Korea Basic Science Institute, Seoul 02841, South Korea.
School of Chemical Engineering, Chonnam National University, Gwangju 61186, South Korea.
Bioconjug Chem. 2020 Mar 18;31(3):803-812. doi: 10.1021/acs.bioconjchem.9b00870. Epub 2020 Feb 24.
Lung cancer is diagnosed at an advanced stage due to its unrecognized symptoms, resulting in high mortality. In recent decades, research into the development of an early diagnostic method for lung cancer has expanded in order to overcome the high mortality rate. Calpain 2 (CAPN2) has been suggested as a tumor marker linked to angiogenesis, cell proliferation, and migration in non-small cell lung cancer. In this study, CAPN2 enzyme-activatable near-infrared peptide sensor linked to human serum albumin (HSA-CAPN2) was developed. Intracellular localization and strong recovered fluorescence signals of HSA-CAPN2 were observed in experiments using A549-Luc cells, and signal recovery was inhibited by ALLN (a CAPN2 inhibitor). distribution and signal recovery evaluations performed using A549-Luc cell xenograft mice revealed that HSA-CAPN2 accumulated in the tumor region and produced high fluorescent signal recovery. Three-dimensional reconstructed images using single-plane illumination microscopy after tissue clarity visualized localization of HSA-CAPN2 in tumors. In addition, ALLN pretreatment showed a significant inhibitory effect on signal recovery of HSA-CAPN2, and that inhibition was induced by downregulation of CAPN2 at the gene and protein levels followed by decreases in Ca levels. Overall, the results demonstrate the potential of HSA-CAPN2 as a sensor for CAPN2-enriched cancer.
肺癌由于其症状不被识别而在晚期被诊断出，导致死亡率很高。近几十年来，为了克服高死亡率，人们对开发肺癌早期诊断方法的研究不断扩大。钙蛋白酶 2（CAPN2）已被提出作为与非小细胞肺癌中的血管生成、细胞增殖和迁移相关的肿瘤标志物。在这项研究中，开发了与人血清白蛋白（HSA-CAPN2）连接的钙蛋白酶 2 酶激活近红外肽传感器。在使用 A549-Luc 细胞的实验中观察到 HSA-CAPN2 的细胞内定位和强恢复荧光信号，并且 ALLN（CAPN2 抑制剂）抑制信号恢复。在 A549-Luc 细胞异种移植小鼠中进行的分布和信号恢复评估表明，HSA-CAPN2 积聚在肿瘤区域并产生高荧光信号恢复。使用单平面照明显微镜进行三维重建图像后，组织清晰度可视化了 HSA-CAPN2 在肿瘤中的定位。此外，ALLN 预处理对 HSA-CAPN2 的信号恢复显示出显著的抑制作用，这种抑制是通过 CAPN2 在基因和蛋白质水平下调以及 Ca 水平降低诱导的。总的来说，这些结果表明 HSA-CAPN2 作为富含 CAPN2 的癌症传感器的潜力。
