KR102043468B1 - An Information Providing Method for Predicting Recurrence of Epstein-Barr virus-associated Gastric Cancer - Google Patents

Abstract

The present invention relates to a method for providing information for predicting the recurrence probability of EBV-associated gastric cancer. According to the information providing method, personalized medicine that accurately predicts the recurrence potential of patients with EBV-associated gastric cancer can be managed more effectively. This can be useful for providing.

Description

An Information Providing Method for Predicting Recurrence of Epstein-Barr virus-associated Gastric Cancer}

The present invention relates to an information providing method for predicting the recurrence probability of EBV associated gastric cancer.

Epstein-Barr virus (EBV) is a human gamma-1 herpesvirus, a lifelong, endogenous opportunistic virus that infects more than 90% of the world's population. During the incubation period, the virus is mostly bound to chromatin and multicopy episomes in the nuclei of various tumor cells, such as B cells, T cells, NK cells and epithelial cells. It is reported to exist. During the incubation period can cause various malignancies, such as Burkitts lymphoma, Hodgkins disease, post-transplant lymphoproliferative disease, malignant lymphoma (NK / T) Cell lymphoma, nasopharyngeal carcinoma and gastric carcinoma have been reported.

Meanwhile, about 10% of the EBV-related diseases are diagnosed with gastric cancer. In addition, in cases of EBV associated gastric cancer, almost all gastric cancer cells are infected with EBV, and more than 50,000 cases of gastric cancer associated with EBV occur worldwide each year. Thus, gastric cancer associated with EBV is considered to be the most common case of tumors caused by EBV. However, the detailed clinical pathological characteristics of EBV associated gastric cancer have not been fully understood, and there are no studies on the clinical features and prognosis according to the expression of EBV associated gastric cancer and related factors.

Accordingly, the present inventors completed the present invention by identifying an information providing method for predicting the recurrence probability of EBV-associated gastric cancer.

Republic of Korea Patent Publication No. 10-2015-0020189

One object of the present invention is to prepare a tissue sample of a patient with Epstein-Barr virus (EBV) associated gastric cancer; The programmed cell death protein ligand-1 (PD-L1), the programmed cell death protein ligand-2 (PD-L2) and tumor-infiltrating lymphocytes in the sample identifying expression levels of one or more proteins selected from the group consisting of tumour-infiltrating lymphocytes (TILs); And it provides an information providing method for predicting the recurrence probability of EBV-associated gastric cancer comprising the step of comparing the expression level with the control.

One aspect of the present invention comprises the steps of preparing a tissue sample of Epstein-Barr virus (EBV) associated gastric cancer patient; The programmed cell death protein ligand-1 (PD-L1), the programmed cell death protein ligand-2 (PD-L2) and tumor-infiltrating lymphocytes in the sample identifying expression levels of one or more proteins selected from the group consisting of tumour-infiltrating lymphocytes (TILs); And it provides an information providing method for predicting the recurrence probability of EBV-associated gastric cancer comprising the step of comparing the expression level with the control.

The information providing method of the present invention can provide information on patients with high probability of relapse among patients with EBV-associated gastric cancer, especially those with EBV-associated gastric cancer who have undergone gastrectomy. This can be useful to provide.

The first step of the invention is to prepare a tissue sample of an EBV associated gastric cancer patient.

Tissue samples from EBV associated gastric cancer patients can be prepared using methods commonly used in the art.

According to an embodiment of the present invention, the gastric cancer patient may be a patient who has undergone gastrectomy.

According to one embodiment of the invention, the resection may be surgical resection or endoscopic submucosal dissection (ESD).

An EBV-associated gastric cancer patient for informational purposes may be a patient who has undergone surgical surgery, radiation therapy and / or chemotherapy, but is preferably a patient who has undergone surgical surgery, particularly gastrectomy. In particular, patients who have undergone surgical resection and / or endoscopic mucosal resection during gastrectomy can provide more accurate information about the prognosis of EBV-associated gastric cancer.

