Anticancer Activity And Mechanism Of Actions Of Glycyrrhetinic Acid Compounds And Licorice Chalcone Derivatives In Human Heaptocarcinoma Bel-7402Cells

Objective: To screen active monomers that have higher anticancer activity aboutGlycyrrhetinic acid compounds and Licorice chalcone derivatives; investigate theirinhibitory activity towards human hepatoma Bel-7402cancer cells proliferation and thegene (p53、Bax、bcl-2) expression regulation at the viability, life-cycle and apoptosisinducing effect; investigate also the cytotoxicity on normal hepatocyte(Chang liver) invitro. Methods: This research used normal hepatocyte (Chang liver) and humanhepatocarcinoma cell (Bel-7402) as the in vitro experimental models. The cytotoxicity andanticancer activity of the prepared samples or compounds were screened by MTT assay.The state of pro-apoptotic cell cycle and the apoptosis-promoting effect of the compoundson hepatocarcinoma (Bel-7402) cells were investigated, within flow cytometry technology.Finaly, the regulation hability to gene (p53、Bax、bcl-2) expressions of these compoundswere investigated by the method of Western Blot. Results:1. We found that the flavonoidsextracts A and B and monomer Glabridin from Glycyrrhiza glabra has no significantinhibitory effect on Chang liver cell. Following the increasing concentrations at the rangeof5-500μg/ml, the flavonoids extracts of licorice plant showed the inhibitory rate towardscancer Bel-7402cells was increased gradually, among them, the inhibitory effect ofmonomer glabridin is strongest.2. The glycyrrhetinic acid cerivatives18α-GA、18β-GAand18βGA-5FU showed relatively weakly cytotoxcic effect towards normal chang livercell; but significant inhibitory effects to the proliferation of human hepatoma Bel-7402cells. Furthermore, at the concentration range of80-160μmol/l, the cancer cells apoptosiseffect of GA-5FU and5F reached57-59.1%and16.7%-20.1%.3. The inhibitory effects ofthree licochalcone derivatives (ILG,3-MO-ILG,3OH-ILG) to Bel-7402cancer cells andnormal hepatic Chang liver cells were followed the increased concentrations at the rangeof5-320μmol/l, among them, the inhibitory rate of ILG and3-MO-ILG to Bel-7402 cancer cells was most obvious. The two chalcone derivatives was demonstrated stronglyapoptosis effects towards human hepatomacancer Bell-7402cells. At the concentrationrange of40μmol/l-160μmol/l, their apoptotis rates were reached10.6%-79%and10.1%-93.7%detected by flow cytometry technology, respectively. In the apoptosismechanism study, we found that ILG and3-MO-ILG could encourage the apoptosis byretardation to G2/M phase, at the concentration of IC50. The western Blot studyindicated that the more active chancole derivative3-MO-ILG promote Bel-7402apoptosisby possible mechanism by uper regulation of p53and Bax, and by down regulation ofBCL-2expression. Conclusion:1. Licorice flavonoids extract A and B, and the activemonomer glabridin have weakly citotoxcicity to normal chang liver cells; and theyshowed significant inhibitory effects to hepatoma Bel-7402cells proliferation.2. Amongthe glycyrrhetinic acid compounds, the18α-GA,18β-GA and GA-5FU were shownsignificant inhibitory effect on cancer Bel-7402cells.3. Licorice chalcone derivative3-MO-ILG showed strong inhibitory activity to cancer Bel-7402cells within promotingcell apoptosis by retardation to G2/M phase. The possible mechanism of this action is that,chalcone compound3-MO-ILG could perform the anticancer activity by up-regulation ofp53and Bax expresstion, and by down-regulation of bcl-2-related mitochondrial pathwayto promote apoptosis of hepatoma Bel-7402cells.