基于细胞化学组分差异发展新型前列腺癌单细胞识别方法

Prostate cancer is the most common form of cancer in men and the second leading cause of cancer deaths, exceeded only by lung cancer. It is not satisfactory for the serum prostate-specific antigen (PSA) test both in sensitivity and specificity, which has been widely used for the early detection of prostate cancer currently.Based on the phenomenon that there is dramatic reduction in the zinc content of prostate tissue, and the expression differences of proteins with rich helix in the membrane of cells during the carcinogenic processes of prostate, this project will design and synthesis fluorescence probes with specific recognition of zinc ions, benzothiazole and tricarbocyanine derivatives, along with the multi-parametric fluorescence probes benzoxazoles, which are sensitive to proteins with rich helix domains, for the reason that they will undergo excited state intramolecular proton transfer (ESIPT) reaction and then produce new fluorescence emission signal when located in proteins with rich helix, furthermore, the intensity of the emission responds to the expression differences of proteins with rich helix. We will culture the two fluorescent probes based to different targets with normal prostate cells and prostate cancer cells together, and then develop new single-cell recognition methods for prostate cancer through double-probe mode and multicolor imaging.

前列腺癌是男性最常见的恶性肿瘤，死亡率紧随肺癌之后位居第二。目前被广泛应用于前列腺癌早期诊断的前列腺特异性抗原检测在敏感性和特异性两方面均不够理想。本项目基于前列腺细胞在癌变过程中锌离子浓度显著降低，以及细胞膜上富含螺旋的蛋白表达差异，设计合成特异性识别锌离子的荧光分子苯并噻唑类和三碳菁类衍生物，和对富含螺旋的蛋白质敏感的多参数荧光探针苯并噁唑类化合物，后一类化合物由于在富含螺旋的蛋白质环境中发生激发态分子内质子转移，产生新的荧光发射峰，而该发射峰的强度正好能反映富含螺旋的蛋白质表达的差异。我们将这两种基于不同靶标的荧光探针共同与前列腺正常细胞和癌细胞作用，通过双探针模式和多颜色成像技术发展新型前列腺癌单细胞识别的方法。

前列腺癌是男性生殖泌尿系统常见的恶性肿瘤之一，目前被广泛应用于前列腺癌早期诊断的前列腺特异性抗原检测在敏感性和特异性两方面均不够理想。本项目基于前列腺细胞在癌变过程中锌离子浓度显著降低，以及细胞膜上富含alpha-螺旋结构蛋白的表达差异，设计合成了特异性识别游离锌离子的3-羟基黄酮类荧光分子3HC-DPA，和对富含螺旋的蛋白质敏感的多参数荧光探针苯并噁唑类化合物1。后者由于激发态分子内质子转移（ESIPT）效应，通过识别细胞膜上富含alpha-螺旋结构的蛋白质，达到区分前列腺癌细胞的目的；而前者和锌离子结合后基于ESIPT屏蔽和光致电子转移（PET）机理，特异性识别细胞内游离态的锌离子，也可应用于前列腺癌细胞的识别。该部分研究成果发表在国际著名学术刊物Nucleic Acids Res.（IF：9.202）和Org. & Biomol. Chem.（IF：3.559）上。此外，我们还进一步在氧化应激和核酸损伤层面上探讨前列腺癌发生的诱因，初步筛选出了能够抑制前列腺癌细胞生长和发展的小分子化合物。结果显示，我们合成的SOD1抑制剂LD100可以增大前列腺癌细胞DU145处于G1期的比例，即有效阻滞癌细胞的生长，使其停留在细胞分裂的间期，而对正常前列腺细胞RWPE1影响不大。并且，相比于正常前列腺细胞，LD100对前列腺癌细胞DU145的凋亡更显著，因此LD100可作为一个有抗癌潜力的小分子进一步深入研究。