The second step of the present invention is programmed cell death protein ligand-1 (PD-L1), programmed cell death protein ligand-2 (PD) in the sample -L2) and tumor-infiltrating lymphocyte (TIL) to identify the expression level of one or more proteins selected from the group consisting of.

According to one embodiment of the invention, the confirmation may be by immunohistochemistry.

To provide information about the likelihood of recurrence in patients with EBV associated gastric cancer, expression levels of PD-L1, PD-L2 and TIL proteins can be measured from tissue samples of EBV associated gastric cancer patients prepared in the first step. For example, western blotting, immunocytochemistry, radioimmunoassay (RIA), radioimmunodiffusion, enzyme immunoassay (ELISA), immunoprecipitation, Industry such as flow cytometry, immunofluorescence, ouchterlony, complement fixation assay, protein chip and / or immunohistochemistry The expression level of the protein may be measured using a general protein quantification method, but preferably, immunohistochemistry may be used, but is not limited thereto.

On the other hand, the expression level of PD-L1, PD-L2 and TIL protein can be measured by the method of measuring the mRNA expression level of each protein. For example, polymerase chain reaction (PCR), reverse transcriptase polymerase chain reaction (RT-PCR), real-time polymerase chain reaction (Real-time PCR), RNase protection assay (RPA), microarray ) And / or Northern blotting can be used to determine the expression level of the protein using methods that measure the general mRNA expression level in the art.

The third step of the present invention is to compare the expression level with the control.

The comparison of this step may be compared with the expression level of the positive control group or the negative control group, and when using a plurality of EBV patient samples, a method of comparing the expression level of each sample relative to the expression level of the entire sample group It can be substituted for comparison with the control.

According to one embodiment of the present invention, after the comparing step, when the expression of PD-L1 is increased compared to the control, it is determined that the likelihood of recurrence of EBV-associated gastric cancer is high, and when compared with the control group of EBV-associated gastric cancer The method may further include determining that the probability of recurrence is low.

According to one embodiment of the present invention, after the comparing step, when the expression of the PD-L2 is reduced compared to the control, it is determined that the likelihood of recurrence of EBV-associated gastric cancer is high, and when compared with the control group of EBV-associated gastric cancer The method may further include determining that the probability of recurrence is low.

According to one embodiment of the present invention, after the comparing step, when the expression of the TIL is reduced compared to the control, it is determined that the likelihood of recurrence of EBV-associated gastric cancer is high, and when it is increased compared to the control, the possibility of recurrence of EBV-associated gastric cancer The method may further include determining that it is low.

The expression levels of PD-L1, PD-L2 and TIL proteins can be used to provide information on the possibility of recurrence of EBV-associated gastric cancer, respectively. It is possible.

According to one embodiment of the invention, the comparison may be made in tumor cells (intraepithelial tumourcell) in the epithelium.

According to one embodiment of the invention, the comparison can be made in the tumor stroma area.

For PD-L1 protein, the expression level in tumor cells is associated with the prognosis of EBV associated gastric cancer, and for PD-L2 and TIL proteins, the expression level in tumor stroma is associated with the prognosis of EBV associated gastric cancer. It is preferable to compare the amount expressed in tumor cells with the control group, and the amount of PD-L2 and TIL protein expressed in the tumor substrate is preferably compared with the control group.

According to the information providing method of the present invention, it is possible to accurately predict the recurrence possibility of EBV-associated gastric cancer patients, it can be usefully used to provide personalized medicine that can be more effective patient management.

1 is a photograph showing immunohistochemical expression of PD-L1 and PD-L2 in EBV-associated gastric cancer, where A to F are iTu-PD-L1 negative or str-PD-L1 negative (A) and iTu-PD-, respectively. L1 positive (B), str-PD-L1 positive (C), iTu-PD-L2 negative or str-PD-L2 negative (D), iTu-PD-L2 positive and str-PD-L2 positive.
FIG. 2 is a Kaplan-Meier survival curve showing stratified disease free survival of iTu-PD-L1 positive and iTu-PD-L1 negative patients.
FIG. 3 is a Kaplan-Meyer survival curve showing stratified disease free survival of iTu-PD-L2 positive and iTu-PD-L2 negative patients.
4 is a Kaplan-Meyer survival curve showing stratified disease-free survival of str-PD-L2 positive and str-PD-L2 negative patients.
5 is a Kaplan-Meier survival curve showing stratified disease-free survival of iTu-PD-L1 negative and str-TIL positive, iTu-PD-L1 positive and str-TIL negative and other patients.

