Biomarker for auxiliary diagnosis of lung adenocarcinoma bone metastasis

Abstract

The invention discloses a biomarker for auxiliary diagnosis of lung adenocarcinoma bone metastasis. The invention specifically provides application of a biomarker or a detection reagent thereof in preparation of a product for auxiliary diagnosis or early screening of lung adenocarcinoma bone metastasis. The biomarker is AA and / or ANGPTL4. According to the invention, AA and / or ANGPTL4 are / is applied to diagnosis of lung adenocarcinoma bone metastasis for the first time, and a simple, non-invasive and long-term available detection method is provided through serum detection; compared with the prior art, earlier bone metastasis diagnosis can be achieved, health risks caused by radiation are avoided, and high clinical application value is achieved; in a word, the invention provides a new biomarker combination for early diagnosis of lung adenocarcinoma bone metastasis, is expected to provide more accurate guidance for clinical treatment of patients, and has important significance for long-term monitoring of tumor metastasis.

Description

Technical Field

[0001] The 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. At the same time, due to radiation hazards and method sensitivity, 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 metastasis of tumor cells in metastatic 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 disadvantages:

[0004] 1. Existing imaging examination methods, such as X-rays, are not sensitive enough to detect bone metastasis in time, 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, etc.).

[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 lung adenocarcinoma bone metastasis, and to solve the technical problem of the lack of early diagnosis markers and progression monitoring markers for lung adenocarcinoma bone metastasis.

[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, co-immunoprecipitation, 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.

[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] Wherein, P is the probability of bone metastasis in subjects with lung adenocarcinoma, AA and ANGPTL4 represent the concentrations of AA and ANGPTL4 in serum samples of subjects detected by ELISA method, 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 comprises a data acquisition module and a diagnostic module. The data acquisition module is configured to acquire data detected by the sample detection device, and the diagnostic module is configured to determine whether bone metastasis occurs in the subject according to 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 the determination of the active state of tumor cells in bone tissue, and is a new marker for bone metastasis of lung adenocarcinoma, with good auxiliary diagnosis and progression monitoring value;

[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 available 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 caused 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 people 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] In order 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 finds that the concentrations of the two in the serum of patients with lung adenocarcinoma bone metastasis are significantly increased and can be detected by the ELISA method; the results of the embodiments of the present invention show that the detection of AA and / or ANGPTL4 can effectively distinguish patients with lung adenocarcinoma bone metastasis from other types of metastasis or no metastasis, and the AUC value is 0.842-0.885, which has high diagnostic specificity and sensitivity; the present invention provides a new biomarker combination, which 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] Among them, the steps of ELISA method detection include: extracting patient serum and centrifuging it, using ELISA method to detect the levels of AA and ANGPTL4 in the serum respectively, and finally calculating the concentration according to the standard curve and determining whether the patient has bone metastasis according to the set cutoff value. If it is higher than this value, it is diagnosed as bone metastasis, otherwise it is diagnosed as no bone metastasis (including no metastasis or other metastasis). The embodiment of the present invention measured the concentration of AA and ANGPTL4 in different lung adenocarcinoma patient groups, and established a diagnostic model for lung adenocarcinoma bone metastasis by using logistic regression analysis, and verified the diagnostic performance of the model by ROC curve.

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

[0038] Example

[0039] Healthy physical examination subjects, patients with lung adenocarcinoma without metastasis, patients with lung adenocarcinoma with other metastases but without bone metastasis, and patients with lung adenocarcinoma with bone metastasis were selected from the Lingang Campus of the Sixth People's Hospital Affiliated to Shanghai Jiao Tong University School of Medicine. Serum was collected from patients, and the levels of AA and ANGPTL4 in serum were detected according to the following ELISA method steps. A total of 20 healthy physical examination subjects, 34 patients with lung adenocarcinoma without metastasis, 31 patients with lung adenocarcinoma with other metastases but without bone metastasis, and 71 patients with lung adenocarcinoma with bone metastasis were included (ANGPTL4 was tested in 71 patients, and AA was tested in 58 of them 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 3500rpm for 15 minutes, draw the upper serum and centrifuge at 13000rpm for 10 minutes, collect the supernatant, and store it in a 4-degree refrigerator for testing on the same day. Or store it in a -80-degree refrigerator for a long time, and centrifuge it again at 13000rpm for 10 minutes before use to collect the supernatant.

