Lapin Morten, Tjensvoll Kjersti, Oltedal Satu, Javle Milind, Smaaland Rune, Gilje Bjørnar, Nordgård Oddmund
Department of Haematology and Oncology, Stavanger University Hospital, N-4068, Stavanger, Norway.
Laboratory for Molecular Biology, Stavanger University Hospital, N-4068, Stavanger, Norway.
BMC Cancer. 2017 May 31;17(1):390. doi: 10.1186/s12885-017-3385-3.
Single-cell mRNA profiling of circulating tumour cells may contribute to a better understanding of the biology of these cells and their role in the metastatic process. In addition, such analyses may reveal new knowledge about the mechanisms underlying chemotherapy resistance and tumour progression in patients with cancer.
Single circulating tumour cells were isolated from patients with locally advanced or metastatic pancreatic cancer with immuno-magnetic depletion and immuno-fluorescence microscopy. mRNA expression was analysed with single-cell multiplex RT-qPCR. Hierarchical clustering and principal component analysis were performed to identify expression patterns.
Circulating tumour cells were detected in 33 of 56 (59%) examined blood samples. Single-cell mRNA profiling of intact isolated circulating tumour cells revealed both epithelial-like and mesenchymal-like subpopulations, which were distinct from leucocytes. The profiled circulating tumour cells also expressed elevated levels of stem cell markers, and the extracellular matrix protein, SPARC. The expression of SPARC might correspond to an epithelial-mesenchymal transition in pancreatic circulating tumour cells.
The analysis of single pancreatic circulating tumour cells identified distinct subpopulations and revealed elevated expression of transcripts relevant to the dissemination of circulating tumour cells to distant organ sites.
循环肿瘤细胞的单细胞mRNA分析可能有助于更好地理解这些细胞的生物学特性及其在转移过程中的作用。此外，此类分析可能揭示癌症患者化疗耐药性和肿瘤进展潜在机制的新知识。
通过免疫磁珠去除法和免疫荧光显微镜技术，从局部晚期或转移性胰腺癌患者中分离出单个循环肿瘤细胞。采用单细胞多重逆转录定量聚合酶链反应分析mRNA表达。进行层次聚类和主成分分析以识别表达模式。
在56份检测血样中的33份（59%）中检测到循环肿瘤细胞。完整分离的循环肿瘤细胞的单细胞mRNA分析揭示了上皮样和间充质样亚群，它们与白细胞不同。分析的循环肿瘤细胞还表达高水平的干细胞标志物和细胞外基质蛋白SPARC。SPARC的表达可能对应于胰腺循环肿瘤细胞中的上皮-间充质转化。
对单个胰腺循环肿瘤细胞的分析确定了不同的亚群，并揭示了与循环肿瘤细胞向远处器官部位扩散相关的转录本的高表达。