Hereinafter, one or more embodiments will be described in more detail with reference to Examples. However, these examples are provided to illustrate one or more embodiments illustratively and the scope of the present invention is not limited to these examples.

Example  One. EBV Association of Gastric Cancer  Prognostic Evaluation Method

<1-1> Patient and Sample Preparation

A total of 120 EBV-associated gastric cancer (EBVaGC) patients were used to evaluate the prognosis of EBVaGC using known criteria for patient selection and outcome (Kang et al, 2016). .

Specifically, the Institutional Review Board (IRB, No .: KNUMC 2016-05-012) of Kyungpook National University Medical Center approved the study, and from January 2011 to November 2014, Kyungpook National University Medical Center (KNUMC) Surgical resection or endoscopic submucosal dissection (ESD) was performed and samples were collected from all patients histologically diagnosed with gastric adenocarcinoma). EBV positive status was determined through the EBV-encoded RNA in situ hybridization method. Clinical and pathological information was collected by reviewing the patient's medical records and pathological reports and in accordance with the American Joint Committee on Cancer's Staging Manual for Stomach cancer (Edge et al, 2010). Patients were staged and followed the Reporting Recommendations for Tumourmarker Prognostic Studies criteria, McShane et al, 2005.

<1-2> Immunohistochemistry and Analysis

All tissue slides were stained using an automatic immunohistochemistry (IHC) staining device (BenchMarkXT, Ventana Medical Systems, Tucson, AZ, USA) according to the manufacturer's instructions, using the primary antibodies shown in Table 1 below. It was.

division Antibody Name Dilution Ratio Remarks PD-L1 E1L3N 1: 100 Cell Signaling Technology, Danvers, MA, USA PD-L2 clone 176611 1: 150 R & D Systems, Minneapolis, MN, USA

Specifically, normal gastric cancer tissue samples and each representative tissue samples collected in Example <1-1> were formalin-fixed and paraffin-embedded, cut into 4 mm thick sections, and poly-L-lysine-coated adhesive slides. It was transferred to and then microwaved at 60 ℃ for 1 hour. The sections were then deparaffinized using EZ Prep solution (Ventana Medical Systems). For antigen recovery, after treatment with cell-conditioning solution 1 (CC1) standard reagent (pH 8.4 buffer containing Tris / borate / EDTA), inhibitor D (3% H) at 37 ° C. for 4 minutes. ₂ O ₂ ) was blocked. The slides were then reacted with the primary antibody at 37 ° C. for 60 minutes. Staining was optimized using positive and negative controls for each marker according to manufacturer's instructions. All sections were visualized with the UltraView Universal DAB Detection Kit (Ventana Medical Systems). The expression of PD-L1 and PD-L2 was assessed by two experienced gastrointestinal pathologists (ANS and HIB) without prior knowledge of clinical data and the development of tumor cells and tumor stroma areas (immune cells). To confirm the intensity and percentage of membrane and / or cytoplasmic staining, the stained slides were scored semiquantitatively and each marker was interpreted according to the criteria in Table 2 below.

division standard 0 Less than 1% or no expression of immune cells in iTu- or str- 1+ Weak or Faint Expression of Immune Cells in iTu- or str- (in Area of 1%) 2+ Moderate expression of immune cells in iTu- or str- (in 1% area) 3+ Strong expression of immune cells in iTu- or str- (at 1% area)

Each sample was then classified into two covariates, negative (0 and 1+) and positive (2+ and 3+).

