Oncolytic Vaccinia Virus Carrying Aphrocallistes vastus Lectin (oncoVV-AVL) Enhances Inflammatory Response in Hepatocellular Carcinoma Cells

Abstract

Aphrocallistes vastus lectin (AVL) is a C-type marine lectin derived from sponges. Our previous study demonstrated that oncolytic vaccinia virus carrying AVL (oncoVV-AVL) significantly enhanced the cytotoxicity of oncoVV in cervical cancer, colorectal cancer and hepatocellular carcinoma through the activation of Ras/ERK, MAPK/ERK and PI3K/Akt signaling pathways. In this study, the inflammatory response induced by oncoVV-AVL in a hepatocellular carcinoma cell (HCC) model was investigated. The results showed that oncoVV-AVL increased the levels of inflammatory cytokines including IL-6, IL-8 and TNF-α through activating the AP-1 signaling pathway in HCC. This study provides novel insights into the utilization of lectin AVL in the field of cancer therapy.

Keywords: AP-1; Aphrocallistes vastus lectin; hepatocellular carcinoma; oncolytic vaccinia virus.

Figure 1

oncoVV-AVL increased the expression of inflammation cytokines at mRNA level. After 36 hours of infection with oncoVV or oncoVV-AVL in (a) PLC/PRF/5 cells, (b) Hep3B cells, (c) SK-HEP-1 cells, and (d) HuH-7 cells. The mRNA levels of IL-6, IL-8 and TNF-α were detected by qRT-PCR assay. Data were showed in mean ± SEM, and experiments were performed in triplicate. (* p < 0.05, ** p < 0.01).

Figure 2

The contents of IL-6, IL-8 and TNF-α in cell supernatant of PLC/PRF/5 and Hep3B cells were measured by ELISA. (a) The content of IL-6. (b) The content of IL-8 (c) The content of TNF-α. Data were showed in mean ± SEM, and experiments were performed in triplicate. (* p < 0.05).

Figure 3

oncoVV-AVL significantly increased the transcriptional activity of AP-1 in HCC but not NF-κB in HCC. Transcriptional activity of AP-1 (a) and NF-κB (b) were detected in PLC/PRF/5, Hep3B and SK-HEP-1 cells after oncoVV or oncoVV-AVL infection, with PBS as a negative control. Data were showed in mean ± SEM, and experiments were performed in triplicate. (* p < 0.05, ** p < 0.01).

Figure 4

oncoVV-AVL significantly enhanced the production of c-Jun, c-Fos and their phosphorylation in PLC/PRF/5 cells. (a) The production of c-Fos and c-Jun proteins in PLC/PRF/5 cells. (b) Quantification of total c-Fos and c-Jun proteins. (c) Quantification of phosphorylated c-Fos and c-Jun proteins. Data were showed in mean ± SEM, and experiments were performed in triplicate. (* p < 0.05).

Figure 5

oncoVV-AVL significantly enhanced the production of c-Jun, c-Fos and their phosphorylation in Hep3B cells. GAPDH was used as the loading control, and PBS was used as the negative control. (a) The production of c-Fos and c-Jun proteins in Hep3B cells. (b) Quantification of total c-Fos and c-Jun proteins. (c) Quantification of phosphorylated c-Fos and c-Jun proteins. Data were showed in mean ± SEM, and experiments were performed in triplicate. (* p < 0.05).

Figure 6

oncoVV-AVL significantly enhanced the production of c-Jun, c-Fos and their phosphorylation in SK-HEP-1 cells. GAPDH was used as the loading control, and PBS was used as the negative control. (a) The production of c-Fos and c-Jun proteins in SK-HEP-1 cells. (b) Quantification of total c-Fos and c-Jun proteins. (c) Quantification of phosphorylated c-Fos and c-Jun proteins. Data were showed in mean ± SEM, and experiments were performed in triplicate. (* p < 0.05).

Figure 7

oncoVV-AVL significantly enhanced the production of c-Jun, c-Fos and their phosphorylation in HuH-7 cells. GAPDH was used as the loading control, and PBS was used as the negative control. (a) The production of c-Fos and c-Jun proteins in HuH-7 cells. (b) Quantification of total c-Fos and c-Jun proteins. (c) Quantification of phosphorylated c-Fos and c-Jun proteins. Data were showed in mean ± SEM, and experiments were performed in triplicate. (* p < 0.05).