Anti-tumor Activity Of Curcumin Derivative C1207 And Its Mechanism

Objective To study the anti-cancer activity of curcumin derivative C1207 and its mechanism.Methods(1) The MTT assay was used to detect the role of proliferation inhibition of C1207 on BEL-7402, Hep G2, HCT-116, Hela,K562, K562/G01,HL-60 and BT474 tumor cell lines;(2) The method of colony formation: to observe how the C1207 influenced the colony formation of liver cancer cell BEL-7402 and Hep G2;(3) PI single staining inspected the effect of C1207 on the cycle process of hepatic carcinoma cells BEL-7402 and Hep G2 via the FCM;(4) Used Annexin V-FITC/PI staining to detect the apoptosis effect of C1207 on hepatoma cell-lines BEL-7402 and Hep G2 through the FCM(Flow Cytometry);(5) JC-1 staining detected the influence of C1207 on mitochondrial membrane of hepatic carcinoma by the FCM;(6)Western Blotting analysis was applied to detect the effect of C1207 on expression of the apoptosis-related and autophagy-association protein;(7) MDC staining tested the influence of C1207 on the autophagy of the hepatic carcinoma cell by the HCA(High content Analysis);(8) Fluorescence quenching detected the interaction of C1207 with different fragments of Hsp90 protein.Results(1) After the treatment for tumor cells with the C1207 s for 24 h and 48 h,C1207 could signally inhibit the proliferation of the multiple tumorcells, such as BEL-7401, Hep G2, HCT-116, Hela, K562, K562/G01, HL-60 and BT474 in dose-dependent depression. The IC50 value of the most tumor cells was less than10μM.(2) The C1207 s cloud inhibit significantly the colony formation of the BEL-7402 and Hep G2 cell.(3) The cell cycle experiment showed BEL-7402 cell mainly blocked in G2/M phase, while the Hep G2 cell blocked S phase.(4) Use the C1207 with the concentration of 0, 4, 8,12μM to treat BEL-7402 and Hep G2 cells respectively 24 hours.With the increase of drug concentration, the proportion of apoptosis was on the rise.(5) The C1207 s with the concentration of 0, 4, 8, 12μM were respectively applied to BEL-7402 and Hep G2 cell 12 hours later, with the increase of drug concentration, the JC-1Red fluorescence intensity decreased, while the JC-1 Green fluorescence intensity gradually enhance and the trend of mitochondrial membrane potential lowered.(6) C1207 could influence the expression of apoptosis-associated proteins of hepatic carcinoma cell.(7) C1207 could induce the autophagy of hepatic carcinoma cell. As the increasing drug concentration and action time,the autophagy became more intensified step by step.Hepatic carcinoma cell treated the C1207 with the concentration of 8μM 24 hours later,the autophagy reached the summit and began to lower gradually 48 hours later.(8) The C1207 s with different concentration were applied to hepatic carcinoma cells BEL-7402 and Hep G2 24h: the ratio of LC3-II/I augmented little by little, namely, the activity of autophagy strengthened gradually, showing the feature of dose-dependent.(9) C1207 could interact with N-Hsp90,M-Hsp90,C-Hsp90 protein, and the combination between C1207 with C-Hsp90 was the strongest.Conclusions(1) C1207 could inhibit the proliferation of tumor cell lines in dose-dependent.(2) C1207 could stagnate the hepatic carcinoma cell in different phases.(3) C1207 could induce apoptosis of hepatic carcinoma and lower the mitochondrial membrane potential.(4) C1207 could affect the expression of apoptosis-related proteins.(5) C1207 could induce the autophagy of hepatic carcinoma cell.(6) C1207 could interact with N-Hsp90,M-Hsp90,C-Hsp90 protein.