<1-3> Of TILs  Pathological Quantification

Percentages of tumor-infiltratinglymphocytes (TIL) and tumor stroma (str-) of intuepithelial tumour cells (iTu-) in the TIL score table of the International Breast Cancer TILs Working Group 2014 Was evaluated using a modified version (Dieci et al, 2014), and the detailed quantitative meaning is as previously described (Kang et al, 2016).

Specifically, iTu-TILs are defined as the percentage of mononuclear inflammatory cells that are in direct contact with tumor cells within the epithelial cell nest. On the other hand, str-TILs are defined as the ratio of tumor stroma to invasive carcinoma occupied by monocyte inflammatory cells. Evaluation was made when the ratio of TIL was 10% or more, and when the percentage of iTu-TILs and str-TILs was less than 10%, 1% or 5% of the baseline for the entire section of hematoxylin and eosin stained slides. Was applied. Two pathologists (ANS and HIB), iTu-TILs and str-, of normal gastric cancer tissue and respective representative tissue samples collected in Example 1-1 above, according to conventional data (Kang et al, 2016). TILs were quantified.

<1-4> Statistical Analysis

Descriptive statistics are expressed as percentages and median values. Categorical variables were evaluated using the appropriate χ ² -test, Fisher's exact test, or Pearson's correlation (R) test. Disease-free survival (DFS) was measured from surgery or ESD to death after tumor recurrence or any cause. Overall survival (OS) was measured from postoperative or ESD to death from any cause. At the last follow-up the patient was censored or the data was censored if the patient was alive. Disease free survival and overall survival were compared through multivariate survival analysis based on Kaplan-Meier test, Log-rank test, and Cox's proportional hazard regression model. Hazard ratios (HRs) and 95% confidence intervals (CIs) were estimated for each factor. All tests were performed two-sided and considered to be statistically significant when P <0.05. All statistical analyzes were performed using the Statistical Package for Social Science for Windows (SPSS, version 19.0, SPSS Inc., Chicago, IL, USA).

Example  2. Confirmation of PD-L1 and PD-L2 Expression

In order to confirm the pattern of PD-L1 and PD-L2 expression in EBVaGC patients, the method of Example <1-2> was confirmed whether the PD-L1 and PD-L2 positive or negative in the sample of EBVaGC patients.

As a result, the immunohistochemistry of PD-L1 and PD-L2 was confirmed in 116 of 120 cases (4 cases excluded due to tissue loss or poor IHC staining). Immunohistochemical expression of PD-L1 and PD-L2 was observed in immune cells in the intepithelial (iTu-) and / or stroma area (str-) (FIG. 1). iTu-PD-L1 positive was found in 57 of 116 EBVaGC cases (49.1%, 2+: 39 cases, 3+: 18 cases), but str-PD-L1 positive was 66 cases (56.9%, 2+ 65 cases) , 3+ 1 cases). In addition, 26 EBVaGCs showing iTu-PD-L1 and str-PD-L1 were observed in 2 cases (22.4%).

On the other hand, iTu-PD-L2 positive was observed in 25 cases (21.6%, 2+: 25 cases), but str-PDL2 positive was observed in 45 cases (38.8%, 2+: 45 cases), iTu-PD- EBVaGC simultaneously showing L2 and str-PD-L2 were observed in 14 cases (12.1%).

That is, through the above results, it was confirmed that PD-L1 and PD-L2 expressed independently.

Example  3. Clinical pathology and With TIL  Association check

In order to confirm the association between TIL and the clinicopathological characteristics of EBVaGC patients, TIL was quantified in samples of EBVaGC patients with PD-L1 and PD-L2 positive or negative by the method of Example <1-3>. Association with pathological features was analyzed.

As a result, iTu-PD-L1 positive was significantly correlated with lymph node (LN) metastasis (P = 0.012) (Table 3), while str-PD-L1 positive was str-TIL positive (P = 0.003). ), iTu-TIL positive (P = 0.004) and gastric carcinoma with lymphoid stroma (GCLS) (P = 0.024), and correlated with categories related to dense TILs (Table 4).

