Tumor Biol. DOI 10.1007/s13277-014-2927-4
RESEARCH ARTICLE
Tumor-infiltrating neutrophils predict poor outcome in adenocarcinoma of the esophagogastric junction Pingping Hu & Zhaofei Pang & Hongchang Shen & Guanghui Wang & Haifeng Sun & Jiajun Du
Received: 4 October 2014 / Accepted: 28 November 2014 # International Society of Oncology and BioMarkers (ISOBM) 2014
Abstract Tumor-infiltrating neutrophil (TIN) has been reported to be an independent predictor in multiple tumors, but its role in the development of adenocarcinoma of the esophagogastric junction (AEG) remains unclear. We conducted immunohistochemistry (IHC) to detect the expression of TIN in 113 consecutive patients with primary AEG. The prognostic value and its relationship with clinicopathological characteristics and Ki-67 (a proliferation-associated antigen) expression were analyzed. High level of TIN was related to poor outcome in AEG patients with a hazard ratio (HR) of 3.93 (95 % confidence interval (CI)=2.14–7.19, p65 years Sex Male Female T stage T1–T2 T3–T4 N status N− N+ Tumor differentiation Grade 1–2 Grade 3–4 Circulating NEUTs Low High Circulating WBC Low High
No.
Percent
62 51
54.9 45.1
101 12
89.4 10.6
28 85
24.8 75.2
24 89
21.2 78.8
62 51
54.9 45.1
58
51.3
55
48.7
57 56
50.4 49.6
NEUTs neutrophils, WBC white blood cell
peripheral blood was as follows: 58 patients had a low level of circulating neutrophils while 55 had a high level. Fig. 1 The expression of TIN and Ki-67 in AEG tumor tissues. a Negative expression of TIN, in which no neutrophils infiltrated in the tumor tissue. b Positive expression of TIN, in which positive staining of TIN was localized in the nucleus of neutrophils infiltrated in the tumor tissue. c Low level of Ki-67 expression in the tumor tissue, in which positive staining of Ki-67 was localized in the nucleus of the tumor tissue. d High level of Ki-67 expression in the tumor tissue
Low level of the circulating white blood cells was found in 57 patients, and a high level was found in the remainder. Seventy-two patients had died before end of followup.
Detection of TIN and Ki-67 expression in AEG tissues by IHC IHC was used to investigate TIN infiltration and Ki-67 expression in AEG tissues. Neutrophils were observed in all tumor compartments with the most obvious infiltration in the stroma. Neutrophils localized in the areas of necrosis showed higher densities than those in areas without necrosis, while the necrotic areas were excluded when evaluation to avoid the impact of necrosis was performed. There were no obvious differences in neutrophil densities between superficial and deep portion of the same tumor. The cutoff value determined by ROC curve was 7.5, so TIN of less than or equal to seven cells/HPF was defined as low expression and TIN of greater than seven cells/HPF was defined as high expression. High level of TIN was detected in 74/113 (65.5 %) AEG patients, and high expression of Ki-67 was found in 46/113 (40.7 %) patients. The positive staining of TIN was mainly localized in the cytoplasm and plasma membrane of neutrophils infiltrating in tumor tissue, whereas that of Ki-67 was in the nucleus of tumor cells (Fig. 1).
Tumor Biol.
Survival analysis
Relationship between TIN and clinicopathological characteristics The relationship between TIN and clinicopathological parameters of 113 AEG patients was demonstrated in Table 2. High level of TIN was found to be associated with T stage (p=0.006), positive lymph node metastasis (N status, p=0.03), and high expression of Ki-67 (p= 0.008). In tumors with high TIN infiltration, lymph node metastases were observed in 63/74 tumors, while in tumors with low density, the metastases were observed in 26/39 tumors. The odds ratio (OR) of lymph node metastases was 2.86 (95 % confidence interval (CI)=1.14–7.21). On the contrary, no significant correlation was discovered between the level of TIN and other clinical parameters, such as age (p = 0.43), sex (p=0.82), grade of tumor differentiation (p=0.08), circulating neutrophils (p=0.55), and white blood cell (p= 0.11) in the peripheral blood.
