Lung adenocarcinoma biomarker, detection system, detection kit and application

Abstract

The invention discloses a lung adenocarcinoma biomarker, a detection system, a detection kit and application, and belongs to the technical field of biology, and the biomarker is HROB. The lung adenocarcinoma marker detection system comprises a detection module and an analysis module, the detection module is used for detecting the content of a biomarker HROB in a sample; and the analysis module is used for receiving and analyzing the correlation between the data obtained by the detection module. The detection method is simple and low in cost.

Description

Technical Field

[0001] This invention relates to the field of biotechnology, and in particular to a lung adenocarcinoma biomarker, detection system, detection kit, and application. Background Technology

[0002] Lung adenocarcinoma is a major pathological type of lung cancer, and its incidence and mortality rates have been increasing year by year in recent years. Lung adenocarcinoma has an insidious onset, with few or no early symptoms, and most patients are diagnosed at an advanced stage, resulting in a poor prognosis. Currently, the diagnosis of lung adenocarcinoma mainly relies on imaging examinations (such as CT, PET-CT, etc.) and histopathological examinations, but these methods have certain limitations. Imaging examinations have limited accuracy in diagnosing early, small lesions, while histopathological examinations are invasive and may cause patients some pain and risks.

[0003] Biomarkers have significant applications in early disease diagnosis, disease monitoring, and prognostic assessment. Identifying sensitive and specific biomarkers for lung adenocarcinoma is crucial for early diagnosis, personalized treatment planning, and improved patient outcomes. While some biomarkers associated with lung adenocarcinoma have been discovered, such as carcinoembryonic antigen (CEA) and cytokeratin 19 fragment (CYFRA21-1), their sensitivity and specificity still need improvement. Therefore, the discovery of new biomarkers for lung adenocarcinoma is urgently needed. Summary of the Invention

[0004] One of the objectives of this invention is to provide a lung adenocarcinoma biomarker, detection system, detection kit, and application to address the problem that the lack of effective molecular indicators in the prior art for predicting metastasis risk and treatment response makes personalized clinical treatment difficult.

[0005] The present invention is achieved through the following technical solution: a lung adenocarcinoma biomarker, wherein the biomarker is HROB.

[0006] Furthermore, biomarkers are used for lung adenocarcinoma identification, disease monitoring, and prognostic assessment.

[0007] Furthermore, the expression levels of the biomarker were elevated in individuals with lung adenocarcinoma.

[0008] Another aspect of the present invention provides a detection system for lung adenocarcinoma biomarkers, the detection system comprising: a detection module and an analysis module; the detection module is used to detect the content of the biomarkers as described above in a sample; the analysis module is used to receive and analyze the correlation between the data obtained by the detection module.

[0009] Furthermore, the content detection module is used to detect HROB mRNA expression level by RT-qPCR and / or HROB protein level by immunohistochemistry or Western Blot.

[0010] Furthermore, the samples include, but are not limited to: tissue biopsy samples, liquid biopsy samples, and exfoliated cells from bronchoalveolar lavage fluid and / or sputum.

[0011] Furthermore, the analysis module is used to analyze the changes in the content of the biomarkers.

[0012] The present invention also provides a detection kit for lung adenocarcinoma, the kit comprising detection reagents, the detection reagents including reagents for detecting biomarkers as described above.

[0013] Finally, this invention provides an application of a lung adenocarcinoma biomarker, as described above.

[0014] Biomarkers or detection systems or kits include the following applications: (1) in the early diagnosis of lung adenocarcinoma; (2) in the prognostic assessment of lung adenocarcinoma; (3) in the targeted therapy targets of lung adenocarcinoma; (4) in the companion diagnostics of lung adenocarcinoma; (5) in the monitoring of treatment response in lung adenocarcinoma; (6) in the development of drugs for lung adenocarcinoma; (7) in the study of the pathogenesis of lung adenocarcinoma; and (8) in the preparation of products for the prognostic assessment of lung adenocarcinoma.

[0015] Compared with the prior art, the present invention has the following advantages and beneficial effects:

[0016] 1. The biomarkers of the present invention enrich the diversity of diagnostic biomarkers for lung adenocarcinoma. The AUC of the HROB biomarker of the present invention is 0.968, which is significantly correlated with the progression of lung adenocarcinoma and has high prognostic accuracy.