[0042] 2.ELISA operation steps: Coat AA and ANGPTL4 antibodies in the well plate, add patient serum, incubate at 37 degrees for 1 hour, and wash the plate with deionized water 3-5 times, 3-5 minutes each time. Add AA and ANGPTL4 enzyme-labeled antibodies, incubate at 37 degrees for 30 minutes, and wash the plate with deionized water 3-5 times, 3-5 minutes each time. Add developer, incubate at 37 degrees for 15 minutes, add stop solution, and use an enzyme reader to detect absorbance at 450nm. For each test, use the standard to draw a standard curve.

[0043] 3. Analysis of test results: The concentrations of AA and ANGPTL4 in the patient's 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 patients with lung adenocarcinoma with other metastases but without bone metastasis ( Figure 1 ).

[0045] A diagnosis model for lung adenocarcinoma bone metastasis was established to distinguish patients with lung adenocarcinoma without bone metastasis (including lung adenocarcinoma without metastasis and other metastasis) and patients with lung adenocarcinoma bone metastasis. ROC curve analysis was used to evaluate the diagnostic performance of AA and ANGPTL4 as diagnostic markers and in combination. It was found that the AUC of 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 AA and ANGPTL4 combined with lung adenocarcinoma bone metastasis. The prediction model equation was logit(P)=ln(P / (1-P))=-7.2234+0.3423AA+0.0030ANGPTL4. ROC analysis was performed based on the prediction results of the equation. The AUC was the highest when the two were used together, which was 0.885( Figure 4 ), the optimal cutoff value was 0.4307872, corresponding to a sensitivity of 91.7% and a specificity of 88.9%. Therefore, AA and ANGPTL4 have high diagnostic value when diagnosed alone or in combination, and the combined diagnostic value is even higher.

[0046] The above description is only a preferred embodiment of the present invention and is not any formal or substantial limitation of the present invention. It should be pointed out that a person skilled in the art can make several improvements and supplements without departing from the present invention, and these improvements and supplements should also be regarded as the protection scope of the present invention.

Claims

1. 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, characterized in that: The biomarkers are AA and / or ANGPTL4.

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 2, characterized in that: The detection reagents include reagents for detecting biomarkers at the gene level and / or protein level.

4. The use according to claim 3, 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.

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

6. A biomarker combination, characterized in that: The biomarker panel includes AA and ANGPTL4.

7. A product comprising reagents for detecting the biomarker combination of claim 6.

8. The use according to claim 5 or the product according to claim 7, characterized in that: The product is a kit, which also includes a carrier recording the following prediction model: logit(P)=ln(P / (1-P))=-7.2234+0.3423AA+0.0030ANGPTL4; Wherein, P is the probability of bone metastasis in subjects with lung adenocarcinoma, AA and ANGPTL4 represent the concentrations of AA and ANGPTL4 in serum samples of subjects detected by ELISA method, respectively. The unit of AA concentration is pmol / mL, and the unit of ANGPTL4 concentration is pg / mL.

9. The use according to claim 5 or the product according to claim 7, 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 or expression of AA and / or ANGPTL4 protein or nucleic acid thereof in the blood sample; The diagnostic device comprises a data acquisition module and a diagnostic module. The data acquisition module is configured to acquire data detected by the sample detection device, and the diagnostic module is configured to determine whether bone metastasis occurs in the subject according to the data acquired by the data acquisition module.

10. The use or product according to claim 9, characterized in that: The diagnosis module is provided with the following diagnosis model: 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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