A biomarker for assisting the diagnosis of bone metastasis in lung adenocarcinoma

Abstract

The present invention discloses a biomarker for assisting in the diagnosis of bone metastasis of lung adenocarcinoma. The present invention specifically provides an application of a biomarker or a detection reagent thereof in the preparation of a product for assisting in the diagnosis or early screening of bone metastasis of lung adenocarcinoma, wherein the biomarker is AA and / or ANGPTL4. The present invention applies AA and / or ANGPTL4 to the diagnosis of bone metastasis of lung adenocarcinoma for the first time, and provides a simple, non-invasive, and long-term available detection method through serum testing; compared with the existing technology, the present invention can achieve earlier diagnosis of bone metastasis, and avoids the health risks brought by radiation, and has a high clinical application value; in short, the present invention provides a new biomarker combination for the early diagnosis of bone metastasis of lung adenocarcinoma, which is expected to provide more accurate guidance for the clinical treatment of patients, and is of great significance for the long-term monitoring of tumor metastasis.

Description

Technical Field

[0001] The present invention relates to a biomarker for auxiliary diagnosis of lung adenocarcinoma bone metastasis, belonging to the technical field of biomedical detection. Background Art

[0002] Lung adenocarcinoma is one of the leading causes of cancer death worldwide, and bone is a common site of metastasis. However, there is no simple and effective evaluation indicator for the active state of tumor cells in bone tissue. Existing imaging examinations often show positive results only after irreversible damage to the bone has occurred. At the same time, due to the radiation hazards and sensitivity of the methods, imaging examinations are difficult to use as early diagnosis of bone metastasis in patients and long-term dynamic monitoring of bone metastasis progression. Abnormal lipid metabolism is a key link in the proliferation and formation of metastases of tumor cells in target organs, and the application of biochemical molecules related to its mechanism in tumor metastasis is still lacking.

[0003] Therefore, the prior art still has the following shortcomings:

[0004] 1. Existing imaging examination methods, such as X-rays, are not sensitive enough to detect bone metastasis in a timely manner, leading to delayed treatment.

[0005] 2. Existing imaging examination methods, such as CT, ECT, PET-CT, etc., cause significant radiation damage to patients and are difficult to perform long-term continuous dynamic monitoring.

[0006] 3. Existing imaging examination methods require expensive dedicated machines, which affects the accessibility of examination methods in primary medical institutions (such as community health service centers).

[0007] 4. There is currently a lack of reliable serological biomarkers for auxiliary diagnosis and progression monitoring of bone metastasis of lung adenocarcinoma. Summary of the Invention

[0008] The present invention aims to provide a novel combination of serological markers for the diagnosis of bone metastasis of lung adenocarcinoma, and to solve the technical problem of the lack of early diagnostic markers and progression monitoring markers for bone metastasis of lung adenocarcinoma.

[0009] In order to achieve the above object, the present invention adopts the following technical solutions:

[0010] In a first aspect, the present invention provides a use of a biomarker or a detection reagent thereof in the preparation of a product for auxiliary diagnosis or early screening of lung adenocarcinoma bone metastasis, wherein the biomarker is AA and / or ANGPTL4.

[0011] Preferably, the detection reagent comprises a reagent for quantitatively detecting a biomarker.

[0012] Preferably, the detection reagent comprises a reagent for detecting a biomarker at the gene level and / or protein level.

[0013] Preferably, the detection reagent is a reagent used in one or more detection techniques or methods selected from the following group: enzyme-linked immunosorbent assay, immunofluorescence, radioimmunoassay, immunoprecipitation, immunoblotting, high performance liquid chromatography, capillary gel electrophoresis, near-infrared spectroscopy, mass spectrometry, immunochemiluminescence, colloidal gold immunoassay, fluorescence immunochromatography, surface plasmon resonance, immuno-PCR or biotin-avidin technology.

[0014] Preferably, the product comprises at least one of a reagent, a kit, a test paper and a chip.

[0015] In a second aspect, the present invention provides a biomarker combination comprising AA and ANGPTL4.

[0016] In a third aspect, the present invention provides a product comprising reagents for detecting the biomarker combination described in the second aspect of the present invention.

[0017] Preferably, the product is a kit, which further comprises a vector recording the following prediction model:

[0018] logit(P)=ln(P / (1-P))=-7.2234+0.3423AA+0.0030ANGPTL4;

[0019] Where P is the probability of bone metastasis in subjects with lung adenocarcinoma, AA and ANGPTL4 represent the concentrations of AA and ANGPTL4 in the serum samples of the subjects as detected by ELISA, respectively. The unit of AA concentration is pmol / mL, and the unit of ANGPTL4 concentration is pg / mL.

