Borisov Alexey V, Zakharova Olga A, Samarinova Alisa A, Yunusova Natalia V, Cheremisina Olga V, Kistenev Yury V
Laboratory of Laser Molecular Imaging and Machine Learning, National Research Tomsk State University, 634050 Tomsk, Russia.
Laboratory of Tumor Biochemistry, Cancer Research Institute, Tomsk National Research Medical Center RAS, 634009 Tomsk, Russia.
Diagnostics (Basel). 2022 Jul 24;12(8):1792. doi: 10.3390/diagnostics12081792.
This study was aimed to investigate the applicability of the exosome fluorescence-lifetime imaging microscopy (FLIM) for colorectal cancer (CRC) diagnosis. Differential ultra-centrifugation was used to extract exosomes from the blood plasma of 11 patients with colon polyps (CPs) and 13 patients with CRC at the T2-4, N0-3, and M0-1 stages. Analysis was performed using a two-photon FLIM device. In total, 165 and 195 FLIM images were recorded for the CP and CCR patient groups, respectively. Two classes of exosomes differentiated by autofluorescence average lifetime tm were discovered in the samples. The first class of exosomes with tm = (0.21 ± 0.06) ns was mostly found in samples from CRC patients. The second class with tm = (0.43 ± 0.19) ns was mostly found in samples from CP patients. The relative number of “CRC-associated” exosomes Nch in the FLIM dataset was shown to be very small for the CP patient group and large for the CRC patient group. This difference was statistically significant. Therefore, the suggested CRS diagnostics criterion can be as follows. If Nch > 0.5, the probability of CRC is high. If Nch < 0.3, the probability of CRC is low.
本研究旨在探讨外泌体荧光寿命成像显微镜(FLIM)在结直肠癌(CRC)诊断中的适用性。采用差速超速离心法从11例结肠息肉(CP)患者和13例处于T2 - 4、N0 - 3和M0 - 1期的CRC患者的血浆中提取外泌体。使用双光子FLIM设备进行分析。分别为CP患者组和CRC患者组记录了165张和195张FLIM图像。在样本中发现了两类通过自发荧光平均寿命tm区分的外泌体。第一类外泌体tm =(0.21±0.06)ns,主要存在于CRC患者的样本中。第二类外泌体tm =(0.43±0.19)ns,主要存在于CP患者的样本中。FLIM数据集中“CRC相关”外泌体Nch的相对数量在CP患者组中显示非常少,而在CRC患者组中则很大。这种差异具有统计学意义。因此,建议的CRC诊断标准如下。如果Nch>0.5,则CRC的可能性高。如果Nch<0.3,则CRC的可能性低。