On the other hand, str-PD-L2 positive showed low pT category (P = 0.003), no lymph node metastasis (P = 0.0032), perineural invasion (P = 0.028), str-TIL positive (P <0.001), iTu -TIL positive (P <0.001), GCLS (P <0.001) and conventional adenocarcinoma (CA, P <0.001) were correlated (Table 5), but iTu-PDL2 positive was Crohn's-like lymph response (Crohn's disease-like lymphoid reaction (CLR), lymphoma, lymphoepithelioma-like carcinoma (LELC), etc.) were not associated with any clinicopathological features (Table 6).

division iTu-PD-L1 Positive (n = 57) Voice (n = 59) P -value age <62 25 (43.9) 30 (50.8) 0.451 》 62 32 (56.1) 29 (49.2) gender male 43 (75.4) 50 (84.7) 0.209 female 14 (24.6) 9 (15.3) Lauren classification Long 9 (15.8) 15 (25.4) 0.434 Less than 42 (73.7) 39 (66.1) Mixed 6 (10.5) 5 (8.5) ordnance T1 and T2 35 (61.4) 41 (69.5) 0.360 T3 and T4 22 (38.6) 18 (30.5) Lymph node metastasis Nonexistence 35 (61.4) 49 (83.1) 0.012 existence 22 (38.6) 10 (16.9) Lymphatic invasion
Nonexistence 34 (59.6) 42 (71.2) 0.191 existence 23 (40.4) 17 (28.8) Nerve infiltration Nonexistence 37 (64.9) 38 (64.4) 0.955 existence 20 (35.1) 21 (35.6) Semen infiltration Nonexistence 55 (96.5) 55 (93.2) 0.426 existence 2 (3.5) 4 (6.8) str-TILs
(cutoff, 25%) voice 22 (38.6) 23 (39.0) 1.000 positivity 35 (61.4) 36 (61.0) iTu-TILs
(cutoff, 27.5%) voice 28 (49.1) 29 (29.2) 1.000 positivity 29 (50.9) 30 (50.8) WHO classification Non-GCLS 27 (47.4) 30 (50.8) 0.715 GCLS 30 (52.6) 29 (49.2) Histological Subclassification LELC 16 (28.1) 19 (32.2) 0.294 CLR 22 (38.6) 28 (47.5) CA 19 (33.3) 12 (20.3)

division str-PD-L1 Positive (n = 66) Voice (n = 50) P -value age <62 31 (47.0) 24 (48.0) 0.912 》 62 35 (53.0) 26 (52.0) gender male 53 (80.3) 40 (80.0) 0.968 female 13 (19.7) 10 (20.0) Lauren classification Long 11 (16.7) 13 (26.0) 0.438 Less than 49 (74.2) 32 (64.0) Mixed 6 (9.1) 5 (10.0) ordnance T1 and T2 45 (68.2) 31 (62.0) 0.488 T3 and T4 21 (31.8) 19 (38.0) Lymph node metastasis Nonexistence 50 (75.8) 34 (68.0) 0.355 existence 16 (24.2) 16 (32.0) Lymphatic invasion
Nonexistence 44 (66.7) 32 (64.0) 0.765 existence 22 (33.3) 18 (36.0) Nerve infiltration Nonexistence 44 (66.7) 31 (62.0) 0.603 existence 22 (33.3) 19 (38.0) Semen infiltration Nonexistence 63 (95.5) 47 (94.0) 0.726 existence 3 (4.5) 3 (6.0) str-TILs
(cutoff, 25%) voice 18 (27.3) 27 (54.0) 0.004 positivity 48 (72.7) 23 (46.0) iTu-TILs
(cutoff, 27.5%) voice 24 (36.4) 33 (66.0) 0.003 positivity 42 (63.6) 17 (34.0) WHO classification Non-GCLS 26 (39.4) 31 (62.0) 0.024 GCLS 40 (60.6) 19 (38.0) Histological Subclassification LELC 22 (33.3) 13 (26.0) 0.314 CLR 30 (45.5) 20 (40.0) CA 14 (21.2) 17 (34.0)

