β-defensin 1 expression in HCV infected liver/liver cancer: an important role in protecting HCV progression and liver cancer development

Abstract

β-defensin family plays a role in host defense against viral infection, however its role in HCV infection is still unknown. In this study, we demonstrated that β-defensin 1 was significantly reduced in HCV-infected liver specimens. Treatment with interferon and ribavirin upregulated β-defensin-1, but not other β-defensin tested, with the extent and duration of upregulation associated with treatment response. We investigated β-defensin family expression in liver cancer in publicly available datasets and found that among all the β-defensins tested, only β-defensin 1 was significantly downregulated, suggesting β-defensin 1 plays a crucial role in liver cancer development. Further analysis identified E-cadherin as the top positive correlated gene, while hepatocyte growth factor-regulated tyrosine kinase substrate as the top negative correlated gene. Expression of two proteoglycans were also positively correlated with that of β-defensin 1. We have also identified small molecules as potential therapeutic agents to reverse β-defensin 1-associated gene signature. Furthermore, the downregulation of β-defensin 1 and E-cadherin, and upregulation of hepatocyte growth factor-regulated tyrosine kinase substrate, were further confirmed in liver cancer and adjacent normal tissue collected from in-house Chinese liver cancer patients. Together, our results suggest β-defensin 1 plays an important role in protecting HCV progression and liver cancer development.

Figure 1

The expression of β-defensin 1 in normal and HCV infected liver. Box plots for mRNA expression of (A) β-defensin 1, (B) β-defensin 4 and (C) β-defensin 126 in normal and HCV-infected liver.

Figure 2

The expression of β-defensin 1 in peripheral blood mononuclear cells in patients treated with interferon and ribavirin. Error plots for mRNA expression of β-defensin 1 in peripheral blood mononuclear cells collected at different time point pre- or post-treatment with interferon and ribavirin in (A) all HCV infected patients, (B) HCV infected patients with poor response to treatment and (C) HCV infected patients with intermediate/mark response to treatment.

Figure 3

The expression of β-defensin 1 in non-tumor and tumor liver specimens. Box plots for mRNA expression of β-defensin 1 in non-tumor and tumor liver specimens in (A) GSE25097 (n = 511), (B) GSE14520 (n = 445) and (C) GSE36376 (n = 433).

Figure 4

The correlation of mRNA expression levels of β-defensin 1 and CDH1, and β-defensin 1 and HGS. Scatter plots for mRNA expression levels of DEFB1 and CDH1 in liver specimens in (A) GSE25097 (n = 557), (B) GSE14520 (n = 445) and (C) GSE36376 (n = 459). Scatter plots for mRNA expression levels of DEFB1 and HGS in liver specimens in (D) GSE25097 (n = 557), (E) GSE14520 (n = 445) and (F) GSE36376 (n = 459).

Figure 5

The correlation of mRNA expression levels of β-defensin 1 and DCN, and β-defensin 1 and LUM. Scatter plots for mRNA expression levels of DEFB1 and DCN in liver specimens in (A) GSE25097 (n = 557), (B) GSE14520 (n = 445) and (C) GSE36376 (n = 459). Scatter plots for mRNA expression levels of DEFB1 and LUM in liver specimens in (D) GSE25097 (n = 557), (E) GSE14520 (n = 445) and (F) GSE36376 (n = 459).

Figure 6

The expression of CDH1 and HGS in non-tumor and tumor liver specimens. Box plots for mRNA expression of CDH1 in non-tumor and tumor liver specimens in (A) GSE25097 (n = 511), (B) GSE14520 (n = 445) and (C) GSE36376 (n = 433). Box plots for mRNA expression of HGS in non-tumor and tumor liver specimens in (D) GSE25097 (n = 511), (E) GSE14520 (n = 445) and (F) GSE36376 (n = 433).

Figure 7

The expression of DCN and LUM in non-tumor and tumor liver specimens. Box plots for mRNA expression of DCN in non-tumor and tumor liver specimens in (A) GSE25097 (n = 511), (B) GSE14520 (n = 445) and (C) GSE36376 (n = 433). Box plots for mRNA expression of LUM in non-tumor and tumor liver specimens in (D) GSE25097 (n = 511), (E) GSE14520 (n = 445) and (F) GSE36376 (n = 433).

Figure 8

The results of connectivity mapping with the DEFB1 gene signature. (A) The raw connection scores (cscore). (B) The normalized connection scores (zscore). Each data point on these graphs represents a compound screened. The blue line indicates the threshold of statistical significance. Those data points above the blue line have significant connectivity mapping scores with the DEFB1 gene signature (significance mark = 1). Show in green are the significant drugs (listed in Supplementary Table S2) that are also highly stable under gene signature perturbations (perturbation stability = 1).

Figure 9

The results of immunohistochemical staining. (A) Representative images of immunohistochemical staining of β-defensin 1, E-cadherin and HGS in both cancerous and normal liver specimens. (B–D) The percentage of cases stained with different levels of (B) β-defensin 1, (C) E-cadherin and (D) HGS.