本文選題：NGR-釓 + 腫瘤細胞��； 參考：《遵義醫(yī)學院》2014年碩士論文

【摘要】：目的：CD13是腫瘤細胞膜及腫瘤新生血管內(nèi)皮細胞膜上的受體，對腫瘤細胞的生長、繁殖、侵襲及轉(zhuǎn)移起著重要作用。本研究以腫瘤細胞膜上CD13受體為靶點，合成能與其特異性結合的配體NGR短肽，并以釓及熒光作為MR的成像部分，構建熒光-NGR-釓的新型多模態(tài)MR靶向?qū)Ρ葎⑻接懺揗R對比劑在體外的生物活性及其體外靶向性能，為活體內(nèi)靶向成像奠定實驗基礎。 方法：通過免疫組化實驗檢驗腫瘤細胞膜上CD13表達情況。分為實驗組（HePG2人肝癌細胞和HT1080人肉瘤細胞）和對照組（HT29人結腸癌細胞）.用化學合成方法偶聯(lián)對比劑。首先合成熒光-NGR，再合成熒光-NGR-釓對比劑，并進行相關理化數(shù)據(jù)測試最后細胞熒光實驗驗證熒光-NGR體外靶向性能：a.實驗組：實驗組（HePG2，HT1080細胞）和對照組(HT29細胞)爬片上加等量熒光-NGR，觀察細胞爬片熒光強度。b.通過拮抗實驗進一步從反面驗證NGR對CD13的體外靶向性。分為兩批實驗：第一批HT1080細胞拮抗組，提前在HT1080腫瘤細胞爬片上滴加非熒光NGR，封閉CD13靶點之后再滴加與同一次實驗組等劑量熒光-NGR，觀察每組細胞爬片熒光強度；第二批HePG2拮抗組，提前在HePG2腫瘤細胞爬片上滴加非熒光NGR，之后再滴加熒光-NGR，劑量和實驗組（同一次實驗）是一致的，觀察每組細胞爬片熒光強度，及細胞熒光實驗驗證熒光-NGR-釓對比劑體外靶向性能。a.對比觀察熒光-NGR-釓對CD13高表達和CD13低表達細胞的結合能力：實驗組用HT1080細胞，對照組用HT29細胞，同時在HT1080和HT29細胞爬片上分別滴加熒光-NGR-釓，觀察兩種細胞爬片上的熒光強度。b.通過拮抗組實驗反證熒光-NGR-釓在體外的生物活性。實驗分為HT1080細胞實驗組和拮抗組，實驗組細胞爬片上分別滴加遞減劑量熒光-NGR-釓，拮抗組HT1080細胞爬片或腔式蓋玻片上事先滴加遞增劑量非熒光NGR，然后再加入與同次實驗組等量的熒光-NGR-釓劑量，比較實驗組和拮抗組熒光的強度。 結果：實驗組HePG2和HT1080細胞免疫組化實驗呈CD13高表達，對照組HT29細胞呈CD13低表達；.熒光-NGR與熒光-NGR-釓體外偶聯(lián)成功；實驗組HePG2和HT1080細胞熒光-NGR實驗均可看到大量熒光標記物，對照組僅看到微弱熒光標記物在HT29細胞上。實驗組熒光強度高于對照組及拮抗組，半定量對比分析，差異有顯著統(tǒng)計學意義（P0.005）；熒光-NGR-釓實驗實驗組（HT1080細胞）熒光強度高于對照組（HT29細胞）及拮抗組（HT1080細胞），半定量對比分析，差異有顯著統(tǒng)計學意義（P0.005）。 結論：熒光-NGR-釓偶聯(lián)成功，，在體外對HePG2和HT1080腫瘤細胞膜上的CD13具有特異性靶向性。
[Abstract]:Objective CD13 is a receptor on tumor cell membrane and tumor neovascularization endothelial cell membrane, which plays an important role in the growth, proliferation, invasion and metastasis of tumor cells. In this study, the ligand NGR short peptide, which can specifically bind to CD13 receptor on tumor cell membrane, was synthesized, and gadolinium and fluorescence were used as Mr imaging part to construct a novel multimode Mr targeting contrast agent with fluorescence-NGR- gadolinium. The bioactivity of Mr contrast agent in vitro and its in vitro targeting performance were discussed, which laid an experimental foundation for in vivo target imaging. Methods: the expression of CD13 on tumor cell membrane was detected by immunohistochemistry. Two groups were divided into two groups: the experimental group (HePG2 human hepatoma cell and HT1080 human tumor cell) and the control group (HT29 human colon cancer cell line). Coupling contrast agent by chemical synthesis method. Fluorescence-NGR was synthesized at first, then fluorescence-NGR-gadolinium contrast agent was synthesized. In the experimental group, the same amount of fluorescence (NGR) was added to the climbing slices of the experimental group (HePG2 + HT1080 cells) and the control group (the HT29 cells), and the fluorescence intensity of the cell climbing slices was observed. The in vitro targeting of NGR to CD13 was further verified by antagonistic experiments. In the first batch of HT1080 cell antagonistic group, non-fluorescent NGR was added on the HT1080 tumor cell climbing slice in advance. After blocking the target of CD13, the same dose of fluorescence was added to the same experimental group, and the fluorescence intensity of each group was observed. In the second group of HePG2 antagonists, non-fluorescent NGRs were added to the HePG2 tumor cells in advance, and then fluorescent NGR was added later. The dose was the same as that in the experimental group (the same experiment). The fluorescence intensity of the climbing slices of each group was observed. The in vitro targeting performance of fluorescent-NGR-gadolinium contrast agent was verified by cell fluorescence assay. The binding ability of fluorescent-NGR- gadolinium to high expression of CD13 and low expression of CD13 was observed: HT1080 cells were used in experimental group, HT29 cells were used in control group, and fluorescence-NGR- gadolinium was added to HT1080 and HT29 cell climbing slices respectively. The bioactivity of fluorescence-NGR-gadolinium in vitro was confirmed by antagonistic experiment. The experiment was divided into two groups: HT1080 cell experimental group and antagonistic group. In the antagonistic group, the fluorescence intensity of the experimental group and the antagonistic group were compared by adding the increasing dose of non-fluorescent NGR to the climbing slice or cavity glass slide, and then adding the same dose of fluorescence-NGR- gadolinium as the same experimental group. Results: the expression of CD13 was high in HePG2 and HT1080 cells in the experimental group, and the low expression of CD13 in the HT29 cells in the control group. HePG2 and HT1080 cells in the experimental group could see a large number of fluorescent labeled substances, while in the control group, only the weak fluorescent labeled substance could be seen on the HT29 cells. The fluorescence intensity of experimental group was higher than that of control group and antagonistic group. The fluorescence intensity of HT1080 cells in the fluorescent NGR- gadolinium group was higher than that in the control group (HT29 cells) and in the antagonist group. The semi-quantitative comparison analysis showed that the difference was statistically significant (P 0.005). Conclusion: fluorescence-NGR-gadolinium coupling is successful and has a specific targeting of CD13 on the cell membrane of HePG2 and HT1080 tumors in vitro.
【學位授予單位】：遵義醫(yī)學院
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R445.2;R73-3

【參考文獻】

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本文編號：1913072