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  本文選題：前列腺跨膜上皮1抗原　切入點：血管內(nèi)皮生長因子受體　出處：《皖南醫(yī)學院》2017年碩士論文　論文類型：學位論文

【摘要】：目的:使用前列腺跨膜上皮抗原-1(STEAP-1)抗體及血管內(nèi)皮生長因子受體2(VEGFR-2)抗體作為配體,采用生物素-親和素法制備攜帶上述抗體的新型靶向超聲微泡造影劑,并探討其體外尋靶能力,分析所制備造影劑在荷人前列腺癌裸鼠增強顯影效果,希望獲得有價值的早期超聲診斷方法。方法:(1)采用生物素-親和素方法制備靶向超聲造影劑,將STEAP-1抗體及VEGFR-2抗體與普通超聲造影劑進行結(jié)合,并在熒光顯微鏡下觀察微泡熒光情況,并通過流式細胞儀檢測其結(jié)合率及穩(wěn)定性。(2)在無菌環(huán)境下體外培養(yǎng)人前列腺癌細胞株P(guān)C3并傳代,在雄性裸鼠皮下接種PC3細胞,建立荷人前列腺癌移植瘤雄性裸鼠模型。通過荷瘤裸鼠模型尾靜脈注射靶向超聲微泡造影劑及普通超聲造影劑進行超聲增強顯影,觀察裸鼠移植瘤的造影劑分布特征及實時灌注情況,并用相關(guān)軟件繪制時間-強度曲線進行定量分析,比較不同造影劑的達峰時間(Time to Peak,TTP)、峰值強度(Peak Intensity,PI)及平均渡越時間(Mean Transit Time,MTT)。結(jié)果:(1)攜帶STEAP-1抗體及VEGFR-2抗體的靶向微泡超聲造影劑通過免疫熒光實驗在熒光顯微鏡下均呈陽性表達,即微泡邊緣出現(xiàn)綠色熒光。(2)成功建立荷人前列腺癌裸鼠模型,通過鼠尾靜脈注射普通超聲造影劑及兩種靶向造影劑,所有組別均可見明顯增強,達到一定峰值后逐漸減弱。結(jié)論:(1)采用生物素-親和素法可以成功制備攜帶STEAP-1抗體及VEGFR-2抗體的靶向超聲造影劑,且兩種靶向超聲造影劑性質(zhì)均較穩(wěn)定;(2)在體外實驗中,攜帶STEAP-1抗體及VEGFR-2抗體的靶向超聲造影劑在荷瘤裸鼠中靶向性較好且特異性較高,其中VEGFR-2抗體可以與前列腺癌新生血管內(nèi)皮細胞特異性結(jié)合,為評估前列腺癌新生血管形成提供了新的思路。
[Abstract]:Objective: to prepare a new type of ultrasound microbubble contrast agent carrying the antibodies against prostate transmembrane epithelial antigen-1 (STEAP-1) and vascular endothelial growth factor receptor 2VEGFR-2 (VEGFR-2) by biotin-avidin method. The aim of this study was to investigate the ability of target finding in vitro, to analyze the enhancement effect of the contrast agent in nude mice bearing human prostate cancer, and to obtain a valuable method for early ultrasound diagnosis. Methods: biotin-affinity method was used to prepare the target ultrasound contrast agent. STEAP-1 antibody and VEGFR-2 antibody were combined with conventional ultrasound contrast medium, and the fluorescence of microbubbles was observed under fluorescence microscope. The binding rate and stability of human prostate cancer cell line PC3 were detected by flow cytometry. PC3 cells were cultured and subcultured in vitro. PC3 cells were subcutaneously inoculated into male nude mice. The male nude mice model of transplanted tumor of human prostate cancer was established. Ultrasound enhanced imaging was performed by injection of targeted ultrasound microbubble contrast agent and conventional ultrasound contrast agent into tail vein of nude mice bearing human prostate cancer. The distribution characteristics of contrast media and the real-time perfusion of transplanted tumor in nude mice were observed and the time-intensity curves were plotted by relevant software for quantitative analysis. The peak time and peak intensity of different contrast agents were compared. The peak intensity and the mean transit time were compared. The results showed that the microbubble ultrasound contrast agents carrying STEAP-1 antibody and VEGFR-2 antibody were all positive under fluorescence microscope. That is, green fluorescence appeared at the edge of the microbubble. (2) A nude mouse model of human prostate cancer was successfully established. The normal ultrasound contrast agent and two target contrast agents were injected into the tail vein of the mouse, and the enhancement was observed in all groups. Conclusion: in vitro, STEAP-1 antibody and VEGFR-2 antibody targeting ultrasound contrast agent can be successfully prepared by using biotin-avidin method, and the properties of both targeted ultrasound contrast agents are stable in vitro. The ultrasound contrast agent carrying STEAP-1 antibody and VEGFR-2 antibody was more specific and specific in tumor-bearing nude mice. VEGFR-2 antibody could specifically bind to prostate cancer neovascularization endothelial cells. It provides a new idea for evaluating the angiogenesis of prostate cancer.
【學位授予單位】：皖南醫(yī)學院
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R445.1;R737.25

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