Application of serum EPSTI1 in preparation of products for diagnosing liver cancer and evaluating whether liver cancer patients are resistant to revatinib

Abstract

The invention relates to the field of biological medicines, and discloses application of serum EPSTI1 in preparation of a product for diagnosing liver cancer and evaluating whether a liver cancer patient is resistant to a revatinib or not. The invention provides a detection means which is convenient and rapid in sampling, small in risk and relatively low in cost, the content of EPSTI1 in serum is detected through an enzyme-linked immunosorbent assay, a liver cancer patient and a non-liver cancer patient can be effectively distinguished, and a lovatinib drug-resistant liver cancer patient and a non-drug-resistant liver cancer patient can be effectively distinguished. Prognosis analysis shows that the serum EPSTI1 can be used as a marker for evaluating the prognosis state of the liver cancer patient taking the lenvatinib, and has important significance in improving the survival rate of the liver cancer patient.

Description

Technical Field

[0001] This application relates to the field of biomedicine, and more specifically, to the use of serum EPSTI1 in the preparation of products for the diagnosis of liver cancer and the assessment of whether liver cancer patients are resistant to lenvatinib. Background Technology

[0002] Hepatocellular carcinoma (HCC) is a malignant tumor originating from liver cells. It ranks sixth in incidence among malignant tumors worldwide and is the third leading cause of cancer-related death. The incidence of HCC in my country is higher than in other regions globally, ranking fourth among all cancers, and has become a serious public health problem. For early-stage HCC patients, local ablation and surgical resection are commonly used for radical treatment; for mid-to-late-stage patients, radiofrequency ablation, hepatic artery chemoembolization, and liver transplantation are used. In recent years, with the continuous improvement of tumor gene mapping, more and more kinase inhibitors have been applied to clinical cancer treatment, significantly improving patient prognosis and survival.

[0003] Lenvatinib is a multi-receptor tyrosine kinase inhibitor approved in China for first-line treatment of intermediate-to-advanced hepatocellular carcinoma (HCC). Its efficacy in HCC patients is comparable to sorafenib. Studies have reported that HCC patients in China treated with lenvatinib have better disease-free survival and overall survival than those treated with sorafenib. However, 50-80% of patients develop resistance after treatment, ultimately leading to adverse events such as tumor progression and distant metastasis. Therefore, identifying novel biomarkers indicating lenvatinib sensitivity is beneficial for clinical classification and precision treatment of patients with intermediate-to-advanced HCC, thereby improving patient survival rates.

[0004] Epithelial-mesenchymal interaction protein 1 (EPSTI1) was initially identified as a stromal fibroblast-inducing gene in human breast cancer and is significantly elevated in tumor tissues of breast cancer patients. In normal cells, EPSTI1 is thought to be associated with cell adhesion and immune cell activation. Subsequent studies have found that EPSTI1 is aberrantly expressed in various tumor types and is also associated with tumor survival, invasion, and metastasis. It is believed that the biological functions of EPSTI1 vary significantly depending on cell type. However, the expression and clinical value of serum EPSTI1 in liver cancer patients have not yet been reported. Summary of the Invention

[0005] The purpose of this invention is to overcome the technical problem that most existing liver cancer drugs lack effective drug sensitivity biomarkers, and to explore a rapid, convenient and effective serological biomarker for predicting the prognosis of liver cancer patients taking lenvatinib and improving patient survival.

[0006] To achieve the above-mentioned objectives, this application provides the following technical solution:

[0007] In the first aspect, this application provides the application of a reagent for detecting serum EPSTI1 in the preparation of liver cancer diagnostic products.

[0008] Furthermore, the reagent is a blood test reagent, and the test product is a test kit.

[0009] Furthermore, the kit is an enzyme-linked immunosorbent assay (ELISA) kit.

[0010] Furthermore, the reagent for detecting serum EPSTI1 includes an EPSTI1 antibody.

[0011] Secondly, the application of reagents for detecting serum EPSTI1 in the preparation of products for assessing whether liver cancer patients are resistant to lenvatinib.

[0012] Furthermore, the reagent is a blood test reagent, and the test product is a test kit.

[0013] Furthermore, the kit is an enzyme-linked immunosorbent assay (ELISA) kit.

[0014] Furthermore, the reagent for detecting serum EPSTI1 includes an EPSTI1 antibody.

