Study Of LHRHa-Targeted Microbubble Contrast Agent Assisted With Ultrasound Combined Cisplatin On Apoptosis Of Ovarian Cancer Cell A2780/DDP

Object:to study the effect of Ultrasound irradiation LHRHa targeted ultrasound contrast agent combined with Cisplatin treatment on A2780/DDP cell proliferation and apoptosis.Method:MTT assay was used to test the cell viability of cell line A2780 and A2780/DDP 24 hours after treatment in different concentration of DDP, SPSS software was used to anlysis the 50% inhibiting concentration of A2780 and A2780/DDP cell, then to get the drug resistance index of cell line A2780/DDP. The human ovarian cancer cell line A2780/DDP was divided into6 groups: control group, liposome microbubble(LM) plus ultrasound group, DDP group, DDP and ultrasound radiation group, DDP and LM plus ultrasound radiation group, and DDP and LHRHa-bound liposome microbubles (LHRHa-LM) plus ultrasound radiation group. MTT assay was used to test for cell proliferation, flow cytometry for cell apoptosis and cell cycle, laser confocal microscopy for expressing of bax/bcl-2, transmission election microscopy for morphology.Result:the drug resistance index of cell line A2780/DDP was 2.45. In DDP and LHRHa-bound liposome microbubles (LHRHa-LM) plus ultrasound radiation group, cell proliferation was obviously inhibited (p<0.05), the CPIR was (44.21±3.05) %, and apoptosis ratio was( 26.25±1.85)%,evidently higher than other groups(p<0.05).And S/G2 was remarkably up-regulated(p<0.05),so was the index of bax/bcl-2.Conclusion : Ultrasound irradiation LHRHa targeted ultrasound contrast agent combined with Cisplatin had obviously effect on cell proliferation inhibition and apoptosis induction of ovarian cancer cell line A2780/DDP.PartⅡMechanism of ultrasound mediated LHRHa Carrying microbubbles destruction combined with Cisplatin (DDP) in apoptosis induction of ovarian cancer cell A2780/DDPObjective:to investigate the effects of the ultrasound combine with LHRHa-bound liposome microbuble s (LHRHa-L M) in intracellur Cisplatin (DDP) concentration.Method:FCM was used to test PI immediately, 12h, 24h, 48h after microbubble combined ultrasound irradiation.The human ovarian cancer cell line A2780/DDP was divided into 5 groups: control group, DDP group, DDP and ultrasound radiation group, DDP and LM plus ultrasound radiation group, and DDP and LHRHa-LM plus ultrasound radiation group. High Performance Liquid Chromatography ( HPLC )was used to detect intracellular drug concentration at 12h, 24h, 48h after treatment for every group. F low Cytometry ( FCM ) test was used to detect the cytomembrane permeability to propidine iodide(PI) instantly, 12h, 24h after liposome microbubles (LM )combined ultrasound irradiation in vitro A2780/DDP cell.Result:cytomembrane permeability to PI has increased instantly after treatment, and decreaced over time. DDP concentration is in linear dependence, R is 0.9994. DDP and LHRHa-LM plus ultrasound group demonstrated the most obvious increasing of intracellular drug concentration on 12h, 24h after treatment.Conclusion:LHRHa-LM plus ultrasound could increase permeability of cell membrane more markedly than other groups, so to increase the intracellular chemotherapy drug concentration, and the rate of drug increasing may slower over time, that may be related to reversible increasing of cytomembrane permeability caused by LM plus ultrasound.