rarelyresearched. In this study, we will discuss the effect of curcuminoids on theexpression of MMP-2and MMP-9in human hepatoma cells.Method: Cultivate Hepatocellular carcinoma cell line HepG2in general.Distribution of cell cycle was examined through FCM: HepG2administered(24h) with various concentrations of curcumin,demethoxycurcumin andbesidemethoxycurcumin (20,40μmol/L) were measured with flow cytometryand analyzed by software; Expression of MMP-2and MMP-9in HepG2cellsunder the different concentrations of curcumin,demethoxycurcumin andbesidemethoxycurcumin (20,40μmol/L) and differences among the expressionof them treated by curcumin, demethoxycurcumin and bisdemethoxycurcumin(40μmol/L) was detected by Western-Blot.Result:1Analysis by FCM showed that, the HepG2cells treated with differentconcentrations of curcumin,demethoxycurcumin and besidemethoxycurcumin(20,40μmol/L), compared with the control group, with the increasing of theconcentrations, the numbers of HepG2cells increased obviously in G0/G1phase(P<0.05) while the proportion of HepG2cells in S phase and G2/Mphase decreased(P<0.05). Thus the cells were arrested in G0/G1phase; theHepG2cells treated with same concentrations ofcurcumin,demethoxycurcumin and besidemethoxycurcumin (40μmol/L) for24h,the ability of three drugs for arresting cells in G0/G1phase has nodifference. 2After HepG2cells treated with different concentrations of curcumin,demethoxycurcumin and besidemethoxycurcumin (20,40μmol/L) for24h wereexamined by Western-Blot analysis, compared with the control group, theexpression of MMP-2and MMP-9is down regulated after treated withcurcumin, demethoxycurcumin and bisdemethoxycurcumin in the HepG2cells,with a dose-dependent manner(P<0.05).3After HepG2cells treated with same concentrations of curcumin,demethoxycurcumin and besidemethoxycurcumin (40μmol/L) for24h wereexamined by Western-Blot analysis, the efficacy of three drugs to inhibit theexpression of MMP-2and MMP-9is that, the bisdemethoxycurcumin is thestrongest and the curcumin is the weakest.Conclusion:1Curcumin, demethoxycurcumin and besidemethoxycurcumin can inducearrest of the human hepatic carcinoma cell line (HepG2) in G0/G1phase; theability of three drugs for arresting cells in G0/G1phase has no difference.2Curcumin, demethoxycurcumin and besidemethoxycurcumin all candown regulate the expression of MMP-2and MMP-9, thebesidemethoxycurcumin has the strongest efficacy and the curcumin is theweakest.