[0015] In summary, this application has the following beneficial effects: The serum EPSTI1 provided by this invention serves as a biomarker for liver cancer, offering a convenient, rapid, low-risk, and cost-effective detection method. This invention detects serum EPSTI1 levels using an enzyme-linked immunosorbent assay (ELISA). Results showed significant differences in serum EPSTI1 expression levels among liver cancer patients, chronic hepatitis B (CHB) patients, and healthy donors (HD), effectively distinguishing between lenvatinib-resistant and non-resistant liver cancer patients. Prognostic analysis indicates that serum EPSTI1 can serve as a biomarker for assessing the prognostic status of liver cancer patients taking lenvatinib, and is of significant importance in improving the survival rate of liver cancer patients. Attached Figure Description

[0016] Figure 1 Training focuses on serum EPSTI1 to differentiate between hepatocellular carcinoma (HCC) and non-HCC patients / Lenvatinib-resistant and non-resistant HCC patients; Figure 2 Serum EPSTI1 can assess the prognosis of liver cancer patients taking lenvatinib; Figure 3 : To validate the use of serum EPSTI1 to differentiate between hepatocellular carcinoma (HCC) and non-HCC patients, and between lenvatinib-resistant and non-resistant HCC patients. Detailed Implementation

[0017] The technical solutions and effects of this application will be further described in detail below with reference to embodiments and accompanying drawings. It should be understood that the specific embodiments described herein are merely illustrative of the invention and not intended to limit it. Furthermore, it should be noted that, for ease of description, the accompanying drawings show only the parts relevant to the invention, not the entire structure.

[0018] Lenvatinib is a first-line drug for treating intermediate-to-advanced hepatocellular carcinoma (HCC), but 50-80% of patients develop resistance after treatment, ultimately leading to adverse events such as tumor progression and distant metastasis, resulting in a low five-year survival rate for patients with intermediate-to-advanced HCC. Therefore, identifying biomarkers that indicate lenvatinib sensitivity is beneficial for clinical classification and precise treatment of patients with intermediate-to-advanced HCC, and helps improve patient prognosis and survival. Serum EPSTI1 levels are high in lenvatinib-resistant HCC patients and can effectively differentiate patient prognostic status, suggesting that it may become a novel biomarker for predicting lenvatinib sensitivity in HCC.

[0019] Therefore, this invention uses enzyme-linked immunosorbent assay (ELISA) to detect serum EPSTI1 levels. The results showed significant differences in serum EPSTI1 expression levels among patients with hepatocellular carcinoma (HCC), patients with chronic hepatitis B (CHB), and healthy donors (HD), and it could effectively distinguish between lenvatinib-resistant and non-resistant HCC patients. Prognostic analysis indicated that serum EPSTI1 can serve as a biomarker for assessing the prognostic status of HCC patients taking lenvatinib, and is of great significance for improving the survival rate of HCC patients. Example 1 1. Serum sample collection Serum samples were collected from patients at the Department of Laboratory Medicine, Zhongshan Hospital Affiliated to Fudan University, between 2021 and 2022. Patients were divided into four groups: hepatocellular carcinoma (HCC) patients treated with lenvatinib (HCC, n=50), chronic hepatitis B patients (CHB, n=50), and healthy controls (HD, n=50). Based on tumor progression after lenvatinib administration, HCC patients were further divided into a response group (R, n=25) and a non-response group (NR, n=25). Progression-free survival (PFS) was defined as the time from the start of treatment to disease progression or death from any cause. Overall survival (OS) was defined as the time from the start of treatment to death from any cause. Survival data were collected up to June 2023. All serum samples were collected before treatment, aliquoted, and stored at -80°C.

[0020] 2. Enzyme-linked immunosorbent assay (ELISA) The serum EPSTI1 content was detected using an ELISA kit (CLOUD-CLONE CORP. WUHAN, SEG200Hu). The experimental procedure is briefly described as follows: (1) Add serum sample to the microwells of the solid-phase carrier coated with EPSTI1 antibody and incubate at 37°C for 1 hour; (2) Discard the liquid in the wells, add biotinylated EPSTI1 detection antibody, and incubate at 37°C for 1 hour; (3) Discard the liquid in the wells, wash the plate 3 times to remove unbound biotinylated antibody, add horseradish peroxidase-labeled avidin, and incubate at 37°C for 30 minutes; (4) Discard the liquid in the wells, wash the plate 5 times, add chromogenic reagent TMB, and develop color at 37°C in the dark for 15 minutes; (5) Add acidic stop solution, measure the absorbance at 450 nm using an ELISA reader, and calculate the EPSTI1 concentration in the serum sample.

