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Objective:To investigate the distribution of M2 tumor-associated macrophage (TAM) in hepatocellular carcinoma (HCC) and their correlation with clinicopathological features, and the significance of programmed death-ligand 1 (PD-L1) expression.Methods:From January 1, 2012 to December 31, 2020, a total of 320 HCC patients who underwent surgical resection at the Third People′s Hospital of Nantong were included. The distribution of CD163 labeled and PD-L1 CD163 double-labeled M2 TAM in HCC tissues was detected by immunohistochemistry, and the cell density was calculated. The cell density> the average cell density (112/mm 2) was judged as high-density, the cell density≤ the average cell density was judged as low-density. The correlation between CD163 positive and PD-L1 CD163 double positive M2 TAM density and the clinical pathological characteristics of HCC and its impact on prognosis were analyzed. Chi-square test was used to analyze the correlation between M2 TAM expression and the clinical pathological characteristics of HCC. Kaplan-Meier method was used to draw survival curves, and log-rank test was used for inter group comparison. Univariate and multivariate Cox proportional hazards regression analysis was used to indentify the relevant factors affecting the prognosis of HCC. Results:TAM were mainly distributed in the tumor edge stroma and tumor sinusoids, CD163 positive M2 TAM were the main macrophage subtype. PD-L1 expression was observed in CD163 positive M2 TAM in HCC tissues, and PD-L1 positive M2 TAM were mainly distributed in the tumor edge stroma. The rate of high-density CD163 positive M2 TAM in HCC tissues was 44.4% (142/320). High-density CD163 positive M2 TAM was correlated with histological grade, TNM stage, and PD-L1 expression on tumor infiltrating immune cells in HCC tissues ( χ2=4.65, 6.72 and 42.19, P=0.031, =0.011 and <0.001). High-density PD-L1 and CD163 double positive M2 TAM in HCC tissues was correlated with microvascular invasion and TNM stage ( χ2=11.96 and 8.74, P=0.001 and 0.004). The median disease-free survival (DFS) time and overall survival (OS) time of patients with high-density CD163 positive M2 TAM were 21 and 36 months, respectively, which were lower than those of patients with low-density CD163 positive M2 TAM (50 and 103 months, respectively); the median DFS time and OS time of patients with high-density PD-L1 CD163 double-positive M2 TAM were 12 and 15 months, respectively, which were lower than those of patients with low-density PD-L1 CD163 double-positive M2 TAM (28 and 45 months, respectively), and the differences were statistically significant (all log-rank tests, all P<0.001). The results of multivariate Cox proportional hazards regression analysis showed that high-density CD163 positive M2 TAM, microvascular invasion and high TNM stage were independent risk factors for evaluating DFS and OS of patients with HCC (DFS time: HR=2.408 (95% confidence interval (95% CI) 1.778 to 3.261), 2.603 (95% CI 1.860 to 3.641), 4.032 (95% CI 2.833 to 5.747), all P<0.001. OS time: HR=2.007 (95% CI 1.457 to 2.764), 4.144 (95% CI 2.881 to 5.960), 4.292 (95% CI 2.915 to 6.329), all P<0.001). Conclusions:High-density of CD163 positive M2 TAM in HCC tissues indicates high malignancy and poor prognosis, and it is an independent prognostic risk factor. The expression of PD-L1 in M2 TAM suggests stronger tumor aggressiveness and worse prognosis in HCC.
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Objective:To explore the expression of programmed death receptor ligand 2 (PD-L2) in hepatocellular carcinoma (HCC) and its relationship with clinicopathological features and prognosis of patients.Methods:The data of 344 patients with HCC who underwent surgery in the Third People's Hospital of Nantong from January 2008 to December 2016 were retrospectively analyzed. Taking HCC tissue samples to make the tissue microarray, and the expression of PD-L2 protein was detected by immunohistochemical method. The relationship between PD-L2 protein expression and clinicopathological features was analyzed. Kaplan-Meier method was used to analyze the overall survival (OS) and disease-free survival (DFS) of patients, and the prognostic factors were analyzed by univariate and multivariate Cox proportional hazards model.Results:The positive expression rate of PD-L2 protein in 344 patients with HCC was 54.4% (187/344). The positive expression of PD-L2 protein was correlated with maximum tumor diameter >3 cm ( χ2 = 8.20, P < 0.01) and high histological grade ( χ2 = 9.46, P < 0.05); OS and DFS in PD-L2 positive expression group were worse than those in PD-L2 negative expression group (OS: P = 0.001; DFS: P = 0.015). PD-L2 positive expression was not an independent adverse influencing factor for OS and DFS (OS: HR = 1.321, 95% CI 0.955-1.829, P = 0.093; DFS: HR = 1.209, 95% CI 0.990-1.624, P = 0.209). Conclusions:PD-L2 is highly expressed in HCC tissues, which may be related to the degree of malignancy. PD-L2 is not an independent risk factor for the prognosis of HCC.
