| Aims: Allicin is a key ingredient isolated from garlic extract, previous studies haveshown that allicin can significantly inhibit ovarian cancer H08910and OVCA-3growth,and induction of apoptosis. However, allicin induced ovarian cancer cell lines, themechanism is not very clear. In this study, human ovarian carcinoma SKOV3cells asexperimental subjects, and to clarify the allicin-induced apoptosis of ovarian cancermechanism.Main methods: The proliferation inhibition rates of SKOV3cells after varioustreatments were examined by CCK-8’assay; After the treatment of SKOV3cells by allicin,apoptosis rate was determined with Annexin V-FITC/PI double staining by flow eytometry;Flowe-ytometry was used to observe the effect of allicin on cell cycle distribution ofSKOV3cells; and activation of the signaling pathway was screened by human phospho-kinase array; the activated and its related proteins were further confirmed by immunoblots.Key findings:(1) Different concentrations of allicin for different time inhibit theproliferation of SKOV3cells: CCK-8assay results showed that SKOV3cells’growth wassignificantly inhibited by allicin in a time–and-dose dependent. Differenr concentrationsof allicin in SKOV3cells for24,48,72h, the inhibition rate differences were significant(P<0.05). In the efficiency equation derived, the IC50were22.23μg/ml in48h.(2) Allicinon SKOV3cell cycle:12.5,25μg/ml concentration of allicin in SKOV3cells after48h,cell cycle changes. Each experimental group to reduce the percentage of cells in G1phase,no significant changes in the percentage of S phase cells, increased percentage of cells inG2phase, cells arrest at the G2phase role significantly, compared with control group wasstatistically significant (P <0.01).(3) Allicin on apoptosis of SKOV3cells:12.5,25μg/mlconcentration of allicin for48h, cells apoptosis rate with increasing drug concentrationincreased, the early and late apoptosis rate (%), respectively,7.37±1.21,15.87±1.65,with the control group (2.33±0.42) were significantly increase (P <0.01), suggesting thatwithin the experimental range, the allicin-induced apoptosis and drug concentration waspositively correlated; After treatment with the20μM JNK inhibitor (of SP600125) in15min, compared with25μg/ml allicin group, apoptosis rate was significantly reduced (P<0.01).(4) The protein chip: After treatment with12.5μg/ml allicin for24hours, thephosphorylated protein chip display, P53(S46), AKT (T308) and c-Jun (S63) and otherapoptotic pathway is activated.(5) Allicin on the expression of JNK pathway: Aftertreatment with (0-100μg/ml) allicin for15min, the phosphorylation of JNK protein in increased significantly with increasing concentrations; After treatment with25μg/mlallicin at different times, the strongest of JNK phosphorylation in15min; treat withSP600125, JNK phosphorylation was significantly inhibited. That allicin-inducedapoptosis in SKOV3cells related to the JNK pathway.(6) Allicin on the mitochondrialpathway: After treatmen with25μg/ml allicin in SKOV3cells for (0,6h,12h), in thecytoplasmic protein, Bax protein expression is diminished, Cty C protein expressionenhanced; just the opposite in mitochondrial protein. Join SP600125in the cytoplasmicprotein, Bax protein expression increase compared with12h of allicin group, theexpression of Cty C protein decreased; while the opposite in the mitochondrial protein.That JNK activation, including activation of Bcl-2family, triggering the activation ofmitochondria-mediated signaling pathway, lead to large accumulation of Bax translocationand Cyt C release.Conclusion: Our data show that allicin can inhibit cell proliferation and induce cellapoptosis, and arrest the cells in G2phase, this effect may be related to the JNK pathwayactivation and Bax translocation, causing the release of cytochrome C related. |
PDF Full Text Request