Method for constructing nasopharyngeal carcinoma prognosis model by jointly detecting tumor-related markers

Abstract

The present application relates to a method for constructing a nasopharyngeal carcinoma prognosis model by jointly detecting tumor-related markers, comprising the following steps: Step one, obtaining nasopharyngeal carcinoma tissue sections of a patient; Step two, obtaining the nasopharyngeal carcinoma tissue sections can represent the tumor-related infiltrating lymphocyte marker CD4 + T, CD8 + T and a group of markers related to angiogenesis VEGF-A, VEGFR-2, and calculating the immunohistochemical scores of the markers CD4+T, CD8+T, VEGF-A and VEGFR-2; Step three, calculating the co-expression score of the angiogenesis-related factor and the tumor infiltrating lymphocyte of the patient according to the immunohistochemical scores. The present application provides a new and reliable molecular marker model for the prognosis of nasopharyngeal carcinoma patients, and provides a new target for the use of immunotherapy in nasopharyngeal carcinoma.

Description

Technical Field

[0001] This invention relates to the medical field, specifically to a method for constructing a prognostic model for nasopharyngeal carcinoma by combining the detection of tumor-related biomarkers. Background Technology

[0002] Because nasopharyngeal carcinoma (NPC) often occurs in hidden locations, its clinical symptoms are atypical and it is difficult to detect in its early stages. Therefore, over 70% of NPC patients are diagnosed at a locally advanced stage. While the development of intensity-modulated radiotherapy (IMRT) and combined chemoradiotherapy have significantly improved local control rates, approximately 30-40% of locally advanced NPC patients still experience distant metastasis after receiving standardized NPC treatment. Currently, there is no reliable and effective treatment for recurrent or metastatic NPC. Treatment of NPC is still based on clinical staging according to the tumor's primary lesion-lymph node metastasis-distant metastasis (TNM) classification, as determined by the International Union Against Cancer (UICC) and the American Joint Committee on Cancer (AJCC). However, even among NPC patients with the same TMN stage and receiving similar treatments, clinical outcomes vary considerably. Therefore, there is an urgent need to find a new classification method to better screen NPC patients at high risk of recurrence and metastasis, enabling early intervention and individualized treatment, facilitating clinical management of NPC, and further improving patient survival. Summary of the Invention

[0003] To overcome the shortcomings of existing prognostic methods for nasopharyngeal carcinoma patients, and given the advent of precision medicine, immunotherapy combined with targeted therapy has shown superiority in various advanced recurrent and metastatic tumors. The technical problem this invention aims to solve is to provide a method for constructing a prognostic model for nasopharyngeal carcinoma by jointly detecting tumor-related biomarkers. This method identifies a set of biomarkers that can represent acquired immune-related factors (CD4+). + T, CD8 + T) and angiogenesis-related markers (VEGF-A, VEGFR-2). The technical solution of this invention to solve the above-mentioned technical problems is as follows: A method for constructing a prognostic model for nasopharyngeal carcinoma by jointly detecting tumor-related biomarkers, comprising the following steps: Step 1: Obtain a tissue section of the patient's nasopharyngeal carcinoma. Step two involves obtaining nasopharyngeal carcinoma tissue sections, which can represent CD4 markers of tumor-associated infiltrating lymphocytes. + T, CD8 + T and a group of biomarkers that can represent angiogenesis-related markers, VEGF-A and VEGFR-2, and calculate the immunohistochemical scores of biomarkers CD4+T, CD8+T, VEGF-A and VEGFR-2; Step 3: Calculate the co-expression score of angiogenesis-related factors and tumor-infiltrating lymphocytes based on the immunohistochemical score; Step four: Obtain the patient's risk score based on the co-expression score of angiogenesis-related factors and tumor-infiltrating lymphocytes, as well as the TNM stage.

[0004] The beneficial effects of this invention are: it provides a new and reliable molecular marker model for predicting the prognosis of nasopharyngeal carcinoma patients and provides a new target for the application of immunotherapy in nasopharyngeal carcinoma.

[0005] Based on the above technical solution, the present invention can be further improved as follows.

[0006] Furthermore, in step two, the immunohistochemical scores of the biomarkers CD4+T, CD8+T, VEGF-A, and VEGFR-2 are calculated using the following methods: (1) The staining intensity of CD4+T, CD8+T, VEGF-A and VEGFR-2 positive cells is negative, weakly positive, moderately strong and strongly positive respectively, and is scored as 0, 1, 2 and 3 points respectively. The percentage of CD4+T, CD8+T, VEGF-A and VEGFR-2 positive cells is less than 25%, between 25% and 50%, between 51% and 75% and greater than 75% respectively, and is scored as 1, 2, 3 and 4 points respectively. (2) The immunohistochemical score is obtained by multiplying the score of staining intensity of positive cells for markers CD4+T, CD8+T, VEGF-A and VEGFR-2 by the score of the percentage of positive cells.

