Experimental Studies On Paclitaxel-induced Apoptosis And Growth Inhibitory Effects In Ewing's Sarcoma Cell Line Of RD-ES In Vitro

Objective To investigate the apoptosis and growth inhibitory effects of paclitaxel on Ewing's sarcoma cell line RD-ES and to offer laboratorial foundation of experiment of using paclitaxel in vivo and clinical application of paclitaxel.Methods1. The RD-ES cells were resuscitated, cultivated, passaged and propagated in mediums, with different concentration of Paclitaxel,Cisplatin(DDP), Adriamycin(ADM)for 0, 8, 16, 24, 48, 72 hours at Logarithm growth phase. RD-ES cells in drug-free medium saved as control. Viable cells were counted by trypan blue dye exclusion and the rates of cell growth inhibition were observed.2. The cultivated RD-ES cells were planted to flasks containing flakes and treated with different concentration of chemotherapeutic drugs for different times as above. Giemsa' stain were used to observe the cells' shape.3. The cell cycle and apoptosis rates of the above cells were determined by flow cytometry(FCM).4. The apoptosis of the cells at Logarithm growth phase were also observed by TdT mediated dUDP nick end labeling (TUNED,apoptosis cells were counted.Results Compared with other drugs, paclitaxel has obvious effects on inducing apoptosis and growth inhibitory which depend on time or dose increasing, with rate of viable cells decreasing, the number of apoptosis cells increasing, cells deforming, an apoptosis peak in FCM being evident and DNA cleavage being lebeled in positive reaction for TUNEL .Conclusion Paclitaxel can inhibit the mitosis and induce apoptosis of Ewing's sarcoma cell line of RD-ES in a time- and dose-dependented manner in vitro. The resulst of this study can serve as a base of clinical trail in vivo.