Effects Of 5-ally-7-gen-difluormethylenechrysin On The Growth And Apoptosis Of Human Lung Cancer A549 Cell Line

Objective:To investigate the effects of 5-allyl-7-gen-difluoromethyllene-chrysin (ADFMChR) on selective inhibition of the growth and induction of apoptosis in human lung cancer A549 cell line.Methods:Human lung cancer A549 cell line and Chinese hamster lung epithelial CHL cell line were cultured in vitro.Colony formation assays on plate and in soft agar were used to detect the anchorage-dependent and anchorage-independent growth of cells.Flow cytometry(FCM) using PI staining was used to analyse the apoptosis rate of cells.DNA agarose gel electrophoresis was used to confirm induction of cell apotosis.Results:The results of colony formation assay on plate showed that the growth inhibitory rate of A549 cells after treatment with ADFMChR at various concentrations of 0.1,1.0,10.0,100.0μg/mL for 48h was 16.3%, 38.9%,65.6%,96.7%respectively.ADFMChR significantly suppressed the anchorage-dependent growth of A549 cells in a concentration dependent manner,and its IC₅₀ was 1.6μg/mL.The efficacy of inhibition with ADFMChR was stronger than that of chrysin(IC₅₀ was 9.3μg/mL) and was similar to Tax(IC₅₀ was 1.4μg/mL).The growth inhibitory rate of CHL cells was 14.7%,29.6%,52.4%,74.1%respectively after treatment with ADFMChR at various concentrations of 0.1,1.0,10.0, 100.0μg/mL for 48h.ADFMChR significantly suppressed growth of CHL cells in a concentration dependent manner,and its IC₅₀ was 7.8μg/mL. The cytotoxicity of ADFMChR to CHL cells was lower than that of Tax (IC₅₀ was 3.1μg/mL) and was similar to lead compound chrysin(IC₅₀ was 8.5μg/mL).The data of colony formation assay in soft agar indicated that the growth inhibitory rate of A549 cells after treatment with ADFMChR at various concentrations of 0.1,1.0,10.0,100.0μg/mLfor 48h was 23.6%, 42.8%,66.1%,86.7%respectively.ADFMChR significantly suppressed the anchorage-independent growth of A549 cells in a concentration dependent manner,and its IC50 was 1.7μg/mL.The efficacy of inhibition with ADFMChR was stronger than that of lead compound chrysin(IC₅₀ was 10.6μg/mL) and was similarto Tax(IC₅₀ was 1.5μg/mL).Flow cytometry(FCM) using PI staining found that the apoptosis rate of A549 cells after treatment with ADFMChR at various concentration of 1.0, 10.0,100.0μg/mL of for 48h was 12.0%,19.3%,28.0%respectively,and the apoptosis rate of A549 cells after treatment with ADFMChR at 10.0μg/ml for various time 24h,48h,72h was 4.5%,19.3%,18.8% respectively.ADFMChR significantly induced apoptosis of A549 cells,in a concentration and time dependent manner.The apoptosis rate induced by ADFMChR was higher than that of chrysin and similar to Tax.The apoptosis rate of CHL cells was 10.2%,14.3%,26.9%respectively after treatment with ADFMChR at various concentrations of 1.0,10.0, 100.0μg/mL for 48h,and the apoptosis rate of CHL cells was 4.8%,14.3%,14.6%respectively after treatment with ADFMChR at 10.0μg/mL for 24h,48h,72h.ADFMChR significantly induce apoptosis of CHL cells in a concentration and time dependent manner.The apoptosis rate of ADFMChR to CHL cells was lower than that of Tax and was similar to chrysin.DNA agarose gel electrophoresis indicated that ADFMChR significantly induced apoptosis of A549 cells,the result also show that ADFMChR significantly induced apoptosis of CHL cells.Conclusion:1.ADFMChR significantly inhibits the anchorage-depent growth of human lung cancer A549 cell line in vitro.2.ADFMChR significantly inhibits the anchorage-indepent growth of human lung cancer A549 cell line in vitro.3.ADFMChR effectively induced apoptosis of A549 cells.4.Comparison with the inhibitory efficacy of A549 cells,the cytotoxicity of ADFMChR to CHL cells was lower.