The Role Of Beta-adrenergic In Hepatocellular Carcinoma Recurrence And Metastasis

Backgrounds:Liver cancer is one of the most malignance with high incidence and mortality.In China,liver cancer ranks first in male under 60 years old,and the overall 5-year survival rate is less than 10%.Hepatocellular carcinoma(HCC)accounts for the majority of liver cancer,which has a high recurrence and metastasis rate and lacks an effective treatment.A large amount of pre-existing clinical and animal researches have indicated that beta-blockers played an important role in tumor progression and recurrence,and it may become a potential new approach for cancer therapy.However,the effect of beta-blockers on HCC recurrence or metastasis and the mechanism of beta2 adrenergic receptor(ADRB2)in it are not fully elucidated.Aims:To study the role of beta-blockers in HCC recurrence or metastasis and to explore the possible mechanisms of beta2 adrenergic receptor(ADRB2)in it.Methods:1.100 HCC patients undergoing hepatectomy in the First Affiliated Hospital,School of Medicine,Zhejiang University(no TACE or RAF before surgery)were enrolled and divided into two groups.We analyzed and compared the prognosis of beta-blocker group(n=25)and non-beta-blocker group(n=75).2.Liver and lung metastasis mouse models were established to study the effect of ADRB2 inhibitor ICI 118,551 on HCC metastasis.3.We chose Hep-SK-1 and LM3 cell lines for in-vitro experiments,and used related lentivirus to knock down ADRB2 expression in the cells.Cell counting kit-8(CCK8)assay,wound healing assay and cell invasion assay were used to evaluate change of cell functions.We further applied mRNA sequence in shADRB2 cell line and explored the downstream mechanism of the pathway.Results:1.The analysis of correlation of ?-blockers and clinic pathological parameters showed two groups of patients were comparable.Univariate and multivariate analysis indicated gender(male)(p=0.003)and beta-blocker(p=0.043)were independent risk factors of three-year survival,while TNM stage(?+?)(p=0.008)and HBV DNA(+)(p=0.004)were independent risk factors of recurrence-free survival in this study.We also found that HCC patients treated with beta-blocker had higher recurrence-free survival rate(60.0%vs.36.0%,p=0.02)and 3-year survival rate(80.0%vs.57.3%,p=0.009).2.Animal experiment showed that ADRB2 inhibitor(ICI 118,551)groups(n=5)had less metastasis than control groups(n=5).3.In vitro experiments found that knock down ADRB2 or incubated with ICI 118,551 in Hep-SK-1 and LM3 cell lines could decrease cell proliferation and invasion ability.We used KO and KEGG analysis to further analyze the result of mRNA sequence in shADRB2 cell line and found the genes in MAPK(Erk)pathway down-regulated significantly,revealing MAPK(Erk)pathway might activate in downstream of beta-adrenergic signaling pathway.Conclusion:1.Beta-blocker could improve recurrence-free survival rate and 3-year survival rate of HCC patients while ADRB2 inhibitor could reduce HCC metastasis in mouse model,indicating beta-blocker may serve as an adjuvant therapy to HCC.2.Knock down ADRB2 reduces cell proliferation and invasion ability,of which the mechanism may through activating downstream MAPK(Erk)pathway.