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Objective:To investigate the correlation between the expression of programmed death ligand-1 (PD-L1) and clinicopathological parameters and prognosis in hepatocellular carcinoma (HCC) tissues.Methods:From January 2008 to December 2016, 344 HCC patients underwent surgery in Nantong Third Hospital Affiliated to Nantong University were enrolled. The expression of PD-L1 in paraffin HCC tissues was detected by tissue microarray and immunohistochemistry. The correlation between the expression of PD-L1 in tumor cells, tumor-infiltrating immune cells and the clinicopathological parameters and prognosis of HCC patients were analyzed. And the related factors affecting the prognosis of patients were explored. Chi-square test, log-rank test and univariate and multivariate Cox regression analysis were used for statistical analysis.Results:Positive PD-L1 located in the membrane and/or cytoplasm of HCC tumor cells and tumor-infiltrating immune cells. The positive rate of PD-L1 expression in tumor cells was 21.8%(75/344). The expression of PD-L1 in tumor cells was related to histological grade and microvascular invasion, the positive rates of PD-L1 expression of patients with histological gradeⅠ, Ⅱ and Ⅲ were 7.7% (2/26), 16.5% (19/115) and 26.6% (54/203), respectively, the positive rates of PD-L1 expression of patients with or without microvascular invasion were 29.3% (34/116) and 18.0% (41/228), respectively, and the differences were statistically significant ( χ2=7.659 and 5.787, P=0.022 and 0.016). The positive expression rate of PD-L1 in tumor-infiltrating immune cells was 47.1% (162/344). The expression of PD-L1 in tumor-infiltrating immune cells was related with microvascular invasion, the positive rates of PD-L1 expression in patients with or without microvascular invasion were 56.9% (66/116) and 42.1% (96/228), respectively, and the differences were statistically significant ( χ2=6.751, P=0.009). The median survival time of patients with negative expression of PD-L1 in HCC tumor cells was 61 months (30 months, 92 months), while that of patients with positive PD-L1 expression was 16 months(6 months, 44 months), and the difference was statistically significant ( χ2=55.722, P<0.01). The expression of PD-L1 in HCC tumor cells was an independent risk factor affecting overall survival time (hazard ratio=3.090, P<0.01). Conclusions:The expression of PD-L1 in HCC tumor cells may be related to the malignancy and invasion of HCC, and may be a potential risk factor for prognosis.
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Background and purpose: Abnormal expression of arginase 2 (ARG2) in a variety of human malignant tumors was detected. Previous studies found that ARG2 significantly increased in hepatocellular carcinoma (HCC) and was related to histological grading of HCC. This study aimed to analyze the association of ARG2 expression with cell proliferation, apoptosis and prognosis in HCC. Methods: The expression levels of ARG2 mRNA in 14 samples of HCC, paracancerous liver tissues and 14 samples of normal liver were detected by reverse transcriptionpolymerase chain reaction (RT-PCR). Tissue sections from 158 HCC patients were examined immunohistochemically for protein expression of ARG2, proliferation-related proteins (Ki-67 and cyclin D1) and apoptosis-related proteins (activated caspase-3, caspase-8 and caspase-9). Immunofluorescence double labeling method was used to detect the coexpression of ARG2 and activated caspase-3, and the colocalization between ARG2 and apoptotic cells. Patients were followed up by telephone. Results: TThe expression of ARG2 mRNA was significantly increased in HCC compared with the paracancerous liver tissues and normal liver tissues (F=27.10, P＜0.01). The expression of ARG2 was positively correlated with the expression of Ki-67 and cyclin D1, respectively (r=0.247 8, P＜0.01; r=0.372 7, P＜0.01). The expression of ARG2 was positively correlated with the expression of activated caspase-3 and caspase-8, respectively (r=0.191 0, P＜0.05; r=0.180 5, P＜0.05), but not with the caspase-9 (r=0.108 9, P＞0.05). Immunofluorescence double labeling showed that ARG2 was coexpressed with the activated caspase-3 and colocalized with apoptotic cells. Kaplan-Meier survival curves showed that the median survival time was 32 months in ARG2(-) group, 18 months in ARG2(+) group and 15 months in ARG2(++) group. The log-rank test results showed that there were significant differences in median survival time between the groups, and the median survival time in ARG2(-) group was longer than that in ARG2(+) and ARG2(++) groups (χ2=12.278, P＜0.01). Conclusion: ARG2 may be involved in regulating the proliferation and apoptosis of HCC cancer cells. Detecting the expression of ARG2 in HCC tissues may indicate prognosis.