The Study Of Ultrasound Destruction LHRH Targeted Microbubbles Combined Paclitaxel Treatment Ovarian Cancer

PARTⅠPREPARATION AND OBSERVATION OF TARGETED ABILITY OF LIPOSOME MICROBUBBLES TARGETED HUMAN OVARIAN CANCER CELL LINE OVCAR-3 IN VITROObjective: To prepare human ovarian cancer—targeted liposome microbubbles and assess its targeted ability in vitro.Methods: ligand LHRH was attached to the surface of self-made liposome microbubbles by covalent bonding to prepare targeted microbubbles. Immunofluorescent staining assay Was used to identify the combination of LHRH with liposome microbubbles and the targeted microbubbles were added to ovarian cancer cells ovcar-3 and then observed under the light and fluorescence microscope to evaluate the targeting ability of the targeted paclitaxel liposome microbubbles with ovarian cancer cells in vitro ,while the common liposome microbubbles were controls.And use LHRH to do a block experiment.Results:Targeted microbubbles were positive in immunofluorescent straining assay . In vitro study of the targeting ability showed the targeted microbubbles could actively adhere to ovcar-3 cel1. While the control Was negative. And LHRH can effectively block the targeted microbubbles .Conclusion: The targeted liposome microbubbles can be prepared successfltlly by covalent bonding method ,this targeted microbubbles could bind to human ovarian cancer cells specially and efectively in vitro.PARTⅡULTRASOUND DESTRUCT LHRH TARGETED LIPOSOME MICROBUBBLES COMBINED PACLITAXEL ON OVARIAN CARCINOMA CELL PROLIFERATION AND APOPTOSIS AND INTRACELLULAR DRUG CONCENTRATIONobjective: To study the effects of ultrasound combined targeted liposome microbubbles(TLM) synergies paclitaxel treatment on Ovcar3 cell proliferation, apoptosis and intracellular drug concentration(IDC).Methods: The concentration of paclitaxel treatement on Ovcar3 cell in vitro was determined by MTT assay. Ovcar3 cell proliferation situation was also determined by MTT assay after ultrasound combined TLM synergies paclitaxel treatment. Ovcar3 cell apoptosis was detected by flow cytometry(FCM) . Intracellular drug concentration was observed by high performance liquid chromatogram HPLC.Results: The concentration of paclitaxel treatement on Ovcar3 cell in vitro was 10-6mol/L(P﹤0.050).Ultrasound combined paclitaxel, paclitaxel alone, ultrasound combined LM synergies paclitaxel, ultrasound combined TUCA synergies paclitaxel and all had significant inhibition effects on OVCAR3 cell proliferation. However, among the four groups, the effects of ultrasound combined TUCA synergies paclitaxel group was the strongest(P﹤0.010) . And TUCA alone had no significant effects on cells. The apoptosis rates of cells determined by FCM were respectively (1.68±0.14)% in the contrast group, (6.52±0.23)% in the TUCA alone group, (11.32±2.57)% in paclitaxel alone group, (15.87±0.65)% in the ultrasound combined paclitaxel group and (21.40±0.56)% in the ultrasound combined LM synergies paclitaxel group and (35.27±0.62)% in the ultrasound combined TUCA synergies paclitaxel . The intracellular drug concentration of OVCAR3 cell was up regulated significantly among all the groups.Conclusion: Ultrasound combined TUCA synergies paclitaxel method could not only inhibit SKOV3 cell proliferation, but also induce apoptosis and up regulate intracellular drug concentration.