Publication: Hsa_circ_0048674 facilitates hepatocellular carcinoma progression and natural killer cell exhaustion depending on the regulation of miR-223-3p/PDL1
Authors
Li, Suihui ; Chen, Zhuangzhong ; Zhou, Ruisheng ; Wang, Sisi ; Wang, Wenping ; Liu, De ; Li, Mengquan ; Guo, Tiansheng
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Publisher
Universidad de Murcia, Departamento de Biologia Celular e Histiologia
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DOI
https://doi.org/10.14670/HH-18-440
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info:eu-repo/semantics/article
Description
Abstract
Background. Circular RNAs (circRNAs)
play vital regulatory roles in human cancers, including
hepatocellular carcinoma (HCC). In this study, we aimed
to explore the functions of hsa_circ_0048674 in HCC
development.
Methods. Quantitative real-time polymerase chain
reaction (qRT-PCR) assay was used to detect
hsa_circ_0048674, ubiquitin-like with PHD and RING
finger domains 1 (UHRF1), microRNA-223-3p (miR223-3p) and programmed death ligand 1 (PDL1). RNase
R assay and Actinomycin D assay were employed to
analyze the stability of hsa_circ_0048674. Cell Counting
Kit-8 (CCK-8) assay, colony formation assay and 5-
ethynyl-2'- deoxyuridine (EdU) assay were conducted to
assess cell proliferation. Flow cytometry analysis,
transwell assay and tube formation assay were carried
out for cell apoptosis, migration, invasion and
angiogenesis, respectively. Western blot assay was
adopted for protein levels. Dual-luciferase reporter assay
and RNA immunoprecipitation (RIP) assay were used to
analyze the relationship between miR-223-3p and
hsa_circ_0048674 or PDL1. Murine xenograft model
assay was conducted for the function of
hsa_circ_0048674 in vivo. Immunohistochemistry (IHC)
assay was used to detect Ki-67 level in tumor tissues.
Enzyme linked immunosorbent assay (ELISA) kits were
employed for the concentrations of inflammatory
factors.
Results. Hsa_circ_0048674 was highly expressed in
HCC tissues and cells. Silencing of hsa_circ_0048674
repressed cell growth, migration, invasion and
angiogenesis and promoted apoptosis in HCC cells in
vitro and hampered tumor growth in vivo.
Hsa_circ_0048674 served as an miR-223-3p sponge to
alter PDL1 expression. MiR-223-3p inhibition or PDL1
overexpression restored the impacts of hsa_circ_
0048674 silencing on HCC malignant behaviors. In
addition, hsa_circ_0048674 knockdown promoted
natural killer (NK) cell-mediated cytotoxicity to HCC
cells.
Conclusion. Hsa_circ_0048674 knockdown
decelerated HCC progression through the mediation of
the miR-223-3p/PDL1 axis.
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Citation
Histology and Histopathology Vol. 37, nº12 (2022)
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