【摘要】：研究背景：膀胱腫瘤（Bladder carcinoma）是臨床上最常見的惡性腫瘤之，其發(fā)病率和死亡率在泌尿系腫瘤中均居首位。非肌肉浸潤性膀胱癌(non-muscleinvasive bladder cancer）約占全部膀胱腫瘤的75—85%，雖然其惡性程度低，但大約有10%的患者最終可發(fā)展為肌肉浸潤性膀胱癌。因此如何對膀胱腫瘤進行早期診斷和靶向治療就成為當今醫(yī)學領域研究的熱點內容。 目的：建立中國人膀胱癌BIU-87細胞系的裸鼠皮下移植瘤模型，，對小分子環(huán)七肽片段-SISSLTH在細胞和組織不同水平進行驗證，鑒定該導向肽對中國人膀胱癌BIU-87細胞結合的特異性和親和性，為未來臨床上對膀胱腫瘤進行早期診斷、術中準確切除和開發(fā)靶向藥物提供定的理論基礎。 方法：將Matrigel基質膠作為中國人膀胱癌BIU-87細胞的懸浮劑，用注射器將癌細胞種植于裸鼠腋窩背側皮下建立裸鼠移植瘤模型。通過化學方法合成小分子環(huán)七肽片段-SISSLTH，純化并標記綠色熒光染料異硫氰酸熒光素FITC，利用免疫熒光技術原理，通過應用流式細胞術和激光共聚焦顯微鏡技術兩種方法，對小分子多肽熒光探針FITC-SISSLTH在細胞和組織不同水平進行驗證，鑒定其與中國人膀胱癌細胞結合的特異性和親和性，以及該小分子探針在荷瘤裸鼠體內的生物分布情況。 結果：在細胞水平對小分子多肽熒光探針FITC-SISSLTH進行驗證，其結果顯示：小分子多肽熒光探針FITC-SISSLTH與中國人膀胱癌BIU-87細胞系的親和性明顯高于人肝癌Hep-G2細胞系和人前列腺癌PC-3細胞系。利用激光共聚焦顯微鏡技術研究小分子探針在荷瘤裸鼠體內的分布，實驗結果表明：該探針可以特異性地在腫瘤組織中富集，而其他臟器中聚集很少。說明該探針對膀胱癌組織有特異性和靶向性。 結論：在細胞水平對小分子多肽熒光探針FITC-SISSLTH進行驗證，其結果顯示：小分子多肽熒光探針FITC-SISSLTH與中國人膀胱癌BIU-87細胞系的親和性明顯高于人肝癌Hep-G2細胞系和人前列腺癌PC-3細胞系。利用激光共聚焦顯微鏡技術研究小分子探針在荷瘤裸鼠體內的分布，實驗結果表明：該探針可以特異性地在腫瘤組織中富集，而其他臟器中聚集很少。說明該探針對膀胱癌組織有特異性和靶向性。
[Abstract]:Background: bladder tumor (Bladder carcinoma) is one of the most common malignant tumors in clinic. Non-muscle invasive bladder cancer (non-muscleinvasive bladder cancer) accounts for 75-85% of all bladder tumors. Although its malignancy is low, about 10% of the patients can eventually develop musculoinvasive bladder cancer. Therefore, how to make early diagnosis and targeted therapy of bladder tumors has become a hot topic in the field of medicine. Objective: to establish a subcutaneous transplanted tumor model of Chinese bladder cancer BIU-87 cell line in nude mice and to verify the different levels of small molecule cyclic heptapeptide fragment (SISSLTH) in cells and tissues. The specificity and affinity of the targeting peptide to BIU-87 cells in Chinese bladder cancer were identified, which provided a theoretical basis for early diagnosis, intraoperative excision and development of targeted drugs for bladder cancer in the future. Methods: Matrigel matrix glue was used as the suspension of Chinese bladder cancer BIU-87 cells. The tumor cells were implanted into the dorsal subcutaneous axillary of nude mice with syringe to establish the transplanted tumor model in nude mice. SISSLTH, was synthesized by chemical method for the purification and labelling of green fluorescent dye fluorescein isothiocyanate (FITC,). Flow cytometry and laser confocal microscopy were used to purify and label the green fluorescent dye fluorescein isothiocyanate (FITC,) by using the principle of immunofluorescence. The specificity and affinity of the small molecular polypeptide fluorescence probe (FITC-SISSLTH) to Chinese bladder cancer cells were confirmed at different levels of cell and tissue, and the biological distribution of the small molecule probe in nude mice was also studied. Results: the fluorescence probe FITC-SISSLTH of small polypeptide was verified at the cell level. The results showed that the affinity of small molecular peptide fluorescent probe FITC-SISSLTH to Chinese bladder cancer BIU-87 cell line was significantly higher than that of human hepatocellular carcinoma Hep-G2 cell line and human prostate cancer PC-3 cell line. The distribution of small molecular probe in nude mice bearing tumor was studied by laser confocal microscopy. The experimental results showed that the probe could be specifically enriched in tumor tissue, but few in other organs. The results showed that the probe was specific and targeted to bladder cancer. Conclusion: the fluorescence probe FITC-SISSLTH of small polypeptide was verified at cell level. The results showed that the affinity of small molecular peptide fluorescent probe FITC-SISSLTH to Chinese bladder cancer BIU-87 cell line was significantly higher than that of human hepatocellular carcinoma Hep-G2 cell line and human prostate cancer PC-3 cell line. The distribution of small molecular probe in nude mice bearing tumor was studied by laser confocal microscopy. The experimental results showed that the probe could be specifically enriched in tumor tissue, but few in other organs. The results showed that the probe was specific and targeted to bladder cancer.
【學位授予單位】：山西醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R737.14