[0007] The beneficial effect of the above-mentioned further technical solution is to obtain an immunohistochemical score.

[0008] Furthermore, in step three, the co-expression score of angiogenesis-related factors and tumor-infiltrating lymphocytes in the patient is calculated using the following method: CO V+TIL score = H-score VEGFA + H-score VEGFR2 +[12-(H-score CD4 )) + [(12- H-score CD8 )]; Among them, CO V+TIL The score represents the co-expression score of angiogenesis-related factors and tumor-infiltrating lymphocytes, H-score. VEGFA The H-score is used to assess the immunohistochemical score of the biomarker VEGF-A. VEGFR2The immunohistochemical score for the biomarker VEGFR-2, H-score CD4 The immunohistochemical score of the CD4+ T marker, H-score CD8 Immunohistochemical score for the biomarker CD8+T.

[0009] The beneficial effect of the above-mentioned further technical solution is that it obtains a score for the co-expression of angiogenesis-related factors and tumor-infiltrating lymphocytes.

[0010] Furthermore, it also includes step four: obtaining the patient's risk score based on the co-expression score of angiogenesis-related factors and tumor-infiltrating lymphocytes, as well as the TNM stage.

[0011] Furthermore, based on the patients' angiogenesis-related factor and tumor-infiltrating lymphocyte co-expression scores and TNM staging, the patients' risk scores were obtained using the following method: patients with angiogenesis-related factor and tumor-infiltrating lymphocyte co-expression scores less than 28 were assigned to the low group, and those with scores greater than 28 were assigned to the high group, with the low group assigned 0 points and the high group assigned 1 point. (2) Patients in TNM stage 1-3 are given 0 points, and those in TNM stage 4 are given 1 point; (3) Add the scores from step (1) and step (2) together to obtain the patient’s risk score.

[0012] The beneficial effects of the above-mentioned further technical solutions are: the co-expression score of angiogenesis-related factors and tumor-infiltrating lymphocytes is combined with TNM staging. Attached Figure Description

[0013] Figure 1 This is a representative image of immunohistochemical staining in nasopharyngeal carcinoma tissue, showing low expression of VEGF-A (expression scale bar at 50 μm). Figure 2 This is a representative image of immunohistochemical staining in nasopharyngeal carcinoma tissue, showing low expression of VEGFR-2 (expression scale bar at 50 μm). Figure 3 This is a representative image of immunohistochemical staining in nasopharyngeal carcinoma tissue, CD4 + Low expression of T (expression scale bar is 50 μm); Figure 4 This is a representative image of immunohistochemical staining in nasopharyngeal carcinoma tissue, CD8 + Low expression of T (expression scale bar is 50 μm); Figure 5 This is a representative image of immunohistochemical staining in nasopharyngeal carcinoma tissue, showing high expression of VEGF-A (expression scale bar is 50 μm). Figure 6This is a representative image of immunohistochemical staining in nasopharyngeal carcinoma tissue, showing high expression of VEGFR-2 (expression scale bar is 50 μm). Figure 7 This is a representative image of immunohistochemical staining in nasopharyngeal carcinoma tissue, CD4 + High expression of T (expression scale bar is 50 μm); Figure 8 This is a representative image of immunohistochemical staining in nasopharyngeal carcinoma tissue, CD8 + High expression of T (expression scale bar is 50 μm); Figure 9 To determine the risk score threshold for the co-expression of angiogenesis and immune lymphoid infiltration using X-tile software; Figure 10 Overall survival (OS) was plotted for the training cohort based on the co-expression score of angiogenesis-related markers and immune infiltration markers. Figure 11 Progression-free survival (DFS) curves plotted for the training cohort based on the co-expression score of angiogenesis-related markers and immune infiltration markers. Figure 12 Local recurrence-free scores (LRRFS) were plotted in the training cohort based on the co-expression scores of angiogenesis-related markers and immune infiltration markers. Figure 13 The survival curves (DMFS) for no and distant metastasis in the training cohort were plotted based on the co-expression scores of angiogenesis-related markers and immune infiltration markers. Figure 14 To validate the overall survival (OS) plotted in the cohort based on the co-expression score of angiogenesis-related markers and immune infiltration markers; Figure 15 To validate the progression-free survival (DFS) curves plotted based on the co-expression scores of angiogenesis-related markers and immune infiltration markers in the cohort. Figure 16 To validate the local recurrence-free score (LRRFS) plotted based on the co-expression score of angiogenesis-related markers and immune infiltration markers in the cohort; Figure 17 To validate the survival curves (DMFS) of no metastasis and distant metastasis in the cohort based on the co-expression scores of angiogenesis-related markers and immune infiltration markers. Figure 18 CO V+TIL Overall survival (OS) was compared between the risk score, which combines the risk score with the TNM staging, and the recipient operating characteristic curve of the TNM staging. Figure 19 CO V+TILProgression-free survival (DFS) curves comparing the risk score, which combines the risk score with the TNM staging, and the recipient operating characteristic curve of the TNM staging. Figure 20 CO V+TIL Local recurrence-free (LRRFS) comparison between the risk score combined with TNM staging and the recipient operating characteristic curve of TNM staging. Figure 21 CO V+TIL Risk score combining TNM staging and receiver operating characteristic curves of TNM staging with no distant metastasis (DMFS). Detailed Implementation

