Open Access

ARTICLE

Hepatitis B virus X protein-mediated upregulation of miR-221 activates the CXCL12-CXCR4 axis to promote NKT cells in HBV-related hepatocellular carcinoma

YUE CAO, LIN HU, YISHU TANG^*

Department of Laboratory Medicine, The First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, China

* Corresponding Author: YISHU TANG. Email:

(This article belongs to the Special Issue: Recent Advancement in Cancer Molecular Signaling)

BIOCELL 2023, 47(7), 1537-1548. https://doi.org/10.32604/biocell.2023.027205

Received 20 October 2022; Accepted 01 March 2023; Issue published 21 June 2023

Abstract

Backgrounds: Both hepatitis B virus X protein (HBx) and microRNA-221 (miR-221) have been implicated in the development of hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC). The present study demonstrates that HBx promotes HCC cell proliferation via the C-X-C motif chemokine ligand 12-C-X-C chemokine receptor type 4 (CXCL12-CXCR4) axis. We predict that HBx/miR-221-mediated CXCL12/CXCR4 signaling induces NKT cells to promote HBV-related HCC. Methods: After miR-221 mimic, miR-221 mimic negative control, miR-221 inhibitor, miR-221 inhibitor negative control were transfected into cells, the expression of CXCL12 and miR-221 was detected by qPCR and western blot. Then we constructed a stable HBV-HCC cell line. HBV-HCC cells were injected into the nude mice, thus a HBV-HCC mouse model was constructed. Q-PCR and western blot were used to detect the expression of HBx, miR-221, CXCL12 and CXCR4 in tumor tissues. The expression of CXCL12 was detected by immunohistochemistry, and the expression of CXCR4, CD3 and CD56 was detected by immunofluorescence. The levels of CXCL12, IL-2 and TNF-α in serum of mice were detected by ELISA. Sixty-one patients with HBV-related HCC, 61 patients with HBV-related cirrhosis, 61 patients with chronic hepatitis B (CHB) and 30 healthy people were enrolled. CXCL12, cytokine levels, and clinicopathological parameters were tested. Results: Hepatitis B virus X protein upregulates the expression of miR-221 and CXCL12 in lentivirus (LV5)-HBx-transfected HepG2 cells. HBx protein promotes HepG2 cell proliferation in vitro. HBx protein promoted tumor growth via the miR-221/CXCL12/CXCR4 pathway in a mouse tumor model. HBx protein upregulated natural killer T cell expression via the CXCR4/CXCL12 pathway to promote tumor growth. The data demonstrated a positive correlation between CXCL12 concentration with Cre levels and Child-Pugh scores. CXCL12 had an inferior diagnostic efficiency compared to IL-2 and IL-6 for HBV-related HCC. Conclusions: We present evidence that HBx/miR-221-mediated CXCL12/CXCR4 signaling induces NKT cells to promote HBV-related HCC.

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