【參考文獻】

相關期刊論文 前10條

1 俞莉章,黃雅麗,楊晨,宋蓮芬,徐令巽,顧方六;人體膀胱移行細胞癌細胞系BIU-87的建立及其生物學特性[J];北京醫(yī)科大學學報;1989年01期

2 張龍江,祁吉;分子影像學中的一些常用術語[J];放射學實踐;2005年04期

3 周非凡;宋盛;吳寶艷;歐忠敏;邢達;;靶向納米探針介導的近紅外激光熱療[J];激光生物學報;2011年01期

4 鄧大偉;劉飛;曹潔;陳新洋;顧月清;;兩種近紅外熒光探針的合成及腫瘤靶向研究[J];中國激光;2010年11期

5 雷霞;伍津津;;基質膠的生物學活性研究進展[J];口腔醫(yī)學;2011年06期

6 李俊;楊曉峰;劉仍新;;小分子肽CSNRDARRC與膀胱移行細胞癌組織特異性結合的研究[J];現(xiàn)代泌尿外科雜志;2012年03期

7 趙揚;楊曉峰;郝小強;;多肽CSNRDARRC與膀胱癌細胞結合親和性及其位點的研究[J];現(xiàn)代泌尿生殖腫瘤雜志;2011年02期

8 陳興屹;溫機靈;陳衛(wèi)華;溫曉飛;楊波;王躍閩;;高危非肌層浸潤性膀胱癌二次經(jīng)尿道電切治療體會[J];同濟大學學報(醫(yī)學版);2011年01期

9 張qZ;周虎臣;;小分子探針在確定活性化合物的生物靶點中的應用[J];藥學學報;2012年03期

10 韓月東;崔大祥;宦怡;李智銘;劉賀亮;宋華;劉彬;杜彤;高峰;賀蓉;;熒光磁性納米粒子與PSA單鏈抗體復合探針對前列腺癌的靶向顯像和治療[J];中國腫瘤生物治療雜志;2008年05期

本文編號：2351858