Application of CXCL1, CXCL5 and CXCL11 in the preparation of a kit for diagnosing metastasis of esophageal squamous cell carcinoma

Abstract

The present invention relates to the field of medical detection. Specifically, the present invention provides a biomarker for esophageal squamous cell carcinoma metastasis and its application, wherein the biomarker includes three secretory proteins, namely CXCL1, CXCL5 and CXCL11. The above-mentioned markers are used to perform typing diagnosis of esophageal squamous cell carcinoma. When CXCL1, CXCL5 and CXCL11 are combined, the diagnostic power AUC is as high as 0.997. This provides mutual verification for pathological diagnosis results, ensures that the case diagnosis results are correct, and facilitates subsequent precise treatment.

Description

Technical Field

[0001] The present invention relates to the field of medical diagnosis, and in particular, to an application of CXCL1, CXCL5 and CXCL11 in preparing a kit for diagnosing metastasis of esophageal squamous cell carcinoma. Background Art

[0002] Esophageal cancer is a malignant tumor that seriously endangers people's health worldwide. ESCC is very prone to lymphatic metastasis. At present, the five-year clinical survival rate of esophageal cancer is extremely low, which is mainly closely related to ESCC metastasis. Therefore, the discovery of markers for diagnosing ESCC metastasis is the key to improving the overall efficacy of ESCC patients.

[0003] CXCL1 (Growth-regulated α protein, GROα) is an important member of the chemokine family and is expressed in macrophages, neutrophils and epithelial cells. CXCL1 exerts its effects by specifically binding to its receptor CXCR2, has the function of chemotactic neutrophils, and is also involved in many important physiological and pathological reactions, such as regulating inflammatory reactions and immune responses. Many studies have reported that CXCL1 is highly expressed in solid tumors such as melanoma, ovarian cancer, colorectal cancer, and gastric cancer, and is closely related to tumor progression. CXCL5 (CXC chemokine 5, ENA-78), as a powerful inducer and inflammatory mediator of neutrophils, can mediate various types of inflammatory reactions and promote cell migration, proliferation and other life activities. CXCL5 can mediate the progression of digestive tract tumors such as colorectal cancer, gastric cancer, esophageal squamous cell carcinoma and pancreatic cancer, as well as the interaction between tumor cells and the microenvironment. Both tumor cells and microenvironmental cell components can release CXCL11 (CXC motif chemokine 11, I-TAC), which in turn mediates the directional movement of tumor cells, participates in the interaction between tumor cells and microenvironmental cell components, and mediates tumor cell migration out of blood vessels, immune escape, and metastasis. In oncology research, CXCL11 can mediate the progression of various solid tumors by activating multiple signaling pathways.

[0004] Many tumors have an insidious onset, lack of clinically specific manifestations, and cannot be systematically evaluated through a combination of multiple testing methods, which leads to the late stage of cancer patients when they seek medical treatment and poor prognosis. Plasma testing can directly detect biomarkers through blood samples obtained through minimally invasive methods. It is convenient for testing, low cost, high efficiency, and has excellent application prospects. However, so far, there are no biomarkers that can be used for non-invasive testing to diagnose whether esophageal squamous cell carcinoma has metastasized. Summary of the invention

[0005] The present invention aims to solve at least one of the technical problems existing in the related art, and provides an application of CXCL1, CXCL5 and CXCL11 chemokines in preparing a kit for diagnosing metastasis of esophageal squamous cell carcinoma. Specifically, the present invention provides the following technical solutions:

[0006] In a first aspect, the present invention provides a use of CXCL1, CXCL5 and CXCL11 chemokines in the preparation of a kit for diagnosing metastasis of esophageal squamous cell carcinoma, wherein the kit comprises reagents for detecting the concentrations of CXCL1, CXCL5 and CXCL11 chemokines in the plasma of esophageal cancer patients.

[0007] In one embodiment, the esophageal squamous cell carcinoma metastasis is lymphatic metastasis. BRIEF DESCRIPTION OF THE DRAWINGS

[0008] The accompanying drawings are used to provide a further understanding of the present invention and constitute a part of the specification. Together with the embodiments of the present invention, they are used to explain the present invention and do not constitute a limitation of the present invention. In the accompanying drawings:

[0009] Figure 1 It is the ROC curve diagram. DETAILED DESCRIPTION

[0010] The preferred embodiments of the present invention are described below in conjunction with the accompanying drawings. It should be understood that the preferred embodiments described herein are only used to illustrate and explain the present invention, and are not used to limit the present invention.

