| The research and exploitation of differ revulsant is a pop topic in the field of the anticancer drug's research .The differentiation and inducement of tumour is a phenomenon that malignant tumour cells differ to normal cells over again in the effects of differ revulsant. The most commonly used agent is retinoic acid, which is being used in leukaemia. However, most differentiation revulsants have toxicity to normal cells, and bring drug-resist to the cells for long-term. So we are searching the new differ revulsants which are more effective and fewer toxic, and effect in other targets.Sodium 4-phenylbutyrate (NaPB) is a new differ revulsant. It has short half-life, better penetrability, and has almost no toxicity tonormal cells. The known studies show that Sodium 4-phenylbutyrate has broad-spectrum antineoplastic effect. It can restrain not only the common leukaemia cells' growth extra-body, but also the prostate cancer and melanosarcoma in diferent degree. Because it can pass the blood-brain barrier easiely, we inverstigate the antineoplastic effect of Sodium 4-phenylbutyrate in human glioblastoma cell line SHG-44, and discuss its mechanism.Objective:1. Observe the effect of Sodium 4-phenylbutyrate on inducing growth inhibition and differentiation of cells' morphological in human glioblastoma cell line SHG-44.2. Assess the activity of Sodium 4-phenylbutyrate in combination with clinically active agents for encephaloma such as: VM26, cisplatinum(DDP), carmustine(BCNU) and common anticancer-drug adriamycin(ADM).Materials and methods:Cell culture. The human glioblastoma cell line SHG-44(obtained fromShanghai Medicine Technical Acedeme) was maintained in RPMI 1640supplemented with 10% fetal calf serum(FCS) at 37癈 in a 5% CO2atmosphere.Growth inhibition. The human glioblastoma cell line SHG-44 was plateduniformly in 96-well plates (Ixl0s cells/well) and allowed to adhereovernight. NaPB was formulated in RPMI 1640 media with 10% PCS and was added to the plates to obtain final concentrations of 125 ug/ml, 250 Ug/ml,500 ug/ml,1000 ug/ml,2000 ug/nd and 4000 ug/mL After 96 h of drug exposure, cell was added MTT for 4 h. abandoned the liquid and added the liquid (10%SDS-5%-0.012mmol/mlHCL), after half an hour, determined ASTO, then calculated the cell livability or cell growth inhibitory rate. Each experiment was performed in 6 wells and repeated thrice.Exposure time. To study the effect of exposure time of NaPB on growth inhibition, the cell line SHG-44 was plated uniformly in 96-well plates (1*105 cells/well) and allowed to adhere overnight. NaPB was added to the plates to obtain final concentration of 125yg4l, 250 yg/ml.SOO u g/ml,1000ng/ni],2000ug/ml and 4000 ng/mL After exposures of 48h, 72h and 96h,cell was added MTT for 4 h. abandoned the liquid and added the liquid (10%SDS-5%-0.012mmol/mlHCL), after half an hour, determined Aj7o, then calculated the cell livability or cell growth inhibitory rate. Each experiment was performed in 6 wells and repeated thrice. Morphologic differentation The glioblastoma cell line SHG-44 was plated on small culture-flasks (1><105 cells/well) and allowed to adhere overnight They were then exposed to NaPB at concentration of 500 pg/ml and 1 000 ug/ml for 72h. Then observe the cell shape and take photo. Growth inhibition of NaPB combined with VM26, cisplatinum, carmustine and adriamycin. Human glioblastoma cell line SHG-44 was uniformly plated in 96 well plates (1*105 cells/well) and allowed toadhere overnight. Then they were devided to two part: one was added NaPB (125ug/ml, 250iig/ml,500u&/iiil), after 4h, added VM26, cisplatinum, carmustine and adriamycin each; the other was added VM26, cisplatinum, carmustine and adriamycin each, after 4h, added NaPB (125 Wg/ml, 250 u g/inl,500 lig/ml). Cultured for 72h, then cell was added MTT for 4 h. abandoned the liquid and added the liquid(IO%SDS-5%-0.012mmol/mlHCL...
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