精确测量细胞周期的微流控影像计数平台及其对胶质瘤干细胞分型的研究

Cell cycle regulation is closely related to cancer development. It is thus essential in cancer research to define the cell cycle phases at which the cancer cells reside. However, there has been no research on the precise cell cycle measurements of cancer stem cells. For a small number of cells, it is difficult to analyze the cell cycles by the traditional flow cytometry. To build an accurate cell cycle measurement platform with very small number of glioma stem cells (GSC), we will utilize our previously established microfluidic image cytometry (MIC) to measure the DNA contents in GSC, with simultaneous quantification of 4 cell cycle related proteins. This platform can precisely measure the cell cycles of every GSC and create GSC subtypes based on their cell cycles. Meanwhile, the influences of clonal passages, seeding intensities, and the expression of CD133 on the cell cycles of GSC will be studied. When associated with patient prognosis information and sphere formation assays, this platform will clarify the connections of GSC cell cycles to their malignancy. Therefore, analyzing the GSC subtypes by cell cycles is important to understand the tumorigenesis and development of GSC, clinical diagnosis of glioblastoma, and the subsequent therapies.

细胞周期调控机制与肿瘤的发生密切相关，因此了解肿瘤细胞所处的细胞周期对肿瘤研究至关重要。但是至今没有关于肿瘤干细胞细胞周期精确测量的研究。对于少量细胞，很难用传统的流式细胞仪进行细胞周期分析。因此，我们将之前建立的微流控影像计数技术用于细胞的DNA含量检测，拟同时结合4种细胞周期相关蛋白的定量检测，创建一个对少量胶质瘤干细胞进行精确细胞周期检测的平台。通过对每个胶质瘤干细胞所处细胞周期的精确检测，对胶质瘤干细胞进行基于细胞周期的精确分型，并研究克隆传代、接种密度、CD133表达对细胞周期的影响。结合病人预后信息和细胞球生成实验，可以更清晰地了解胶质瘤干细胞所处细胞周期与肿瘤恶性度的关系。因此对胶质瘤干细胞进行细胞周期亚型分析，对认识胶质瘤干细胞的发生和演化，胶质母细胞瘤的临床诊断与治疗具有十分重要的意义。

细胞周期调控机制与肿瘤的发生密切相关，因此了解肿瘤细胞所处的细胞周期对肿瘤研究至关重要。我们将之前建立的微流控影像计数技术用于细胞的DNA含量检测，创建了一个对少量细胞进行精确细胞周期检测的平台。我们研究发现，显微镜的放大倍数和细胞核染料的浓度对细胞周期的检测影响较大，显微镜的曝光时间和细胞个数对细胞周期检测的影响不大。我们还发现在蛋白染色的过程中，圆形细胞的蛋白荧光信号要优于铺展形细胞的蛋白信号。我们将胶质瘤干细胞按照4种蛋白表达进行细胞分型，发现细胞亚群与细胞球生成、病人预后、肿瘤恶性度都有密切的关系。对胶质瘤干细胞进行细胞亚型分析，对认识胶质瘤干细胞的发生和演化，胶质母细胞瘤的临床诊断与治疗具有十分重要的意义。项目资助发表SCI论文5篇，其中一区论文1篇，二区论文1篇，三区论文2篇，四区论文1篇，获批专利1项，待发表文章3篇，受理专利3项。培养研究生4名，2名已毕业，2名在读。培养本科生10余名，获得国家级和省级各类大学生创新创业大赛金奖银奖约7项。