[0014] The principles and features of the present invention are described below with reference to the accompanying drawings. The examples given are only for explaining the present invention and are not intended to limit the scope of the present invention.

[0015] Example 1 A method for constructing a prognostic model for nasopharyngeal carcinoma by jointly detecting tumor-related biomarkers includes the following steps: Step 1: Obtain a tissue section of the patient's nasopharyngeal carcinoma.

[0016] Step two involves obtaining nasopharyngeal carcinoma tissue sections, which can represent CD4 markers of tumor-associated infiltrating lymphocytes. + T, CD8 + T and a group of biomarkers that can represent angiogenesis, VEGF-A and VEGFR-2, were used to calculate the immunohistochemical scores of the biomarkers CD4+T, CD8+T, VEGF-A and VEGFR-2 using the following method: (1) The staining intensity of CD4+T, CD8+T, VEGF-A and VEGFR-2 positive cells is negative, weakly positive, moderately strong and strongly positive respectively, and is scored as 0, 1, 2 and 3 points respectively. The percentage of CD4+T, CD8+T, VEGF-A and VEGFR-2 positive cells is less than 25%, between 25% and 50%, between 51% and 75% and greater than 75% respectively, and is scored as 1, 2, 3 and 4 points respectively. (2) The immunohistochemical score is obtained by multiplying the score of staining intensity of positive cells for markers CD4+T, CD8+T, VEGF-A and VEGFR-2 by the score of the percentage of positive cells.

[0017] Step 3: Based on the immunohistochemical score, the co-expression score of angiogenesis-related factors and tumor-infiltrating lymphocytes in the patient is calculated using the following method: CO V+TIL score = H-scoreVEGFA + H-score VEGFR2 +[12-(H-score CD4 )) + [(12- H-score CD8 )]; Among them, CO V+TIL The score represents the co-expression score of angiogenesis-related factors and tumor-infiltrating lymphocytes, H-score. VEGFA The H-score is used to assess the immunohistochemical score of the biomarker VEGF-A. VEGFR2 The immunohistochemical score for the biomarker VEGFR-2, H-score CD4 The immunohistochemical score of the CD4+ T marker, H-score CD8 Immunohistochemical score for the biomarker CD8+T.

[0018] The following experimental comparisons illustrate the beneficial effects of this invention: Experiment 1 Step 1: Collected 238 nasopharyngeal carcinoma tissue sections from two medical institutions, divided into a training cohort (n=124) and a validation cohort (n=114). Selected a set of markers representing tumor-associated infiltrating lymphocytes, CD4. + T, CD8 + T and VEGF-A and VEGFR-2, which can represent a group of markers related to angiogenesis, were measured to detect their expression levels.