division iTu-PD-L2 Positive (n = 25) Negative (n = 91) P -value age <62 14 (56.0) 41 (45.1) 0.372 》 62 11 (44.0) 50 (54.9) gender male 17 (68.0) 76 (83.5) 0.096 female 8 (32.0) 15 (16.5) Lauren classification Long 7 (28.0) 17 (18.7) 0.237 Less than 14 (56.0) 67 (73.6) Mixed 4 (16.0) 7 (7.7) ordnance T1 and T2 20 (80.0) 56 (61.5) 0.100 T3 and T4 5 (20.0) 35 (38.5) Lymph node metastasis Nonexistence 21 (84.0) 63 (69.2) 0.207 existence 4 (16.0) 28 (30.8) Lymphatic invasion
Nonexistence 17 (68.0) 59 (64.8) 0.817 existence 8 (32.0) 32 (35.2) Nerve infiltration Nonexistence 19 (76.0) 55 (61.5) 0.239 existence 6 (24.0) 35 (38.5) Semen infiltration Nonexistence 25 (100.0) 85 (93.4) 0.338 existence 0 (0.0) 6 (6.6) str-TILs
(cutoff, 25%) voice 10 (40.0) 35 (38.5) 1.000 positivity 15 (60.0) 56 (61.5) iTu-TILs
(cutoff, 27.5%) voice 16 (64.0) 41 (45.1) 0.116 positivity 9 (36.0) 50 (54.9) WHO classification Non-GCLS 12 (48.0) 45 (49.5) 1.000 GCLS 13 (52.0) 46 (50.5) Histological Subclassification LELC 6 (24.0) 29 (31.9) 0.329 CLR 14 (56.0) 36 (39.6) CA 5 (20.0) 26 (28.6)

division str-PD-L2 Positive (n = 45) Voice (n = 71) P -value age <62 21 (46.7) 34 (47.9) 0.100 》 62 24 (53.3) 37 (52.1) gender male 34 (75.6) 59 (83.1) 0.347 female 11 (24.4) 12 (16.9) Lauren classification Long 10 (22.2) 14 (19.7) 0.365 Less than 33 (73.3) 48 (67.6) Mixed 2 (4.4) 9 (12.7) ordnance T1 and T2 37 (82.2) 39 (54.9) 0.003 T3 and T4 8 (17.8) 32 (45.1) Lymph node metastasis Nonexistence 38 (84.4) 46 (64.8) 0.032 existence 7 (15.6) 25 (35.2) Lymphatic invasion
Nonexistence 33 (73.3) 43 (60.6) 0.168 existence 12 (26.7) 28 (39.4) Nerve infiltration Nonexistence 35 (77.8) 40 (56.3) 0.028 existence 10 (22.2) 31 (43.7) Semen infiltration Nonexistence 45 (100.0) 65 (91.5) 0.080 existence 0 (0.0) 6 (8.5) str-TILs
(cutoff, 25%) voice 8 (17.8) 37 (52.1) <0.001 positivity 37 (82.2) 34 (47.9) iTu-TILs
(cutoff, 27.5%) voice 12 (26.7) 45 (63.4) <0.001 positivity 33 (73.3) 26 (36.6) WHO classification Non-GCLS 9 (20.0) 48 (67.6) <0.001 GCLS 36 (80.0) 23 (32.4) Histological Subclassification LELC 22 (48.9) 13 (18.3) <0.001 CLR 19 (42.2) 31 (43.7) CA 4 (8.9) 27 (38.0)

On the other hand, in order to determine the relationship between PD-L1 and PD-L2 expression in EBVaGC, as a result of analyzing the correlation between PD-L1 and PD-L2 expression levels (Table 7), PD-L1 (iTu-PD- ITu expression of L1 positive and str-PD-L1: R = -0.224, P = 0.016) and PD-L2 (iTu-PD-L1 positive and str-PD-L1: R = 0.185, P = 0.047) showed PD- It was confirmed that it is related to str expression of each of L1 and PD-L2.

