Preparation Of Anti-STEAP-1 Nano-microbubbles Targeting Prostate Cancer PC3 Cells And Its Targeting Study

Part 1 Preparation of Anti-STEAP-1 Nano-microbubbles Targeting Prostate Cancer PC3 Cells and Its Targeting Study in Vitro Objective: To prepare nano-microbubbles specifically targeting prostate cancer PC3 cells,characterize their physical and chemical properties and verify their targeting ability in vitro.Methods: Rotating evaporation method,mechanical vibration method were used to prepared non-targeting nano-microbubble and the biotinylated one.By using the biotinavidin method,conjugate biotinylated STEAP-1 antibodies to biotinylated nanomircobubbles generated biotinylated anti-STEAP-1 nano-microbubble targeting specifically prostate cancer PC3 cells,and then were observed via immunofluorescence staining.Observe and detect the morphology,distribution and particle size of microbubbles,and the change of particle size during one week.The in vitro targeting ability to prostate cancer PC3 cells was observed with inverted microscope.Results: The non-targeted nano-microbubbles and the targeted one were successfully generated.They were round,uniformly distributed and free of aggregation.After beingkept at the temperature of 4 degrees for one week,no significant difference of the two nano-microbubbles' partical size was detected.In in vitro targeting experiments,the targeted nano-microbubbles attached to the surface of PC3 cells,and the surface of PC3 cells previously treated with biotinylated STEAP-1 antibody had no aggregation of the targeting nano-microbubbles.Conclusion: The biontinylated anti-STEAP-1 nano-microbubbles specifically targeting prostate cancer PC3 cells can be prepared successfully through rotating evaporation method,mechanical vibration method and biotin-avidin method.And they can specifically bind to prostate cancer PC3 cells in vitro.Part 2 Experimental Study of Self-made Anti-STEAP-1 Nano-microbubbles Ultrasound Contrast Agents Targeting Tumor-bearing Mice Objective: To investigate the ultrasound imaging characteristic of the self-made antiSTEAP-1 nano-microbubbles in nude mice model bearing PC3 prostate carcinoma.Methods: Rotating evaporation method,mechanical vibration method were used to prepared non-targeting nano-microbubble and the biotinylated one.By using the biotinavidin method,conjugated biotinylated STEAP-1 antibodies to biotinylated nanomircobubbles generated biotinylated anti-STEAP-1 nano-microbubble targeting specifically prostate cancer PC3 cells.To establish a nude mouse xenograft model of prostate carcinoma PC3 cells,respectively,using two kinds of contrast agent to perform contrast-enhanced ultrasoud on nude mice xenograft.To play attention to the perfusion and distribution characteristic,and keep the video data down.To analysis the results of Peak Intensity(PI),Time to Peak(TP),Area under the Curve(AUC),and Sharpness By using Qontra Xt dynamic contrast quantitative analysis software.Results: The non-targeting nano-microbubbles and the targeting one were successfully generated.The prostate cancer PC3 xenograft model was successfully established.The enhanced imaging results in vivo showed that the anti-STEAP-1 targeted nanomicrobubbles ultrasound contrast agent was slightly shorter than the non-targeted nanomicrobubbles,but its enhanced development effect was obvious.Conclusion: The biontinylated anti-STEAP-1 nano-microbubbles specifically targeting prostate cancer PC3 cells can be preparedsuccessfully through rotating evaporation method,mechanical vibration method and biotin-avidin method.And they can specifically bind to prostate cancer PC3 cells in vivo,which can enhance development effect and may be helpful for tumor diagnosis.