Prognostic significance of synergistic hexokinase-2 and beta2-adrenergic receptor expression in human hepatocelluar carcinoma after curative resection

Abstract

Background: Hexokinase-2 (HK2) and Beta2-adrenergic receptor (Beta2AR) are overexpressed in hepatocellular carcinoma (HCC) tissues and associated with poor prognosis. However, the synergistic effect of HK2 and Beta2AR in HCC prognosis is not elucidated. The present study aims to investigate the association between HK2 and Beta2AR expressions in HCC tissues, and to evaluate the synergistic effect of HK2 and Beta2AR in HCC prognosis.

Methods: Immunohistochemistry of HK2 and Beta2AR was performed on 155 paraffin embedded HCC samples retrieved from the archives of pathology department. Corresponding clinical data and prognostic data were collected through searching medical record systems, death registration systems and interviews with patient families. Spearman correlation test was performed to evaluate the association between HK2 and Beta2AR expression. Kaplan-Meier survival curves and Cox regressions were employed to evaluate HK2 and Beta2AR expression in HCC prognosis, respectively and synergistically.

Results: 109 of 155 HCC patients reached the death point, the survival time of HCC patients was 46.23 ± 31.01 months after curative surgical resections of HCC. Kaplan-Meier survival analysis showed that large tumor size (more than 5 cm) (hazard ratio (HR) = 8.42, 95 % confidence interval (CI) = 3.81-18.59, P < 0.0001), advanced TNM stage (III and IV stages) (HR = 2.09, 95%CI = 1.21-3.62, P < 0.001) and AFP more than 20 μg/L (HR = 1.49, 95%CI = 1.02-2.18, P = 0.0302) were predictors for poor prognosis. HK2 and Beta2AR positive expression was detected in 66 (42.58) and 122 (78.71 %) HCC samples respectively. In univariate analysis, HK2(+) (HR = 2.70, 95%CI = 1.76-4.15, P < 0.0001) and Beta2AR(+) (HR = 4.61, 96%CI = 3.14-6.76, P < 0.0001) were associated with poor prognosis. In multivariate analysis, HK2(+) (P < 0.0001) and Beta2AR(+) (P < 0.0001) were also associated with poor prognosis. HK2(+)/Beta2AR(+) in HCC samples had poorer prognosis compared with HK2(-)/Beta2AR(-) in both univariate analysis (HR = 4.69, 95%CI = 2.91-7.57, P < 0.0001) and multivariate analysis (P < 0.0001). HK2(+)/Beta2AR(+) in HCC samples had poorer prognosis compared with HK2(-)/Beta2AR(+) in both univariate analysis (HR = 1.76, 95%CI = 1.17-2.64, P = 0.003) and multivariate analysis (P = 0.004).

Conclusion: HK2 and Beta2AR play important roles in HCC progression. HK2 and Beta2AR expression in HCC is correlated positively. Beta2AR may increase HCC invasion and metastasis in collaboration with HK2. HK2 and Beta2AR can predict HCC prognosis both independently and synergistically.

Keywords: Beta2-adrenergic receptor; Hepatocellular carcinoma; Hexokinase-2; Immunohistochemistry; Prognosis.

Fig. 1

Immunochemistry of HCC tissues. a Representative positive staining of HK2 in HCC tissues. b Representative negative staining of HK2 in HCC tissues. c Representative positive staining of Beta2AR in HCC tissues. d Representative negative staining of Beta2AR in HCC tissues

Fig. 2

Spearman correlation between HK2 and Beta2AR. Spearman correlation test shows that HK2 and Beta2AR expression is correlated positively (P < 0.0001)

Fig. 3

Survival curve. a and b HK2(+) is associated with poor prognosis in both univariate analysis (HR = 2.70, 95%CI = 1.76–4.15, P < 0.0001) and multivariate analysis (P < 0.0001). c and d Beta2AR(+) is associated with poor prognosis in both univariate analysis (HR = 4.61, 96%CI = 3.14–6.76, P < 0.0001) and multivariate analysis (P < 0.0001). e and f HK2(+)/Beta2AR(+) in HCC samples shows poorer prognosis as compared with HK2(−)/Beta2AR(−) in both univariate analysis (HR = 4.69, 95%CI = 2.91–7.57, P < 0.0001) and multivariate analysis (P < 0.0001). HK2(+)/Beta2AR(+) in HCC samples shows poorer prognosis as compared with HK2(−)/Beta2AR(+) in both univariate analysis (HR = 1.76, 95%CI = 1.17–2.64, P = 0.003) and multivariate analysis (P = 0.004)

Fig. 4

The difference of live time among different groups. One-way ANOVA analysis shows that survival time of HK2(−)/Beta2AR(−) is longer than that of HK2(−)/Beta2AR(+) and HK2(+)/Beta2AR(+) significantly. The survival time of HK2(−)/Beta2AR(+) is also longer than that of HK2(+)/Beta2AR(+). Data are shown with mean ± SD. ## P < 0.0001 vs. HK2(+)/Beta2(+) group, ** P < 0.0001 vs. HK2(−)/Beta2(+) group, #P = 0.001 vs. HK2(+)/Beta2AR(+) group