[0019] Step 2: Immunohistochemical scoring was used to represent the expression levels of target genes. Representative and viable tissue sections were scored using an optical microscope. Angiogenesis markers VEGF-A and VEGFR-2 were positively expressed in tumor cell membranes and cytoplasm, and CD4+ was observed in lymphoinfiltrating cells. + T and CD8 +T cells were located in the tumor stroma and were distributed in brownish-yellow or brownish-red granular form. Two pathologists independently evaluated the experimental results in a double-blind manner. Under an optical microscope, the entire section was observed at high magnification (10*40) and the product of the staining intensity of positive cells and the percentage of positive cells was used as the judgment index. The staining intensity of positive cells for the markers CD4+T, CD8+T, VEGF-A, and VEGFR-2 was scored as 0, 1, 2, and 3 points, respectively, for negative, weakly positive, moderate, and strongly positive staining. The percentage of positive cells for the markers CD4+T, CD8+T, VEGF-A, and VEGFR-2 was scored as 1, 2, 3, and 4 points, respectively, for percentages less than 25%, between 25% and 50%, between 51% and 75%, and greater than 75%. Finally, the immunohistochemical scores of the biomarkers CD4+T, CD8+T, VEGF-A, and VEGFR-2 were obtained from 0 to 12 points. Those with immunohistochemical scores of 0-7 points were assigned to the low expression group, and those with scores of 8-12 points were assigned to the high expression group. Figure 1-8 (As shown).

[0020] Step 3: A model for predicting tumor prognosis was established based on immunohistochemical scores of angiogenesis markers and immunohistochemical scores of molecular markers of tumor immune infiltrating cells. Numerous studies have confirmed that nasopharyngeal carcinoma patients with high expression of angiogenesis factors have a poor prognosis, while those with high expression of tumor-infiltrating lymphocytes have a better prognosis.

[0021] Step 4: Existing research lacks a calculation method that can simultaneously unify multiple biomarkers with opposing prognostic predictions for prognostic assessment. This invention creates a formula that unifies their scores for prognostic prediction. Since the above uses immunohistochemical scoring to reflect the expression level of target biomarkers, the total score is 12. Subtracting the immunohistochemical scores of prognostic positively correlated factors from 12 converts these factors into negatively correlated ones. The formula is obtained by adding 12 to the immunohistochemical scores of prognostic positively correlated biomarkers (i.e., co-expression risk scores) of genes with high expression and poor prognosis (H-Score) plus the immunohistochemical scores of genes with low expression and poor prognosis (H-Score), resulting in the following formula: CO V+TIL score = H - score Hpoor +[12-(H-score Lpoor The formula after incorporating the target gene from this invention is as follows: CO V+TIL score = H-score VEGFA + H-score VEGFR2 +[12-(H-score CD4 )) + [(12-H-scoreCD8 Ultimately, the CO for each patient can be obtained. V+TIL score, CO of patients in the training cohort V+TIL The score range is 6-45 (median, 26), validating the CO of patients in the cohort. V+TIL The score range is 4-46 (median, 23). CO V+TIL A high score indicates high expression of vascular-related factors and low expression of tumor immune infiltration lymphokines. V+TIL A low score indicates low expression of vascular-related factors and low expression of tumor immune infiltration lymphokines.

[0022] Step 5: To separate the low-risk and high-risk co-expression groups, this invention used X-tile software (a software developed by Yale University to select the optimal cut-off value in survival data) to determine the optimal cut-off value for the co-expression risk score in 124 patients with complete prognostic information in the training cohort. Figure 9 As shown, the cutoff value for the co-expressed risk score is equal to 28, which is the optimal cutoff value selected by the X-tile software. The optimal cutoff value (28) was generated using the X-tile software to stratify patients of different risks in the training cohort. In the training cohort, there were a total of 49 (40%) patients (CO). v+TIL The high-risk group (score: 28-45) consisted of 75 (60%) patients (CO). v+TIL Patients with scores of 6-27 were classified as low-risk. The 5-year overall survival, progression-free survival, and local recurrence-free survival rates of high-risk patients were significantly different from those of the low-risk group, with OS of (62.4% vs. 95.8%, HR: 9.08, 95% CI: 2.65–31.12), respectively. p <0.001), PFS (36.5% vs .88.1%, HR: 6.52, 95%CI: 3.08–13.77, p <0.001); LRRFS (71.3% vs.92.7, HR: 3.26, 95%CI: 1.22–8.72, p =0.015); and DMFS (52.6% vs. 92.6%, HR: 8.02, 95% CI: 3.03–21.23, p <0.001)( Figure 10-13 (As shown).

[0023] Multivariate Cox regression analysis was performed on the training queue, and the results showed that CO V+TILThe score remained significant for OS, PFS, LRRFS, and DMFS in the training queue (OS: HR: 0.120, 95% CI: 0.036 -0.422). p = 0.007; PFS: HR: 0.158, 95% CI 0.075 - 0.334, p < 0.001, LRRFS: HR: 0.326, 95% CI: 0.122 -0.874, p = 0.026; DMFS: HR: 0.132, 95% CI: 0.50-0.349, p < 0.001). Furthermore, multivariate Cox analysis showed that CO V+TIL Score is an independent prognostic factor for nasopharyngeal carcinoma.

