The Effect Of Phyllanthus Extracts On Hepatoma Cells

Objective:To investigate Phyllanthus extracts at different concentrations doses acts on cultured human hepatoma cell line BEL-7404, groping their liver cancer cell killing effect induced apoptosis and the optimal dose, and further research from the molecular level Phyllanthus inhibition of hepatoma cell effects may be the mechanism for the development of Phyllanthus an effective anti-cancer drugs and lay the foundation.Methods:Using flow cytometry TUNEL assay Phyllanthus extract BEL-7404cells induced apoptosis; using microscopy cultured BEL-7404cells Phyllanthus extract morphological changes after treatment; through the flow PI-type cells were detected Phyllanthus extract on BEL-7404cell destruction; application flow cytometry CasPase-3activation level; immunochemical detection using cell Phyllanthus extracts treated human hepatoma BEL-7404Some cells are closely related proteins and apoptosis expression.Results:(1) Streaming TUNEL assay300μg/mL,500μg/mL Phyllanthus extract acts on BEL-7404, increased cell apoptosis rate (22.1±9.4)%,(21.6±8.3)%.(2) Microscope show that,when the concentration higher than300ug/mL the Phyllanthus extracts treated BEL-7404cell cytoplasm opacity, more cells were round, smaller in size, affixed to the wall is weakened, the refractive index variation, cellular debris, suspended and shedding of cells appeared in large numbers, and the cells spread clonal growth state.(3)300μg/mL-1000μg/mL of Phyllanthus extracts on BEL-7404cell cytotoxicity and its inhibition in a dose dependent manner.(4) Study results show that, CasPase-3of BEL-7404cells activity as Phyllanthus extract concentration increased significantly activated.(5) Immunocytochemistry showed, after Phyllanthus extract treating BEL-7404, Bcl-2protein expression was decreased, Bax, and Apaf-1protein expression increased.Conclusion:(1) Phyllanthus extract on BEL-7404cells have a more significant cytotoxicity, and the ability to induce apoptosis.(2) Phyllanthus extract can activate CasPase-3, enhanced Bax and Apaf-1protein expression, decreased Bcl-2protein expression, thereby inducing apoptosis. (3) Phyllanthus has been further developed as potential anti-hepatoma drugs.