Doulabi Ehsan Manouchehri, Dubois Louise, Löf Liza, Sinha Tanay Kumar, Harinck George Mickhael, Stålhandske Per, Larsson Anders, Kamali-Moghaddam Masood
Department of Immunology, Genetics & Pathology, Science for Life Laboratory, Uppsala University, SE-751 08, Uppsala, Sweden.
Department of Medical Sciences, Clinical Chemistry, Uppsala University, SE-751 85, Uppsala, Sweden.
Biochem Biophys Rep. 2024 Jun 21;39:101761. doi: 10.1016/j.bbrep.2024.101761. eCollection 2024 Sep.
Extracellular vesicles (EVs), whose main subtypes are exosomes, microparticles, and apoptotic bodies, are secreted by all cells and harbor biomolecules such as DNA, RNA, and proteins. They function as intercellular messengers and, depending on their cargo, may have multiple roles in cancer development. Thymidine kinase 1 (TK1) is a cell cycle-dependent enzyme used as a biomarker for cell proliferation. TK1 is usually elevated in cancer patients' serum, making the enzyme a valuable tumor proliferation biomarker that strongly correlates with cancer stage and metastatic capabilities. Here, we investigated the presence of TK1 in EVs derived from three prostate cancer cell lines with various p53 mutation statuses (LNCaP, PC3, and DU145), EVs from the normal prostate epithelial cell line RWPE-1 and EVs isolated from human seminal fluid (prostasomes). We measured the TK1 activity by a real-time assay for these EVs. We demonstrated that the TK1 enzyme activity is higher in EVs derived from the malignant cell lines, with the highest activity from cells deriving from the most aggressive cancer, compared to the prostasomes and RWPE-1 EVs. The measurement of TK1 activity in EVs may be essential in future prostate cancer studies.
细胞外囊泡（EVs）的主要亚型为外泌体、微颗粒和凋亡小体，由所有细胞分泌，含有DNA、RNA和蛋白质等生物分子。它们作为细胞间信使发挥作用，根据其携带的物质，可能在癌症发展中具有多种作用。胸苷激酶1（TK1）是一种细胞周期依赖性酶，用作细胞增殖的生物标志物。TK1通常在癌症患者血清中升高，使该酶成为一种有价值的肿瘤增殖生物标志物，与癌症分期和转移能力密切相关。在这里，我们研究了来自三种具有不同p53突变状态的前列腺癌细胞系（LNCaP、PC3和DU145）的EVs、正常前列腺上皮细胞系RWPE - 1的EVs以及从人精液中分离的EVs（前列腺小体）中TK1的存在情况。我们通过实时测定法测量了这些EVs的TK1活性。我们证明，与前列腺小体和RWPE - 1 EVs相比，来自恶性细胞系的EVs中TK1酶活性更高，其中来自最具侵袭性癌症的细胞的EVs活性最高。测量EVs中的TK1活性在未来前列腺癌研究中可能至关重要。