[0021] 3. Statistical Analysis Data were statistically analyzed using SPSS and Graphpad Prism software. The Student t-test was used to assess whether there was a significant difference between two groups with homogeneous variances. The Mann-Whitney U test was used to assess whether there was a significant difference between two groups with unequal variances. PFS and OS in liver cancer patients were assessed by plotting Kaplan-Meier survival curves. The diagnostic efficacy of serum EPSTI1 was evaluated using receiver operating characteristic (ROC) curves. Sensitivity and specificity were recorded when the Youden index (sensitivity + specificity - 1) was maximized. A p-value less than 0.05 was considered statistically significant.

[0022] 4. Evaluate the clinical value of serum EPSTI1 as a prognostic biomarker for liver cancer patients taking lenvatinib. We measured serum EPSTI1 levels using ELISA, and the results showed that serum EPSTI1 levels in hepatocellular carcinoma (HCC) patients were significantly higher than those in patients with chronic hepatocellular carcinoma (CHB) and hepatocellular carcinoma (HD), and could effectively distinguish HCC from non-HCC patients. Figure 1 AB). In hepatocellular carcinoma patients treated with lenvatinib, those who developed resistance had higher serum EPSTI1 levels, which can effectively differentiate between lenvatinib-resistant and non-resistant hepatocellular carcinoma patients. Figure 1 CD). We further analyzed the prognostic role of EPSTI1 in a cohort of liver cancer patients taking lenvatinib. Patients were divided into high / low serum EPSTI1 groups based on median test results. The results showed that patients with high serum EPSTI1 levels had significantly shorter PFS and OS than those with low serum EPSTI1 levels. Figure 2 (AB). The above data suggest that serum EPSTI1 may be an independent predictor of prognosis for liver cancer patients taking lenvatinib.

[0023] Example 2 1. Serum sample collection Serum samples were collected from patients at the Department of Laboratory Medicine, Zhongshan Hospital Affiliated to Fudan University, between 2022 and 2023. Patients were divided into four groups: hepatocellular carcinoma (HCC) patients treated with lenvatinib (HCC, n=30), chronic hepatitis B patients (CHB, n=20), and healthy controls (HD, n=20). Based on tumor progression after lenvatinib administration, HCC patients were further divided into a response group (R, n=13) and a non-response group (NR, n=17). All serum samples were collected prior to treatment, aliquoted, and stored at -80°C.

[0024] 2. Enzyme-linked immunosorbent assay (ELISA) Same as Example 1 3. Statistical Analysis Same as Example 1 4. Validation set to validate the clinical value of serum EPSTI1 In the validation set, serum EPSTI1 levels in hepatocellular carcinoma patients were also effective in distinguishing between hepatocellular carcinoma and non-hepatocellular carcinoma patients. Figure 3 AB). In hepatocellular carcinoma patients treated with lenvatinib, those who developed resistance had higher serum EPSTI1 levels, which can effectively differentiate between lenvatinib-resistant and non-resistant hepatocellular carcinoma patients. Figure 3 CD).

[0025] This specific embodiment is merely an explanation of this application and is not intended to limit it. After reading this specification, those skilled in the art can make modifications to this embodiment without contributing any inventive step, but such modifications are protected by patent law as long as they fall within the scope of the claims of this application.

Claims

1. Application of reagents for detecting serum EPSTI1 in the preparation of liver cancer diagnostic products.

2. The application according to claim 1, characterized in that, The reagent is a blood test reagent, and the test product is a test kit.

3. The application according to claim 2, characterized in that, The kit is an enzyme-linked immunosorbent assay (ELISA) kit.

4. The application according to claim 2, characterized in that, The reagent for detecting serum EPSTI1 includes an EPSTI1 antibody.

5. Application of reagents for detecting serum EPSTI1 in the preparation of products for assessing whether liver cancer patients are resistant to lenvatinib.

6. The application according to claim 5, characterized in that, The reagent is a blood test reagent, and the test product is a test kit.

7. The application according to claim 6, characterized in that, The kit is an enzyme-linked immunosorbent assay (ELISA) kit.

8. The application according to claim 6, characterized in that, The reagent for detecting serum EPSTI1 includes an EPSTI1 antibody.

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