研究血漿Ki-67、犬尿氨酸-3-單氧化脢和纖維蛋白原- 纖維蛋白降解產物作為犬癌症診斷的生物標誌物

Abstract

癌症生物標誌物被視為腫瘤診斷和預後預測的強而有力工具。這些分子由腫瘤和腫瘤微環境產生，因此存在於患者的循環系統中。腫瘤在形成過程中會引發因為組織破壞而導致止血反應的活化，進而促進腫瘤的生長和擴散。止血功能障礙常常在人類癌症患者中被檢測到，通常會導致血漿纖維蛋白原- 纖維蛋白分解產物（DR-70）水平升高。酶犬尿氨酸3-單加氧酶（KMO）在色氨酸代謝中扮演重要之作用，先前KMO已被證實為神經退行性疾病和惡性腫瘤的主要標記物。Ki-67核蛋白在細胞增殖期間增加，為腫瘤細胞進展之指標。本研究旨在評估犬癌症中DR-70、KMO 和Ki-67水平的應用於犬癌診斷之可行性。本研究共蒐集641個臨床腫瘤犬樣本，並以58個健康個體作為對照組。通過酶聯免疫吸附法（ELISA）測定了血漿中KMO、DR-70和Ki-67值的濃度。結果顯示，腫瘤犬中DR-70、KMO 和Ki-67的濃度明顯高於健康犬 (P<0.001)。 每個生物標誌物的ROC (AUC) 面積分別是DR-70為0.898 (P<0.001)， KMO 為0.809 (P<0.001) ，Ki-67為0.533 (P<0.05)。 與單一標誌物相比，三聯合檢測的AUC 值最高，初步判斷能有效增加腫瘤的診斷率(AUC 為0.934)。 在這三種生物標誌物中，我們發現不同的腫瘤類型，其表現量皆顯著高於健康對照組，包括淋巴瘤、乳腺腫瘤、黑色素瘤等。此外，在同一病患在不同時間檢測DR-70腫瘤標誌物的案例中，我們發現DR-70 的表現量與腫瘤進展相關，這表明其可作為追踪腫瘤發展具潛力的腫瘤標誌物。總結來說，這項研究建議DR-70、KMO 和Ki-67可作為犬癌症診斷和預測腫瘤發生的生物標記物，並且預期它們在未來臨床上的應用。這是目前唯一評估血液DR-70、KMO 和Ki-67在獸醫腫瘤學中作為診斷與癒後追蹤分子之研究。

Cancer biomarkers are viewed as powerful tools for tumor diagnosis and prognosis prediction. These molecules are produced by tumors and the tumor microenvironments for them could be detected in the patient’s circulation. Tumor cells elicit a chronic hemostatic activation, and the pro-coagulant activities facilitate tumor growth and dissemination. Hemostatic dysfunctions are commonly detected in human cancer patients and usually result in a high plasma fibrinogen-fibrin degradation product (DR-70) level. The enzyme kynurenine 3-monooxygenase (KMO), which plays a central role in tryptophan metabolism, has previously been identified as the main factor in neurodegenerative diseases and malignant tumors. Ki-67 protein increased during cell proliferation. Therefore, this study aims to evaluate the diagnostic application of DR-70, KMO, and Ki-67 levels in canine cancers. A total of 641 clinically neoplastic canine samples and 58 healthy individuals were enrolled. The levels of plasma KMO, DR-70, and Ki-67 values were determined by the enzyme-linked immunosorbent assay (ELISA). The results showed that the expressions of DR-70, KMO, and Ki-67 were significantly increased in tumor dogs than those in healthy dogs (P<0.001). The determination of the area under the ROC (AUC) for each biomarker is 0.898 for DR-70 (P<0.001), 0.809 for KMO (P<0.001), and 0.533 for Ki-67 (P<0.05). The AUC value of combined detection increases the diagnostic rate compared to the single marker (AUC, 0.934). Different tumor types exhibited various levels of these three biomarkers, including lymphoma, mammary gland tumor, melanoma, etc. Furthermore, DR-70 values were correlated to tumor progression in several cases, which indicates its potential for tracing tumor development. In summary, this study suggested that DR-70, KMO, and Ki-67 levels are possible biomarkers for canine cancer diagnosis and prognosis, and their clinical application is expected which is the first study to evaluate the clinical significance of DR-70, KMO, and Ki-67 expressions in veterinary oncology.