纳米探针用于癌细胞整合素分子的原位标记和定量检测及探针的细胞生物学效应研究

The invasion and metastasis is the key step for prostate cancer progression and is the main reason to result the cancer patients death. Prognosis the invasion and metastasis tendency of early stage prostate cancer is urgently desired for prostate therapies now. The integrin molecules,α6β4 and α6β1, play important roles in prostate cancer invasion and metastasis at their earlier stage. In detail, the location, distribution, and protein express level of the two integrins work together to control the cancer cell invasion and metastasis. In this program,we will develop a series of nanoprobes which are composed of functional peptide and fluorescent noble metal cluster. The nanoprobe can target and quantify the integrins of prostate cancer cell of different pathological periods. Using the confocal microscopy, biological mass spectrometry,ICP-MS and combine with molecular biology technology to characterize physicochemical properties and biological effect of the nanoprobe, in situ mark and quantify the integrins in diferent pathological period cells..According the upper , we will declose the distribution and expression differences between the two integrins of prostate cancer cell in different pathological stage; And the relationship of the intrgrin abnormal expression and the invasion and/or metastasis ability of prostate cancer cell. This research results will provide the key data to judge invasion/matastasis tendency of the eralier stage prostate cancer, and to assist the clinician to make clinical surgery therapeutic schedule for patients.

侵袭、转移是早期前列腺癌恶化的重要生物学特征，是癌症患者死亡的重要原因。准确地判断早期前列腺癌细胞的侵袭、转移倾向，是前列腺癌治疗中急需解决的关键问题。前期研究提示：α6β4和α6β1整合素分子在前列腺细胞中分布及表达量，与早期肿瘤的侵袭和转移显著相关，但是如何实现对活细胞中整合素分子的原位标记和检测，是目前生物分析技术上挑战性的难题。本课题以前列腺癌活细胞中α6β4和α6β1为研究对象，发展一系列由多肽分子和金属团簇分子复合而成的纳米探针，利用活细胞成像技术和生物质谱技术，结合分子生物学手段，揭示探针的细胞生物学效应，实现对二种整合素在不同病理期癌细胞中的实时、原位标记和灵敏定量检测，建立活细胞内整合素表达量及分布情况与癌细胞的侵袭转移能力的依赖关系，为指导临床癌症分期、治疗方案的选择提供重要依据。

按照项目计划任务书，项目的研究内容为：发展一系列纳米探针，实现整合素在不同病理期癌细胞中的实时、原位标记和灵敏定量检测。项目完成了研究内容，主要工作为：成功制备了纳米探针并实现了细胞的原位标记，经过筛选确定了小肽序列Sv，并制备了铜团簇(Sv-Cu cluster)纳米探针，利用合成的Sv-Cu团簇对Hela细胞和A549细胞进行了单光子和双光子成像，证明该探针的双光子细胞成像应用价值。同时构建了肽分子-金团簇探针，发现探针可与细胞膜表面αIIbβ3整合素特异结合，利用激光烧蚀单细胞定量质谱，实现了多个单细胞结合的金团簇探针的定量，进一步换算为单细胞αIIbβ3整合素的绝对含量，通过数据分析表明，不同单细胞的αIIbβ3的含量有着较大的差异，实现了纳米探针对癌细胞上整合素的定量检测，整合素的表达量与癌症的发生和发展有密切关系，因此我们的方法为癌症的诊断与治疗提供了信息。