Effect Of Ultrasound Assistant ADM Chemotherapy On Breast Cancer

PART ONE Detecting oxygen species arising from the activation of ADM by ultrasonicObjectives:To investigate whether ultrasound can activate ADM and produce oxygen species without change the spectral characteristics of ADM.Methods:Using ROS probe to explore the wavelength intensity of ADM, and generation of reactive oxygen species, and using fluorescence staining method to explore the level of intracellular reactive oxygen species of cells. In addition, by analyzing the absorption spectrum curve of ADM to see the change of the molecular structure of ADM.Results:There are no difference in the fluorescence spectra curve between the ultrasound before and the ultrasound after. The the intervention of ultrasound after, the ADM solution make the curve peak increased, but after the flowing test, adding SOD, and NaN3 reduced the peak; ultrasound intervention of MCF-7/ADM cells and MCF-7/S cells DCF mean fluorescence intensity were 28.4±6.5,36.7±4.2,which were significantly lower than the corresponding ultrasound before the intervention (16.5±1.8,19.3±2.9) (P <0.05).Conclusions:After the above three experiments, we found that a certain wavelength intensity ultrasound can produce singlet oxygen 1O2 and the superoxide anion radical O2—(Experiment 2), and increase the level of ROS in breast cancer cells (experiment 3), without changing the molecular structure of ADM (Experiment 1).PART TWO Detection of Ultrasound-assistant ADM chemotherapy and the reversion of drug resistanceObjective:To study the effection of Ultrasound (US) assistant ADM therapy and the reversion of drug resistanceMethods:A human breast cancer cells MCF-7/S and adriamycin-resistant human breast cancer cells MCF-7/ADM were on this study. By cell growth curve, detection MCF-7/ADM resistance protein MDR1, P-gP expression, measure the concentration of doxorubicin in breast cancer cells, test the apoptosis of cells and value the reversion of drug resistance and mechanisms of US assistant ADM therapy.Results:"US+ADM" on MCF-7 cells were inhibited and MCF-7/ADM; the drug concentration increased, the inhibitory effect is investigated. After the ultrasound intervention,the IC50 of MCF-7/ADM cells was 12.96, the drug resistance reversation was 3.76 multiple. Doxorubicin within cells of Ultrasound treated MCF-7/S and MCF-7/ADM were significantly increased. RT-PCR analysis showed that:MDR1 mRNA, MRP1 mRNA expression of "US+ADM" group were significantly lower than ADM alone group and the control grou(P<0.05). On "ultrasound+ADM" treatment group,the P-gp protein, MRP1 protein expression were significantly lower than those of ADM group and the control group(P<0.05). AO/EB staining showed cell morphology:ultrasound after the intervention significantly increased apoptosis, compared with the control group (P<0.05).Conclusion:"US+ADM" could inhibit resistant cell surface markers P-gp, MRP expression on adriamycin resistant human breast cancer cell lines. And increase doxorubicin concentration of MCF-7/ADM cells and sensitive strains of MCF-7 cells. Ultrasound could inhibit the cell proliferation and reverse the drug resistance of ADM. PART THREE Effect of ultrasound assistant ADM chemotherapy on the mouse mammary cancerObjective:To observe the ultrasound assistant ADM therapy on the mouse mammary cancerMethods:BALB/c mice were injected subcutaneously in the right axillary MA782 cells of mouse breast cancer, and a total of 40 mice was randomly divided into 4 groups: simple ultrasound group, ultrasound+ADM group, simple ADM group and control group; tumor growth curves and the survival curves of mice were drawn to explore the effect of ultrasound assistant ADM chemotherapy on the mouse mammary cancer.Results:The "US+ADM" group of mice tumor growth was significantly inhibited, compared with ADM alone, There was significant difference (p<0.01), while the ultrasound-assisted ADM group overall survival time of mice, compared with ADM alone had significant differences (p<0.05). ADM alone group showed some anti-tumor effects, tumor growth slow, and simple inhibition of ultrasonic little intervention.Conclusion:Tumor growth of "US+ADM" group were inhibited, the survival time of the mouse of mammary cancer were prolonged.