[0024] Step 6: Validate CO in the validation group V+TIL Survival analysis validation of the risk scoring model To verify CO V+TIL The accuracy of the risk score in survival prognostic analysis was assessed by classifying patients in the validation cohort into high CO groups using the same optimal cut-off value (cut-of-value 28) as in the training cohort. V+TIL score and low CO V+TIL score. Consistent with the training queue results, CO V+TIL Patients with high risk scores were more likely to develop CO V+TIL The low-risk group showed significant differences in OS, PFS, and LRRFS. OS (46.8% vs. 83.4%, HR: 3.42, 95% CI: 1.77–6.61) p <0.001); PFS (45.9% vs. 81.2%, HR: 3.22, 95%CI: 1.71–6.06, p <0.001); LRRFS (74.6% vs. 87.5%, HR: 3.22, 95%CI: 1.16–8.93, p =0.027); andDMFS (79.2% vs. 93.2%, HR: 2.22, 95%CI: 0.91-5.39, p =0.086) Figure 14-17 (As shown).

[0025] Cox regression analysis was performed on the validation group, and the results showed that CO V+TILThe score remained significant for OS, PFS, LRRFS, and DMFS in the training queue (OS: HR: 0.115, 95% CI: 0.049 -0.271). p < 0.001; PFS: HR: 1.173, 95% CI 0.80 - 0.374, p < 0.001, LRRFS: HR: 0.279, 95% CI: 0.091 -0.857, p = 0.026; DMFS: HR 0.325, 95% CI: 0.121-0.870, p =0.025). Furthermore, multivariate Cox analysis showed that CO V+TIL The score remains a stable independent prognostic factor.

[0026] Experiment 2 Application of CO V+TIL The efficacy of induction chemotherapy was assessed using a risk score model. A total of 238 nasopharyngeal carcinoma patients were included, of whom 135 received platinum-based induction chemotherapy and had post-chemoradiographic evaluations. According to CO... V+TIL Using a risk score, 135 patients were divided into a low-risk group (n=83, 61%) and a high-risk group (n=52, 39%). The overall response rate (ORR) in the low-risk group was 78% (65 / 83); 22% (18 / 83) of these patients reported stable disease (SD). In the high-risk group, the ORR was 65% (34 / 52); 35% (18 / 52) of these patients reported both SD and progressive disease (PD). Compared to the high-risk group, the low-risk group had a better overall response rate (ORR). The above experimental results suggest that CO V+TIL The risk scoring model can be used as an indicator for evaluating prognosis.

[0027] Example 2 This embodiment adds the following fourth step to the first embodiment, while the rest of the operations are the same as in the first embodiment: Step four: Based on the patient's angiogenesis-related factor and tumor-infiltrating lymphocyte co-expression score and TNM stage, the patient's risk score is obtained using the following method: (1) Patients with a co-expression score of angiogenesis-related factors and tumor-infiltrating lymphocytes of less than 28 were classified as low-level group, and those with a score of greater than 28 were classified as high-level group. The low-level group was assigned a score of 0 and the high-level group was assigned a score of 1. (2) Patients in TNM stage 1-3 are given 0 points, and those in TNM stage 4 are given 1 point; (3) Add the scores from step (1) and step (2) together to obtain the patient’s risk score.

[0028] Through step four above, the present invention, namely the construction of angiogenesis-related markers and tumor-infiltrating lymphocyte markers (CO), is achieved. V+TIL The score can be combined with TNM staging for prognostic prediction of nasopharyngeal carcinoma. A comparison between this score and TNM staging alone is as follows: Using multivariate Cox regression analysis, a CO2 regression model was established. V+TIL A prognostic scoring model that combines the score with TNM staging. CO V+TIL The low-scoring group is assigned 0 points, and the high-scoring group is assigned 1 point. Patients in TNM stages 1-3 are assigned 0 points, and those in TNM stage 4 are assigned 1 point. These two assigned scores are added together to generate a risk score. The cumulative risk score for each patient is calculated, and receiver operating characteristic (ROC) curve analysis is used to compare the prognostic scoring model with TNM staging or CO. V+TIL The sensitivity and specificity of the score model alone. TNM staging combined with CO V+TIL Comparison of score and TNM stage: OS (AUC: 0.837, 95%CI: 0.784–0.882 vs. AUC: 0.794, 95%CI: 0.737–0.844; p=0.0578); PFS (AUC: 0.840, 95%CI: 0.787–0.884 vs. AUC: 0.772; 95%CI: 0.713–0.823; p=0.0018), LRRFS (AUC: 0.706, 95%CI: 0.644–0.763 vs. AUC: 0.636, 95%CI: 0.573–0.699; p=0.061) and DMFS (AUC: 0.800, 95%CI: 0.744–0.849 vs. TNM stage). The prognostic value of AUC: 0.738, 95% CI: 0.678–0.793; p=0.073 was significantly better than that of TNM staging alone. Figure 19-21 As shown in the figure, the present invention constructs a prognostic prediction model for nasopharyngeal carcinoma by combining the detection of angiogenesis and tumor-infiltrating lymphocyte-related markers, and its prognostic value is higher than that of the prior art.

