Otaş Hasan Ozan, Uluç Nasire, Demirkan İrem, Alkan Aylin, Yilmazer Açelya, Yaşa Seda, Khalafkhany Davod, Özören Nesrin, Ünlü Mehmet Burçin
Department of Molecular Biology and Genetics, Faculty of Arts and Sciences, Boğaziçi University, İstanbul, Turkey.
Department of Physics, Faculty of Arts and Sciences, Boğaziçi University, İstanbul, Turkey.
Turk J Biol. 2023 Jun 5;47(3):158-169. doi: 10.55730/1300-0152.2652. eCollection 2023.
BACKGROUND/AIM: Matrix metalloproteinases (MMPs) play an important role in the evaluation of many cancer types; however, the detection usually presents a challenge. Further assays for a better understanding of the fundamental roles of MMPs in pathophysiology are still needed. We aimed to use an activatable probe in scanning acoustic microscopy (SAM) to evaluate acoustically if the probe can aid the visualization of the effects of in vitro MMP activity.
We applied scanning acoustic impedance microscopy to obtain acoustic impedance maps of the cell line models of HT1080, THP-1, and SK-MEL-28 with and without MMPSense 680 probe incubation. We visually validated our results using confocal laser scanning microscopy imaging. We further analyzed the effects of MMPSense 680 probe on cell viabilities to eliminate any artifacts.
This is the first study presenting the applicability of SAM in the acoustical evaluation of MMPSense 680 probe cleavage in a cellular medium through acoustic impedance measurements. We proposed that SAM measurement with the activatable probe can be used as an effective tool for studying the acoustical variations of MMP activities in cell lines. As a result, we detected MMPSense 680 probe cleavage in HT1080 human fibrosarcoma cell line.
We showed that SAM with the smart probe can detect proteolytic activity using MMPSense 680 in in vitro HT1080 cell line by acoustic impedance measurements. SAM could be proposed as an alternative tool leading a novel way for a better understanding of the roles of MMPs in cancer progression before clinical settings.
背景/目的:基质金属蛋白酶(MMPs)在多种癌症类型的评估中发挥着重要作用;然而,其检测通常具有挑战性。仍需要进一步的检测方法以更好地理解MMPs在病理生理学中的基本作用。我们旨在使用扫描声学显微镜(SAM)中的可激活探针,通过声学方法评估该探针是否有助于体外MMP活性效应的可视化。
我们应用扫描声阻抗显微镜获取了HT1080、THP-1和SK-MEL-28细胞系模型在有无MMPSense 680探针孵育情况下的声阻抗图。我们使用共聚焦激光扫描显微镜成像对结果进行了视觉验证。我们进一步分析了MMPSense 680探针对细胞活力的影响,以消除任何假象。
这是第一项通过声阻抗测量展示SAM在细胞培养基中对MMPSense 680探针裂解进行声学评估的适用性的研究。我们提出,使用可激活探针的SAM测量可作为研究细胞系中MMP活性声学变化的有效工具。结果,我们在HT1080人纤维肉瘤细胞系中检测到了MMPSense 680探针的裂解。
我们表明,带有智能探针的SAM能够通过声阻抗测量在体外HT1080细胞系中利用MMPSense 680检测蛋白水解活性。在临床应用之前,SAM可被视为一种替代工具,为更好地理解MMPs在癌症进展中的作用开辟新途径。