Effects Of Genistein On Expression Of Livin And Apoptosis, Cycle And Proliferation In Human Malignant Melanoma Libr Cells

Objective Clarify Genistein on the cells of malignant melanoma LiBr Livin geneexpression and apoptosis of malignant melanoma cell cycle and proliferation, and to provideexperimental basis to further explore the possibility of Genistein treatment of malignantmelanoma and reveals Genistein mechanism of tumor suppression.Methods Both at Gene and protein level, RT-PCR and Western blot are respectivelyused to confirm the delivery of Livin gene in malignant melanoma LiBr cells. After Genisteinis used, we analyze the change in the delivery level of Livin gene as well as dosage and timethrough RT-PCR. At cellular level, apply Annexin V-FITC/PI double-staining flow cytometryto detect cell apoptosis rate; at the protein level, apply Western blot to detect the expressionchange of Caspase-three enzymes and activation clips; use PI Single-staining flow cytometry todetect the LiBr cell cycle change, after the application of Genistein; use MTT colorimetricmethod to detect the LiBr cell proliferation change after the application of Genistein.Results Genistein could effectively knock down the Livin expression at mRNA in doseand time dependent manners,40umol/L of which achieved the highest knockdown effect onmRNA at72h after using Genistein. Genistein can induce the apoptosis of LiBr effectively,both in the early and late phases. LiBr cells in phase G0/G1increases notably, in contrast to thedecrease of those in phase-S, which demonstrates that cell generation cycle is prohibited.Genistein suppress LiBr generation in the range of48,72,96hours.Conclusions Expression of livin in LiBr cells could be efficiently knocked down byusing Genistein. induct LiBr cell apoptosis, restrain cell proliferation.