[0017] 2. The HROB biomarker provided by this invention greatly simplifies the detection method and has low cost. The kit containing the HROB biomarker provided by this invention can be used simultaneously for diagnosis (differentiating lung adenocarcinoma from benign lesions) and prognostic assessment, achieving the effect of "one test, two effects". Attached Figure Description

[0018] The accompanying drawings, which are included to provide a further understanding of embodiments of the invention and form part of this application, do not constitute a limitation thereof. In the drawings:

[0019] Figure 1 The diagram shows the analysis results provided in Embodiment 1 of the present invention.

[0020] Figure 2 This is a graph showing the high expression of HROB in lung adenocarcinoma as provided in Embodiment 1 of the present invention.

[0021] Figure 3 This is a graph showing the mRNA expression level of HROB provided in Example 1 of the present invention.

[0022] Figure 4 This is a graph showing the proliferation rate of the HROB silent group provided in Embodiment 1 of the present invention.

[0023] Figure 5 This is a micrograph of normal lung tissue provided in Embodiment 1 of the present invention.

[0024] Figure 6 This is a micrograph of lung cancer tissue provided in Embodiment 1 of the present invention.

[0025] Figure 7 This is a quantitative statistical analysis chart of immunohistochemical slide AB provided in Example 1 of the present invention. Detailed Implementation

[0026] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are some embodiments of the present invention, but not all embodiments.

[0027] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. While only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein may be used in the implementation or testing of this invention. All references to this specification are incorporated by way of citation to disclose and describe methods and / or materials associated with those references. In the event of any conflict with any incorporated reference, the content of this specification shall prevail. The terms “comprising,” “including,” “having,” “containing,” etc., as used herein are open-ended terms meaning to include but not limited to. Unless the context clearly indicates otherwise, the expressions “a” and “an” as used herein include plural references. The term “about” as used herein indicates a range of ±20% of the following numerical value. In some embodiments, the term “about” indicates a range of ±10% of the following numerical value. In some embodiments, the term “about” indicates a range of ±5% of the following numerical value.

[0028] Example 1

[0029] This embodiment discloses a biomarker for lung adenocarcinoma, addressing the limitations of existing biomarkers (such as CEA and CYFRA21-1) in terms of sensitivity / specificity, leading to insufficient early diagnosis of lung adenocarcinoma often occurring at an intermediate or advanced stage by imaging diagnosis. Furthermore, the lack of effective molecular indicators for predicting metastasis risk and treatment response hinders personalized clinical treatment and contributes to deficiencies in existing prognostic assessment systems. Moreover, known driver genes (EGFR / ALK, etc.) only cover a portion of the population, highlighting the urgent need for new therapeutic targets.

[0030] To address the aforementioned issues, this embodiment provides a novel use of the HROB gene or its encoded protein as a diagnostic / prognostic biomarker for lung adenocarcinoma.

[0031] First, bioinformatics analysis was performed on the HROB gene or its encoded protein. Gene expression profile data from lung adenocarcinoma tissue and normal lung tissue were collected, and differential analysis was conducted using bioinformatics software and algorithms. The results are as follows: Figure 1 As shown, in Figure 1 (a) is the LUAD-DEGs volcano plot of HROB, (b) is the pan-cancer expression of HROB in the TCGA database, (c) is the expression of HROB in TCGA-LUAD paired samples, and (d) is the pan-cancer expression of HROB in the UCSC XENA database.

[0032] Figure 2 This study demonstrates the high expression of HROB in lung adenocarcinoma. Figure 2 (a) shows the expression of HROB in GSE116959, (b) shows the expression of HROB in GSE43458, and (c) shows the expression of HROB in GSE18842.

[0033] By culturing lung adenocarcinoma cell lines and normal lung epithelial cell lines. Related experimental results are as follows: Figure 3 and Figure 4 As shown, from Figure 3 It can be seen that the mRNA expression level of HROB in lung adenocarcinoma cell lines (A549, NCI-H1299) is significantly higher than that in normal lung epithelial cell line (BEAS-2B). Figure 4 As can be seen, in the NCl-H1299 cell line, the proliferation rate of the HROB-silenced group (siRNA) was significantly lower than that of the unsilenced group (NC).

[0034] Lung adenocarcinoma tissue and adjacent normal tissue samples were collected, and the protein expression level of HROB in the tissues was detected by IHC method. Figure 5 A micrograph of normal lung tissue is shown. Figure 6 A micrograph of lung cancer tissue is shown. Figure 7 The quantitative statistical analysis graph of immunohistochemical sections AB is shown. From Figure 5 , 6 As can be seen from Figures 7 and 8, the positive expression rate of HROB in lung adenocarcinoma tissue is significantly higher than that in adjacent normal tissue.

