Experimental Study In The Growth Inhibiting Effect Of Pomiferin On Tumour Cell

Objective:Isoflavones is widely found in nature,a class of compounds,many isoflavones have been proven to a certain degree of experimental anti-tumor activity and prevention of cancer is as safe and low toxicity.A large number of studies have shown that apoptosis is induced by flavonoids,one of the anti-tumor mechanism.This experiment will study cell proliferation in vitro inhibitory aspects of orange from the Mulberry(Osage Orange) in the isolated isoflavones Pomiferin(structure Figure 1) on MCF-7 breast cancer,SKOV-3 ovarian cancer tumor cell proliferation inhibitory effect of and from the best concentration of the drug screening,further studies Pomiferin effect on the tumor cells cell cycle,apoptosis,apoptosis inhibitory factor and pro-apoptotic factors such as the impact of several aspects.Pomiferin expectations in the role of tumor prevention and treatment to provide a new clue and means.Method:MTT assay with different concentrations of Pomiferin(1.5mg/L,3mg/L, 6mg/L,12mg/L,24mg/L) of MCF-7 breast cancer and ovarian cancer SKOV-3 tumor cell proliferation inhibition,inhibition of tumor Pomiferin selected the best concentration of cells;Pomiferin flow cytometry on cell cycle and apoptosis induced by the results;RT-PCR assay After the tumor cells Pomiferin role Survivin,Bax and bcl-2 in three and apoptosis-related gene expression at the mRNA;immunocytochemical detection Pomiferin the role of inhibitor of apoptosis protein Survivin expression changes.Results:Pomiferin of five tumor cell lines have a wide range of inhibition.On MCF-7 breast cancer and SKOV-3 ovarian cancer Pomiferin inhibit tumor growth in the most effective concentration range of 6-12mg/L;cycle showed MCF-7 breast cancer and SKOV-3 ovarian cancer in the 10mg/L Pomiferin the role of 24h,48h after cell cycle arrest in G₂/M phase and apoptotic peak appeared;apoptosis detection showed that MCF-7 breast cancer 10mg/L Pomiferin after early apoptosis rate of 25.14%,apoptosis rate of cells in advanced 24.12%,SKOV-3 ovarian cancer 10mg/L Pomiferin after early apoptosis rate of 38.12%,the cell apoptosis rate of 46.78% advanced;RT-PCR detection of apoptosis inhibitory factor and pro-apoptotic factor mRNA expression was found in the 10mg/L Pomiferin role in the control group cells with little change in Bax,after 10mg/L Pomiferin much lower than the Bcl-2,Survivin mRNA expression of the control group;immunohistochemical detection of tumor cell apoptosis inhibitor protein Survivin in the cytoplasm of both the expression of nuclear,10mg/L Pomiferin after Survivin protein expression was significantly weaker than the control group.Conclusion:Pomiferin can inhibit MCF-7 breast cancer and SKOV-3 ovarian cancer cell proliferation;and inhibition of cell cycle in G₂/M phase,pomiferin role obviously the latter two cell early apoptosis and late apoptosis;pomiferin can significantly reduced Survivin and Bcl-2 mRNA expression,but Bax mRNA expression was no signifycant change;Pomiferin significantly reduced apoptosis inhibitor protein Survivin expression.Pomiferin mechanism may be induced by the reduction in inhibitor of apoptosis, tumor cell cycle arrest in G₂/M phase of tumor cells to achieve growth inhibition.