[0029] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A method for constructing a prognostic model for nasopharyngeal carcinoma by jointly detecting tumor-related biomarkers, characterized in that, The steps include the following: Step 1: Obtain a tissue section of the patient's nasopharyngeal carcinoma. Step two involves obtaining nasopharyngeal carcinoma tissue sections, which can represent CD4 markers of tumor-associated infiltrating lymphocytes. + T, CD8 + T and a group of biomarkers that can represent angiogenesis-related markers, including VEGF-A and VEGFR-2, and the biomarker CD4 were calculated. + T, CD8 + Immunohistochemical scores of T, VEGF-A, and VEGFR-2; Step 3: Calculate the co-expression score of angiogenesis-related factors and tumor-infiltrating lymphocytes based on the immunohistochemical score.

2. The method for constructing a nasopharyngeal carcinoma prognostic model by jointly detecting tumor-related biomarkers according to claim 1, characterized in that, In step two, the marker CD4 is calculated using the following method. + T, CD8 + Immunohistochemical scores of T, VEGF-A, and VEGFR-2: (1) Marker CD4 + T, CD8 + T, VEGF-A, and VEGFR-2 positive cells were scored as negative, weakly positive, moderately positive, and strongly positive, respectively, with scores of 0, 1, 2, and 3. The marker CD4 was also scored. + T, CD8 + The percentages of T, VEGF-A, and VEGFR-2 positive cells are less than 25%, between 25% and 50%, between 51% and 75%, and greater than 75%, respectively, and are scored as 1, 2, 3, and 4 points, respectively. (2) Marker CD4 + T, CD8 + The immunohistochemical score is calculated by multiplying the staining intensity scores of T, VEGF-A, and VEGFR-2 positive cells by the percentage of positive cells.

3. The method for constructing a nasopharyngeal carcinoma prognostic model by jointly detecting tumor-related biomarkers according to claim 1, characterized in that, In step three, the co-expression score of angiogenesis-related factors and tumor-infiltrating lymphocytes in the patient is calculated using the following method: CO V+TIL score = H-score VEGFA + H-score VEGFR2 +[12-(H-score CD4 )] + [(12- H-score CD8 )]; Among them, CO V+TIL The score represents the co-expression score of angiogenesis-related factors and tumor-infiltrating lymphocytes, H-score. VEGFA The H-score is used to assess the immunohistochemical score of the biomarker VEGF-A. VEGFR2 The immunohistochemical score for the biomarker VEGFR-2, H-score CD4 For the marker CD4 + T immunohistochemical score, H-score CD8 For the marker CD8 + T's immunohistochemical score.

4. The method for constructing a nasopharyngeal carcinoma prognostic model by jointly detecting tumor-related biomarkers according to any one of claims 1 to 3, characterized in that, It also includes step four: obtaining the patient's risk score based on the patient's angiogenesis-related factor and tumor-infiltrating lymphocyte co-expression score and TNM stage.

5. The method for constructing a nasopharyngeal carcinoma prognostic model by jointly detecting tumor-related biomarkers according to claim 4, characterized in that, Based on the patients' angiogenesis-related factor and tumor-infiltrating lymphocyte co-expression scores and TNM staging, the patients' risk scores were obtained using the following method: patients with angiogenesis-related factor and tumor-infiltrating lymphocyte co-expression scores less than 28 were assigned to the low group, and those with scores greater than 28 were assigned to the high group. The low group was assigned 0 points and the high group was assigned 1 point. (2) Patients in TNM stage 1-3 are given 0 points, and those in TNM stage 4 are given 1 point; (3) Add the scores from step (1) and step (2) together to obtain the patient’s risk score.

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