Urine protein markers for tumor bone metastasis and application thereof
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A bone metastasis and protein technology, applied in the field of clinical medicine, can solve problems such as unsatisfactory early diagnosis
Pending Publication Date: 2020-10-23
BEIJING NORMAL UNIVERSITY
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However, there are defects, and its early diagnosis is not ideal, and the disease is often present when it is discovered. See Sun Yan et al., Expert Consensus on the Diagnosis and Treatment of Bone Metastasis o
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Embodiment 1
[0033] Example 1. Establishment of recognized bone metastasis model
[0034] The Walker-256 cells used in this example were found in the mammary gland of the pregnant rat (Rattus norvegicus), which is considered a carcinosarcoma (see McEuen, C.S et al. (1933) Br. J. Exp. Pathol. 14, 384–391 and Simpkins, H et al (1991) Cancer Res. 51, 1334–1338).
[0035] Walker-256 is the most widely used transplantable tumor in experimental studies. The cells can cause obvious bone resorption and bone destruction at the position of rat bone implantation, which is consistent with the phenotype of bone metastasis in breast cancer patients, and is a bone metastasis model recognized in the art (see Shih, L.Y et al. (2004) J . Orthop. Res. 22, 1161–1167). In this example, the inventors planted walker-256 cells into the left tibia of rats to simulate tumor bone metastasis. Urine was collected before, at the beginning, and at the end of tumor growth for analysis.
[0036] 1. Materials and Reage...
Embodiment 2
[0047] Example 2. Sample collection and protein identification of urine
[0048] 1. Urine sample collection:
[0049] After modeling, rats in the experimental group and control group were placed in metabolic cages to collect urine on the 3rd, 5th, 7th, 10th, and 13th day. And 3 rats were taken, and CT was used to observe the rat tibial lesions.
[0050] 2. Extraction of protein in urine by ethanol precipitation
[0051] The method of ethanol precipitation of urinary protein refers to the method of Sun W et al. (see Sun W et al. Proteomics, 2005, 5(18):4994-5001). Urine centrifugation: 3000g / min, 4°C, 30min, take the supernatant; 12000g / min, 4°C, 30min. Take the supernatant, add 3-5 times pre-cooled ethanol, and precipitate overnight at 4°C. Centrifuge at 12000g / min at 4°C for 30min, and leave the precipitate to air dry at room temperature. Add about 500 μl of lysate to dissolve the precipitate by pipetting, and sonicate for 3 minutes. Centrifuge at 12000g / min at 4°C for ...
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Abstract
The invention relates to urine protein markers for tumor bone metastasis and application thereof. The invention relates to the application of a urine protein marker selected from a group consisting ofatrial natriuretic peptide receptor 2, galectin-3 binding protein, hepcidin, osteopontin, growth differentiation factor 15, hydroxy acid oxidase 2, NKG2-DII type integrated membrane protein, fatty acid binding protein, brain and complement component C6, alpha1 acidic glycoprotein, thioredoxin, mitochondria, beta-2-microglobulin, uridine and the like in the prediction of the risk of tumor bone metastasis and/or the diagnosis of tumor bone metastasis. According to the application, an animal model for screening the urine protein markers related to tumor bone metastasis is established, the urineprotein markers related to tumor bone metastasis are obtained by using the established animal model, and application of a detection reagent of the markers in preparation of a kit for diagnosing tumorbone metastasis of a subject is also established.
Description
technical field [0001] The present invention relates to the field of clinical medicine; specifically, the present invention relates to urine protein markers related to tumor bone metastasis. Specifically, the present invention relates to the use of a rat model of tumor bone metastasis and mass spectrometry analysis of urine protein markers to obtain urine protein markers related to human tumor bone metastasis, as well as the use of the urine protein markers. Background technique [0002] Bone is a common metastatic site of malignant tumors, only the lung and liver. Data show that 75% to 90% of patients with advanced tumors develop bone metastases and experience significant cancer pain, see Montiel-Ruiz RM et al. Bone cancer pain: from preclinical pharmacology to clinical trials. Gac Med Mex, 2013, 149(2) : 204-211. [0003] Tumor bone metastasis not only affects the quality of life of patients, but also causes patients to suffer from anemia, fractures, paraplegia, hypercal...
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