[0020] Preferably, the product is a diagnostic system or device, which includes a sample collection device, a sample detection device and a diagnostic device; wherein:

[0021] The sample collection device is configured as a device for collecting a blood sample from a subject, wherein the subject is a lung adenocarcinoma patient;

[0022] The sample detection device is a device capable of detecting the content or expression of AA and / or ANGPTL4 protein or nucleic acid thereof in the blood sample;

[0023] The diagnostic device includes a data acquisition module and a diagnostic module. The data acquisition module is configured to acquire data detected by the sample detection device. The diagnostic module is configured to determine whether bone metastasis occurs in the subject based on the data acquired by the data acquisition module.

[0024] Preferably, the diagnosis module is provided with the following diagnosis model:

[0025] logit(P)=ln(P / (1-P))=-7.2234+0.3423AA+0.0030ANGPTL4;

[0026] The data acquired by the data acquisition module is input into the diagnostic model to obtain the probability value of the subject having bone metastasis.

[0027] Compared with the prior art, the present invention has the following beneficial effects:

[0028] (1) The present invention proposes that the detection of serum arachidonic acid (AA) and angiogenesis-like protein 4 (ANGPTL4), alone or in combination, can assist in determining the active state of tumor cells in bone tissue. It is a novel marker for bone metastasis of lung adenocarcinoma and has good value in auxiliary diagnosis and progression monitoring.

[0029] (2) The present invention applies AA and / or ANGPTL4 to the diagnosis of bone metastasis of lung adenocarcinoma for the first time, and provides a simple, non-invasive, and long-term detection method through serum detection. Compared with the existing technology, the present invention can achieve earlier diagnosis of bone metastasis and avoid the health risks brought by radiation, and has a high clinical application value. In short, the present invention provides a new biomarker combination for the early diagnosis of bone metastasis of lung adenocarcinoma, which is expected to provide more accurate guidance for the clinical treatment of patients and is of great significance for the long-term monitoring of tumor metastasis. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 Serum AA and ANGPTL4 levels in healthy individuals and patients with different lung adenocarcinomas.

[0031] Figure 2 The ROC curve was used to analyze the diagnostic value of AA.

[0032] Figure 3 The ROC curve was used to analyze the diagnostic value of ANGPTL4.

[0033] Figure 4 The ROC curve was used to analyze the combined diagnostic value of AA and ANGPTL4. DETAILED DESCRIPTION

[0034] To make the present invention more clearly understood, preferred embodiments are described in detail below with reference to the accompanying drawings.

[0035] Based on the biological mechanism of AA and / or ANGPTL4 being related to lipid metabolism and bone metastasis, the present invention found that the concentrations of both in the serum of patients with lung adenocarcinoma bone metastasis were significantly increased and could be detected by ELISA method; the results of the examples of the present invention showed that the detection of AA and / or ANGPTL4 could effectively distinguish patients with lung adenocarcinoma bone metastasis from other types of metastasis or no metastasis, with an AUC value of 0.842 to 0.885, showing high diagnostic specificity and sensitivity; the present invention provides a new biomarker combination that can avoid the radiation damage caused by traditional imaging examinations, and is suitable for the early diagnosis of lung adenocarcinoma bone metastasis and long-term continuous and dynamic monitoring of patients' condition changes.

[0036] The ELISA method involves extracting patient serum and centrifuging it, then detecting the serum levels of AA and ANGPTL4 using ELISA. Concentrations are calculated based on a standard curve and a set cutoff value is used to determine whether the patient has bone metastasis. A value above this cutoff value is considered bone metastasis, while a value other than this cutoff value is considered non-bone metastasis (including no metastasis or other metastasis). The present invention measured the concentrations of AA and ANGPTL4 in different groups of lung adenocarcinoma patients and established a diagnostic model for lung adenocarcinoma bone metastasis using logistic regression analysis. The diagnostic performance of this model was validated using a receiver operating characteristic (ROC) curve.

[0037] Unless otherwise specified, the reagents, methods, and equipment used in the present invention are conventional in the art. Unless otherwise specified, the reagents and materials used in the following examples were commercially available. Experimental methods without specific conditions were generally carried out under conventional conditions or conditions recommended by the manufacturer.

[0038] Example

[0039] The study included healthy subjects, lung adenocarcinoma patients without metastasis, lung adenocarcinoma patients with other metastases but without bone metastasis, and lung adenocarcinoma patients with bone metastasis. Serum was collected and assayed for AA and ANGPTL4 levels using the following ELISA procedure. A total of 20 healthy subjects, 34 lung adenocarcinoma patients without metastasis, 31 lung adenocarcinoma patients with other metastases but without bone metastasis, and 71 lung adenocarcinoma patients with bone metastasis were included. ANGPTL4 was assayed in 71 subjects; AA was assayed in 58 of these subjects due to insufficient sample size.

[0040] ELISA method was used to detect the levels of AA and ANGPTL4 in serum:

[0041] 1. Sample Collection: Draw venous blood from the patient into a blood collection tube containing separation gel. Centrifuge at 3500 rpm for 15 minutes. Aspirate the upper serum layer and centrifuge at 13000 rpm for 10 minutes. Collect the supernatant. Store in a 4°C refrigerator for testing on the same day. Alternatively, store in a -80°C refrigerator for long-term storage. Centrifuge again at 13000 rpm for 10 minutes before use and collect the supernatant.