[0035] As can be seen from the above, the biomarkers disclosed in this embodiment can effectively detect lung adenocarcinoma by collecting biological samples (such as exfoliated cells from pleural fluid, sputum, etc.) from suspected lung adenocarcinoma patients. Appropriate detection methods are used to detect the expression level of HROB in the samples. The detection results are compared with the expression level of normal control samples. If the expression level of HROB in the sample is significantly higher than that in the normal control samples, it suggests the possibility of lung adenocarcinoma.

[0036] Alternatively, prognostic assessment can be achieved using biomarkers as described in this embodiment. This involves collecting biological samples (such as tissue, exfoliated cells from pleural fluid, etc.) from patients with lung adenocarcinoma. The expression level of HROB in the samples is detected, and patients are divided into high-expression and low-expression groups based on the HROB expression level. Patients in the high-expression group have a poorer prognosis, and clinicians may consider more aggressive treatment plans; patients in the low-expression group have a better prognosis, and treatment plans can be adjusted appropriately.

[0037] Example 2

[0038] This embodiment discloses a specific method for screening and detecting samples using the lung adenocarcinoma biomarker described in Example 1. In this embodiment, HROB is used as the core biomarker, while β-actin is used as an internal reference gene (Housekeeping Gene) for standardization to ensure the reliability and comparability of detection results between different samples. Specifically, the primer sequences used in this embodiment are shown in Table 1.

[0039] Table 1. Primer sequence listing

[0040]

[0041] This embodiment discloses a novel use of the HROB gene or its encoded protein as a diagnostic / prognostic biomarker for lung adenocarcinoma.

[0042] The sample types in this embodiment include, but are not limited to, tissue biopsy samples (surgical / puncture samples), liquid biopsy samples (peripheral blood ctDNA, exosomes, circulating tumor cells), and bronchoalveolar lavage fluid / sputum exfoliated cells.

[0043] The detection kit in this embodiment contains detection components that specifically recognize HROB: a set of qPCR primers / probes for HROB mRNA; a monoclonal antibody against HROB protein; and a matching control.

[0044] The specific detection methods are as follows: at the nucleic acid level, RT-qPCR is used to detect HROB mRNA expression levels; at the protein level, immunohistochemistry (IHC) or Western blotting is used to detect HROB protein levels.

[0045] Screening of patient subgroups suitable for HROB-targeted therapy (such as siRNA / monoclonal antibody drugs) and monitoring of treatment response.

[0046] The specific embodiments described above further illustrate the purpose, technical solution, and beneficial effects of the present invention. It should be understood that the above description is only a specific embodiment of the present invention and is not intended to limit the scope of protection of the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. A biomarker for lung adenocarcinoma, characterized in that, The biomarker is HROB.

2. The biomarker according to claim 1, characterized in that, The biomarkers are used for lung adenocarcinoma identification, disease monitoring, and prognostic assessment.

3. The biomarker according to claim 1 or 2, characterized in that, The expression levels of the biomarker were elevated in individuals with lung adenocarcinoma.

4. A detection system for lung adenocarcinoma markers, characterized in that, The detection system includes: Detection module and analysis module; The detection module is used to detect the content of the biomarker described in claim 1 in the sample; The analysis module is used to receive and analyze the correlation between the data obtained by the detection module.

5. The detection system according to claim 4, characterized in that, The content detection module is used to detect HROB mRNA expression level and / or, using RT-qPCR. HROB protein levels were detected by immunohistochemistry or Western blotting.

6. The detection system according to claim 4, characterized in that, The samples include, but are not limited to: Tissue biopsy samples, liquid biopsy samples, and exfoliated cells from bronchoalveolar lavage fluid and / or sputum.

7. The detection system according to claim 4, characterized in that, The analysis module is used to analyze the changes in the content of the biomarkers.

8. A detection kit for lung adenocarcinoma, characterized in that, The kit includes detection reagents, which include reagents for detecting the biomarker of claim 1.

9. The use of the biomarker according to any one of claims 1 to 3, the detection system according to any one of claims 4 to 7, or the kit according to claim 8 in any one of the following: (1) Application in the early diagnosis of lung adenocarcinoma; (2) Application in prognostic assessment of lung adenocarcinoma; (3) Application in targeted therapy for lung adenocarcinoma; (4) Application in the companion diagnosis of lung adenocarcinoma; (5) Application in monitoring treatment response in lung adenocarcinoma; (6) Application in the development of drugs for lung adenocarcinoma; (7) Application in the study of the pathogenesis of lung adenocarcinoma; (8) Application in the preparation of products for evaluating the prognostic effects of lung adenocarcinoma.

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