Cancer Molecular Pathology, School of Medicine, Griffith University, Gold Coast, QLD 4222, Australia.
Oncology, Gold Coast University Hospital, Gold Coast, QLD 4215, Australia.
Int J Mol Sci. 2020 Oct 20;21(20):7766. doi: 10.3390/ijms21207766.
The aim of the present study was to isolate and investigate the genetic heterogeneities in single circulating tumour cells (CTCs) from patients with colorectal carcinoma (CRC). Twenty-eight single CTCs were collected from eight patients with CRC using a negative immunomagnetic enrichment method. After validation with glyceraldehyde 3-phosphate dehydrogenase (GAPDH) gene expression in 3 colon cancer cell lines, a panel of 19 genes were used to analyse the single CTCs ( = 28), primary colorectal carcinoma tissues ( = 8) and colon carcinoma cells ( = 6) using real-time qPCR. Genetic heterogeneities were assessed by comparing gene expression profiles of single CTCs from the different patients and in the same patient, respectively. Genetic profiling of the single CTCs showed extensive heterogeneities of the selected genes among the CTCs. Hierarchical clustering analyses exhibited two clusters of CTCs with differentially expressed genes, which highlighted different modifications from the primary carcinomas. Further, the genetic heterogeneities were observed between different patients or in the same patient. Finally, expression was significantly ( = 0.0129) higher in single CTCs from CRC of advanced pathological stages (III or IV) CRC than in CTCs from CRC of early stages (I or II). Our findings suggest that single-cell genetic analysis can monitor the genetic heterogeneities and guide the personalised therapeutic targets in clinical sectors.
本研究旨在分离并研究来自结直肠癌(CRC)患者的单个循环肿瘤细胞(CTC)中的遗传异质性。使用负免疫磁珠富集法从 8 名 CRC 患者中收集了 28 个单个 CTC。在 3 种结肠癌细胞系中通过甘油醛 3-磷酸脱氢酶(GAPDH)基因表达验证后,使用实时 qPCR 分析了 19 个基因的panel(= 28),原发性结直肠癌组织(= 8)和结肠癌细胞(= 6)。通过比较不同患者和同一患者的单个 CTC 的基因表达谱来评估遗传异质性。单个 CTC 的遗传分析显示,所选基因在 CTC 之间存在广泛的异质性。层次聚类分析显示,具有差异表达基因的 CTC 分为两个簇,突出了与原发性癌不同的修饰。此外,在不同患者或同一患者之间观察到遗传异质性。最后,表达水平在晚期(III 或 IV)CRC 中明显(= 0.0129)高于早期(I 或 II)CRC 中的 CTC。我们的研究结果表明,单细胞遗传分析可以监测遗传异质性,并为临床领域的个体化治疗靶点提供指导。