R (P-value) iTu-PD-L1 Str-PD-L1 iTu-PD-L2 Str-PD-L2 iTu-PD-L1 - -0.224 (0.016) 0.114 (0.224) -0.146 (0.119) Str-PD-L1 -0.224 (0.016) - -0.094 (0.315) 0.086 (0.361) iTu-PD-L2 0.114 (0.224) -0.094 (0.315) - 0.185 (0.047) Str-PD-L2 -0.146 (0.119) 0.086 (0.361) 0.185 (0.047) -

Example  4. PD-L1, PD-L2 and Patient Prognosis Of TIL  Association check

In order to confirm the relationship between PD-L1, PD-L2, and TIL and the prognosis of EBVaGC patients, the patients who sampled in Example <1-1> were followed and analyzed for recurrence of EBVaGC.

As a result, at the time of the last follow-up (April 2016), the median follow-up period of survival analysis was 36.2 months (range: 6.4-66.6), and OS and DFS after 3 years were 92.2% and 88.5%, respectively. . During the analysis period, 13 patients (10.8%) relapsed and 13 patients (10.8%) died. In univariate analysis, iTu-PD-L1 positive was closely related to more severe DFS (P = 0.032), not OS (P = 0.482) (FIG. 2), Cox's proportional adjusted to existing clinical covariates. Multivariate analysis using the hazard model confirmed that iTu-PD-L1 positive was an independent poor prognostic factor of DFS (HR = 12.085, 95% CI = 2.013-72.559, P = 0.006).

On the other hand, although not statistically significant, both iTu-PD-L2 positive and str-PD-L2 positive showed favorable DFS (iTu-PD-L2: P = 0.060 and str-PDL-2: P = 0.073) (FIG. 3 and 4). When str-TILs correlated with DFS, patients were divided into three groups (iTu-PD-L1 negative, str-TIL positive and iTu-PD-L1 positive, str-TIL negative and others). In the group of patients with iTu-PD-L1 positive and str-TIL negative showed shorter DFS (P = 0.006) than the other group (FIG. 5).

In other words, it was confirmed that the prognosis was poor when iTu-PD-L1 was positive among EBVaGC patients, and especially when iTu-PD-L1 was positive and str-TIL was negative. On the other hand, it was confirmed that the prognosis was good when iTu-PD-L2 or str-PD-L2 was positive.

So far I looked at the center of the preferred embodiment for the present invention. Those skilled in the art will appreciate that the present invention can be implemented in a modified form without departing from the essential features of the present invention. Therefore, the disclosed embodiments should be considered in descriptive sense only and not for purposes of limitation. The scope of the present invention is shown not in the foregoing description but in the claims, and all differences within the scope equivalent thereto should be construed as being included in the present invention.

Claims (9)

Preparing a tissue sample of an Epstein-Barr virus (EBV) associated gastric cancer patient;
Programmed cell death protein ligand-1 (PD-L1), programmed cell death protein ligand-2 (PD-L2) and tumor-infiltrating lymphocytes in the sample confirming the expression level of the tumor-infiltrating lymphocyte (TIL) protein;
Comparing the expression level with a control; And
When the expression of PD-L1 is increased compared to the control group and the expression of PD-L2 and TIL is decreased compared to the control group, it is determined that the likelihood of recurrence of EBV-associated gastric cancer is high.
When the expression of PD-L1 is decreased compared to the control, and the expression of PD-L2 and TIL is increased compared to the control, determining that the probability of recurrence of EBV-associated gastric cancer is low
Including,
Comparison of the PD-L1 is made in tumor cells (intraepithelial tumour cells),
Comparison of the PD-L2 is made in the tumor cells or tumor stroma area in the epithelium,
The comparison of the TIL is made on the tumor stroma,
Said patient is Korean
Informational method for predicting recurrence of EBV associated gastric cancer. delete delete delete delete delete The method of claim 1, wherein the gastric cancer patient is a patient who has undergone gastrectomy. 10. The method of claim 7, wherein the resection is surgical resection or endoscopic submucosal dissection (ESD). The method of claim 1, wherein said identification is by immunohistochemistry.

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