[0042] 2. ELISA Procedure: Coat AA and ANGPTL4 antibodies separately in a well plate, add patient serum to each well, incubate at 37°C for 1 hour, and wash the plate 3-5 times with deionized water for 3-5 minutes each. Add AA and ANGPTL4 enzyme-labeled antibodies, incubate at 37°C for 30 minutes, and wash the plate 3-5 times with deionized water for 3-5 minutes each. Add developer solution, incubate at 37°C for 15 minutes, add stop solution, and measure absorbance at 450 nm using a microplate reader. For each assay, use a standard to construct a standard curve.

[0043] 3. Analysis of test results: The concentrations of AA and ANGPTL4 in patient serum were calculated based on the standard curve.

[0044] The test results showed that the serum AA and ANGPTL4 concentrations in patients with lung adenocarcinoma bone metastasis were higher than those in patients with lung adenocarcinoma without metastasis, and in patients with lung adenocarcinoma with other metastases but without bone metastasis ( Figure 1 ).

[0045] A diagnostic model for lung adenocarcinoma bone metastasis was established to distinguish patients with lung adenocarcinoma without bone metastasis (including lung adenocarcinoma without metastasis and other metastases) from patients with lung adenocarcinoma bone metastasis. ROC curve analysis was used to evaluate the diagnostic performance of AA and ANGPTL4 as diagnostic markers, respectively, and in combination. It was found that the AUC for AA alone was 0.855 ( Figure 2 ), the optimal cutoff value was 15.0427, corresponding to a sensitivity of 96.55% and a specificity of 67.69%; the AUC of ANGPTL4 alone was 0.842 ( Figure 3 ), the optimal cutoff value was 393.989, corresponding to a sensitivity of 76.06% and a specificity of 72.97%. Logistic regression analysis was used to establish a diagnostic model for lung adenocarcinoma bone metastasis using AA and ANGPTL4. The prediction model equation was logit(P) = ln(P / (1-P)) = -7.2234 + 0.3423AA + 0.0030ANGPTL4. Receiver operating characteristic (ROC) analysis was performed based on the prediction results of the equation. The combined use of the two methods had the highest AUC of 0.885 ( Figure 4 ), with an optimal cutoff value of 0.4307872, corresponding to a sensitivity of 91.7% and a specificity of 88.9%. Therefore, AA and ANGPTL4 have high diagnostic value both individually and in combination, with their combined diagnostic value being even higher.

[0046] The above description is only a preferred embodiment of the present invention and does not constitute any formal or substantial limitation to the present invention. It should be noted that ordinary technicians in this technical field can make several improvements and supplements without departing from the present invention, and these improvements and supplements should also be regarded as the scope of protection of the present invention.

Claims

1. Use of a biomarker detection reagent in the preparation of a product for auxiliary diagnosis or early screening of lung adenocarcinoma bone metastasis, characterized in that: The biomarkers are arachidonic acid AA and / or angiogenesis-like protein 4 ANGPTL4; The detection reagent is a reagent for detecting biomarkers at the protein level.

2. The use according to claim 1, characterized in that The detection reagents include reagents for quantitatively detecting biomarkers.

3. The use according to claim 1, characterized in that The detection reagent is a reagent used in one or more detection techniques or methods selected from the following groups: enzyme-linked immunosorbent assay, immunofluorescence, radioimmunoassay, immunocoprecipitation, immunoblotting, high performance liquid chromatography, capillary gel electrophoresis, near-infrared spectroscopy, mass spectrometry, immunochemiluminescence, colloidal gold immunoassay, fluorescent immunochromatography, surface plasmon resonance, immuno-PCR or biotin-avidin technology.

4. The use according to any one of claims 1 to 3, characterized in that The product includes at least one of a reagent, a test kit, a test paper and a chip.

5. The use according to claim 4, characterized in that The product is a diagnostic system or device, which includes a sample collection device, a sample detection device, and a diagnostic device; wherein: The sample collection device is configured as a device for collecting a blood sample from a subject, wherein the subject is a lung adenocarcinoma patient; The sample detection device is a device capable of detecting the content of arachidonic acid AA and / or angiogenesis-like protein 4 ANGPTL4 protein in the blood sample; The diagnostic device includes a data acquisition module and a diagnostic module. The data acquisition module is configured to acquire data detected by the sample detection device. The diagnostic module is configured to determine whether bone metastasis occurs in the subject based on the data acquired by the data acquisition module.

6. The use according to claim 5, characterized in that The diagnostic module is provided with the following diagnostic models: logit(P)= ln(P / (1-P))=-7.2234+0.3423AA+0.0030ANGPTL4; The data acquired by the data acquisition module is input into the diagnostic model to obtain the probability value of the subject having bone metastasis.

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