The Experimental Study Of Docetaxel On Human Hepatoma SMMC-7721Cells Toxicity And Radiosensitization In Vitro

Objective:To study of docetaxel on human hepatoma SMMC-7721cells toxicity and radiosensitization, to provide experimental evidence for the treatment of liver cancer.Materials and Methods:(1) The recovery of human hepatoma cell line SMMC-7721were cultured and passaged to take experimental logarithmic growth phase cells.(2) MTT colorimetric determination of inhibition of docetaxel growth of liver cancer cells.(3) To observe the optical inverted microscope morphological changes on docetaxel-treated cells after different times.(4) To analyze the effects on cell cycle distribution and apoptosis using flow cytometry.(5) colony formation was applied to observe medicine with radiotherapy sensitization effect on cells.(6) To detect the changes of intracellular reactive oxygen species by flow cytometry after the role of drugs and gamma-ray.Results:(1) Human hepatoma cell SMMC-7721by the recovery of culture, passaged3-5days, the cells grew well, and entered the logarithmic growth phase.(2) MTT colorimetric assay results showed that:docetaxel inhibited the proliferation of human hepatoma SMMC-7721cells, and showed a drug concentration-dependent drug50%inhibitory concentration (IC50)2.199nmol/ml, IC10for0.439nmol/ml.(3) Optical inverted microscope visible:cells with IC10(0.439nmol/ml) drug concentration effect at different times, with the extension of the time of drug action. light microscope, the continuity of change visible cell damage, cell gap widened, polymorphic adherent cells, adherent state variation, to the formation of intracellular vacuoles. not adherent growth, eventually apoptosis with large amounts of cavity loss.(4) Flow cytometry for cell cycle distribution and apoptosis measurement results show:the concentration of cells were (0.005nmol/ml) was significantly lower than docetaxel role when IC10(0.439nmol/ml) cells for24hours, the cell cycle G2/M phase arrest, for48hours after induction of apoptosis, and24hours the drug concentration50nmol/ml role to induce apoptosis, showing a drug concentration-the role of time-dependent manner.(5) set off the formation of experiments the results show:docetaxel on human hepatoma cell radiosensitizing effect, medication alone, simple irradiation, Joint group1combined group2cell survival scores were decreased, and showing the drug concentration-irradiation dose-dependent; joint group on cell survival fraction is particularly significant with medication alone or radiation alone, there were statistically significant differences (P<0.01). due to biological effects than medication alone and radiation alone sum of radiosensitization ratio SER is greater than1, which combined group1was1.03, the joint group2was1.26.(6) the measurement results of flow cytometry of intracellular reactive oxygen species (ROS):medication alone group, irradiation group, the combined group of intracellular reactive oxygen water were significantly increased, and showed a dose-dependent effect, which combined group activity oxygen levels increased more obvious, and irradiation group and the treatment group alone were significantly different (P<0.01), and due to the biological effect is greater than the sum of the medication alone group and irradiation group.Conclusion:Docetaxel has toxic effect and radiotherapy sensitization on human hepatocellular carcinoma cells SMMC-7721, which may be caused cells in G2/M phase arrest (radiosensitization phase) related to, and in sensitizing under the conditions of intracellular reactive oxygen species levels were significantly increased, may enhance cell radiosensitivity and apoptosis-inducing aspects also played a certain role in promoting.