Table 2
Relationship between TIN and clinicopathological data Low TIN (%)
Age ≤65 years >65 years Sex Male Female T stage T1–T2 T3–T4 N status N− N+ Tumor differentiation Grade 1–2 Grade 3–4 Ki-67 expression Low High Circulating NEUTs Low High Circulating WBC Low High
High TIN (%)
p 0.43a
19 (30.6) 20 (39.2)
43 (69.4) 31 (60.8) 0.82b
34 (33.7) 5 (41.7)
67 (66.3) 7 (58.3) 0.006a
16 (57.1) 23 (27.1)
Kaplan-Meier survival curves also demonstrated a high level of TIN expression to be a significantly unfavorable prognostic factor for overall survival (Fig. 2). According to univariate Cox regression analysis, the overall survival was correlated with TNM stage (hazard ratio (HR)=1.93, 95 % CI=1.13– 3.30, p=0.02), grade of tumor differentiation (HR=2.10, 95 % CI=1.31–3.35, p=0.002), Ki-67 expression (HR=2.15, 95 % CI=1.35–3.42, p=0.001), and TIN expression (HR=3.93, 95 % CI=2.14–7.19, p
RESEARCH ARTICLE
Tumor-infiltrating neutrophils predict poor outcome in adenocarcinoma of the esophagogastric junction Pingping Hu & Zhaofei Pang & Hongchang Shen & Guanghui Wang & Haifeng Sun & Jiajun Du
Received: 4 October 2014 / Accepted: 28 November 2014 # International Society of Oncology and BioMarkers (ISOBM) 2014
Abstract Tumor-infiltrating neutrophil (TIN) has been reported to be an independent predictor in multiple tumors, but its role in the development of adenocarcinoma of the esophagogastric junction (AEG) remains unclear. We conducted immunohistochemistry (IHC) to detect the expression of TIN in 113 consecutive patients with primary AEG. The prognostic value and its relationship with clinicopathological characteristics and Ki-67 (a proliferation-associated antigen) expression were analyzed. High level of TIN was related to poor outcome in AEG patients with a hazard ratio (HR) of 3.93 (95 % confidence interval (CI)=2.14–7.19, p65 years Sex Male Female T stage T1–T2 T3–T4 N status N− N+ Tumor differentiation Grade 1–2 Grade 3–4 Circulating NEUTs Low High Circulating WBC Low High
No.
Percent
62 51
54.9 45.1
101 12
89.4 10.6
28 85
24.8 75.2
24 89
21.2 78.8
62 51
54.9 45.1
58
51.3
55
48.7
57 56
50.4 49.6
NEUTs neutrophils, WBC white blood cell
peripheral blood was as follows: 58 patients had a low level of circulating neutrophils while 55 had a high level. Fig. 1 The expression of TIN and Ki-67 in AEG tumor tissues. a Negative expression of TIN, in which no neutrophils infiltrated in the tumor tissue. b Positive expression of TIN, in which positive staining of TIN was localized in the nucleus of neutrophils infiltrated in the tumor tissue. c Low level of Ki-67 expression in the tumor tissue, in which positive staining of Ki-67 was localized in the nucleus of the tumor tissue. d High level of Ki-67 expression in the tumor tissue
Low level of the circulating white blood cells was found in 57 patients, and a high level was found in the remainder. Seventy-two patients had died before end of followup.
Detection of TIN and Ki-67 expression in AEG tissues by IHC IHC was used to investigate TIN infiltration and Ki-67 expression in AEG tissues. Neutrophils were observed in all tumor compartments with the most obvious infiltration in the stroma. Neutrophils localized in the areas of necrosis showed higher densities than those in areas without necrosis, while the necrotic areas were excluded when evaluation to avoid the impact of necrosis was performed. There were no obvious differences in neutrophil densities between superficial and deep portion of the same tumor. The cutoff value determined by ROC curve was 7.5, so TIN of less than or equal to seven cells/HPF was defined as low expression and TIN of greater than seven cells/HPF was defined as high expression. High level of TIN was detected in 74/113 (65.5 %) AEG patients, and high expression of Ki-67 was found in 46/113 (40.7 %) patients. The positive staining of TIN was mainly localized in the cytoplasm and plasma membrane of neutrophils infiltrating in tumor tissue, whereas that of Ki-67 was in the nucleus of tumor cells (Fig. 1).
Tumor Biol.
Survival analysis
Relationship between TIN and clinicopathological characteristics The relationship between TIN and clinicopathological parameters of 113 AEG patients was demonstrated in Table 2. High level of TIN was found to be associated with T stage (p=0.006), positive lymph node metastasis (N status, p=0.03), and high expression of Ki-67 (p= 0.008). In tumors with high TIN infiltration, lymph node metastases were observed in 63/74 tumors, while in tumors with low density, the metastases were observed in 26/39 tumors. The odds ratio (OR) of lymph node metastases was 2.86 (95 % confidence interval (CI)=1.14–7.21). On the contrary, no significant correlation was discovered between the level of TIN and other clinical parameters, such as age (p = 0.43), sex (p=0.82), grade of tumor differentiation (p=0.08), circulating neutrophils (p=0.55), and white blood cell (p= 0.11) in the peripheral blood.
Table 2
Relationship between TIN and clinicopathological data Low TIN (%)
Age ≤65 years >65 years Sex Male Female T stage T1–T2 T3–T4 N status N− N+ Tumor differentiation Grade 1–2 Grade 3–4 Ki-67 expression Low High Circulating NEUTs Low High Circulating WBC Low High
High TIN (%)
p 0.43a
19 (30.6) 20 (39.2)
43 (69.4) 31 (60.8) 0.82b
34 (33.7) 5 (41.7)
67 (66.3) 7 (58.3) 0.006a
16 (57.1) 23 (27.1)
Kaplan-Meier survival curves also demonstrated a high level of TIN expression to be a significantly unfavorable prognostic factor for overall survival (Fig. 2). According to univariate Cox regression analysis, the overall survival was correlated with TNM stage (hazard ratio (HR)=1.93, 95 % CI=1.13– 3.30, p=0.02), grade of tumor differentiation (HR=2.10, 95 % CI=1.31–3.35, p=0.002), Ki-67 expression (HR=2.15, 95 % CI=1.35–3.42, p=0.001), and TIN expression (HR=3.93, 95 % CI=2.14–7.19, p
