【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(rùn)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(rùn)性膀胱癌。因此如何對(duì)膀胱腫瘤進(jìn)行早期診斷和靶向治療就成為當(dāng)今醫(yī)學(xué)領(lǐng)域研究的熱點(diǎn)內(nèi)容。 目的：建立中國(guó)人膀胱癌BIU-87細(xì)胞系的裸鼠皮下移植瘤模型，，對(duì)小分子環(huán)七肽片段-SISSLTH在細(xì)胞和組織不同水平進(jìn)行驗(yàn)證，鑒定該導(dǎo)向肽對(duì)中國(guó)人膀胱癌BIU-87細(xì)胞結(jié)合的特異性和親和性，為未來臨床上對(duì)膀胱腫瘤進(jìn)行早期診斷、術(shù)中準(zhǔn)確切除和開發(fā)靶向藥物提供定的理論基礎(chǔ)。 方法：將Matrigel基質(zhì)膠作為中國(guó)人膀胱癌BIU-87細(xì)胞的懸浮劑，用注射器將癌細(xì)胞種植于裸鼠腋窩背側(cè)皮下建立裸鼠移植瘤模型。通過化學(xué)方法合成小分子環(huán)七肽片段-SISSLTH，純化并標(biāo)記綠色熒光染料異硫氰酸熒光素FITC，利用免疫熒光技術(shù)原理，通過應(yīng)用流式細(xì)胞術(shù)和激光共聚焦顯微鏡技術(shù)兩種方法，對(duì)小分子多肽熒光探針FITC-SISSLTH在細(xì)胞和組織不同水平進(jìn)行驗(yàn)證，鑒定其與中國(guó)人膀胱癌細(xì)胞結(jié)合的特異性和親和性，以及該小分子探針在荷瘤裸鼠體內(nèi)的生物分布情況。 結(jié)果：在細(xì)胞水平對(duì)小分子多肽熒光探針FITC-SISSLTH進(jìn)行驗(yàn)證，其結(jié)果顯示：小分子多肽熒光探針FITC-SISSLTH與中國(guó)人膀胱癌BIU-87細(xì)胞系的親和性明顯高于人肝癌Hep-G2細(xì)胞系和人前列腺癌PC-3細(xì)胞系。利用激光共聚焦顯微鏡技術(shù)研究小分子探針在荷瘤裸鼠體內(nèi)的分布，實(shí)驗(yàn)結(jié)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(duì)膀胱癌組織有特異性和靶向性。 結(jié)論：在細(xì)胞水平對(duì)小分子多肽熒光探針FITC-SISSLTH進(jìn)行驗(yàn)證，其結(jié)果顯示：小分子多肽熒光探針FITC-SISSLTH與中國(guó)人膀胱癌BIU-87細(xì)胞系的親和性明顯高于人肝癌Hep-G2細(xì)胞系和人前列腺癌PC-3細(xì)胞系。利用激光共聚焦顯微鏡技術(shù)研究小分子探針在荷瘤裸鼠體內(nèi)的分布，實(shí)驗(yàn)結(jié)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(duì)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(xué)位授予單位】：山西醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R737.14

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