[0011] Example 1: CXCL1, CXCL5 and CXCL11 chemokines are highly expressed in the plasma of patients with esophageal squamous cell carcinoma metastasis

[0012] 1. Sample extraction:

[0013] A total of 32 patients with non-metastatic esophageal squamous cell carcinoma and 57 patients with lymphatic metastasis of esophageal squamous cell carcinoma (from Peking University Cancer Hospital, all of whom had signed informed consent) were enrolled, and venous blood and plasma samples were drawn from the patients.

[0014] Add samples to CXCL1 ELISA 96-well plate (CUSABIO, catalog number: CSB-E09150h), CXCL5 ELISA 96-well plate (CUSABIO, catalog number: CSB-E08178h) and CXCL11 ELISA 96-well plate (CUSABIO, catalog number: CSB-E09023h) in sequence, seal and incubate at 37 °C for 1 hour. After incubation, discard the samples and wash them repeatedly with cleaning solution 5 times. Add biotin-linked CXCL1, CXCL5 and CXCL11 antibodies in the ELISA kit to the 96-well plate (100 μL / well), shake and incubate at room temperature for 1 hour, wash repeatedly with cleaning solution and discard. Add reaction substrate to each well, add reaction stop solution at room temperature in the dark, and measure the absorbance value at 450 nm using an ELISA reader.

[0015] The expression differences of chemokines CXCL1, CXCL5 and CXCL11 in the plasma samples of 57 metastatic and 32 non-metastatic groups were compared (Table 1), and the Spearman test was used to observe the correlation between the expression of CXCL1, CXCL5 and CXCL11 in the plasma of the 57 metastatic groups.

[0016] Table 1

[0017]

[0018]

[0019]

[0020]

[0021] Based on the above sample set, a ROC curve was created to determine the accuracy of CXCL1, CXCL5, and CXCL11 as plasma markers. The model training data was used to calculate the true positive rate and false positive rate at each possible threshold. The ROC curve was drawn with the false positive rate as the horizontal axis and the true positive rate as the vertical axis. Each point represents a specific threshold, and these points are connected to form a ROC curve. The AUC value was calculated to quantify the performance of the model. According to the ROC curve results ( Figure 1), when used alone, the diagnostic power AUC for CXCL1 was 0.911, the diagnostic power AUC for CXCL5 was 0.832, and the diagnostic power AUC for CXCL11 was 0.885. When used in combination, the combined diagnostic power AUC of CXCL1 and CXCL5 was 0.987, the combined diagnostic power AUC of CXCL5 and CXCL11 was 0.952, and the combined diagnostic power AUC of CXCL1 and CXCL11 was 0.979. When CXCL1, CXCL5, and CXCL11 were combined, the diagnostic power AUC was as high as 0.997. The diagnostic critical values ​​of the three biomarkers were CXCL1=169.438 pg / mL, CXCL5=716.753 pg / mL, and CXCL11=486.766 pg / mL, respectively. When the measured values ​​of CXCL1, CXCL5, and CXCL11 in the sample were all greater than the diagnostic critical values, the patient was diagnosed with esophageal squamous cell carcinoma metastasis.

[0022] Example 2 Validation of the effectiveness of CXCL1, CXCL5 and CXCL11 as biomarkers for detecting metastasis of esophageal squamous cell carcinoma

[0023] In order to verify the effectiveness of CXCL1, CXCL5 and CXCL11 as biomarkers for detecting esophageal squamous cell carcinoma metastasis, we re-enrolled 15 patients with esophageal squamous cell carcinoma (as a validation set, from Peking University Cancer Hospital, all of whom had signed informed consent), analyzed the samples of the patients in the validation set using the biomarkers and diagnostic cutoff values ​​described in Example 1 above, and compared the analysis results with the clinical expert judgment results (Table 2):

[0024] Table 2

[0025]

[0026] As can be seen from the data in Table 2, CXCL1, CXCL5 and CXCL11 have extremely high accuracy as biomarkers for detecting esophageal squamous cell carcinoma metastasis.

[0027] Obviously, those skilled in the art can make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if these modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include these modifications and variations.

Claims

1. A use of CXCL1, CXCL5 and CXCL11 in the preparation of a kit for diagnosing metastasis of esophageal squamous cell carcinoma, characterized in that: The kit comprises reagents for detecting the concentrations of CXCL1, CXCL5 and CXCL11 in the plasma of patients with esophageal cancer, wherein the metastasis of the esophageal squamous cell carcinoma is lymphatic metastasis.

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