SPTBN1 inhibits inflammatory responses and hepatocarcinogenesis via the stabilization of SOCS1 and downregulation of p65 in hepatocellular carcinoma

Abstract

Background: Spectrin, beta, non-erythrocytic 1 (SPTBN1), an adapter protein for transforming growth factor beta (TGF-β) signaling, is recognized as a tumor suppressor in the development of hepatocellular carcinoma (HCC); however, the underlying molecular mechanisms of this tumor suppression remain obscure. Methods: The effects on expression of pro-inflammatory cytokines upon the inhibition or impairment of SPTBN1 in HCC cell lines and liver tissues of Sptbn1 +/- and wild-type (WT) mice were assessed by analyses of quantitative real-time reverse-transcription polymerase chain reaction (QRT-PCR), enzyme linked immunosorbent assay (ELISA), Western blotting and gene array databases from HCC patients. We investigated the detailed molecular mechanisms underlying the inflammatory responses by immunoprecipitation-Western blotting, luciferase reporter assay, chromatin immunoprecipitation quantitative real time PCR (ChIP-qPCR), immunohistochemistry (IHC) and electrophoretic mobility shift assay (EMSA). The proportion of myeloid-derived suppressor cells in liver, spleen, bone marrow and peripheral blood samples from WT and Sptbn1 +/- mice were measured by fluorescence-activated cell sorting (FACS) analysis. Further, the hepatocacinogenesis and its correlation with inflammatory microenvironment by loss of SPTBN1/SOCS1 and induction of p65 were analyzed by treating WT and Sptbn1 +/- mice with 3,5-diethoxycarbonyl-1,4-dihydrocollidine (DDC). Results: Loss of SPTBN1 in HCC cells upregulated the expression of pro-inflammatory cytokines including interleukin-1α (IL-1α), IL-1β, and IL-6, and enhanced NF-κB transcriptional activation. Mechanistic analyses revealed that knockdown of SPTBN1 by siRNA downregulated the expression of suppressor of cytokine signaling 1 (SOCS1), an E3 ligase of p65, and subsequently upregulated p65 accumulation in the nucleus of HCC cells. Restoration of SOCS1 abrogated this SPTBN1 loss-associated elevation of p65 in HCC cells. In human HCC tissues, SPTBN1 gene expression was inversely correlated with gene expression of IL-1α, IL-1β and IL-6. Furthermore, a decrease in the levels of SPTBN1 gene, as well as an increase in the gene expression of IL-1β or IL-6 predicted shorter relapse free survival in HCC patients, and that HCC patients with low expression of SPTBN1 or SOCS1 protein is associated with poor survival. Heterozygous loss of SPTBN1 ( Sptbn1 +/- ) in mice markedly upregulated hepatic expression of IL-1α, IL-1β and IL-6, and elevated the proportion of myeloid-derived suppressor cells (MDSCs) and CD4 + CD25 + Foxp3 + regulatory T cells (Foxp3 + Treg) cells in the liver, promoting hepatocarcinogenesis of mouse fed by DDC. Conclusions : Our findings provided evidence ...

Ferroptosis Driver SOCS1 and Suppressor FTH1 Independently Correlate With M1 and M2 Macrophage Infiltration in Head and Neck Squamous Cell Carcinoma

“…For example, a study of 83 patients with esophageal cancer found that exosome-associated miR-19b-3p promotes tumor progression by inhibiting SOCS1 expression (Deng et al, 2021). Moreover, loss of SPTBN1 expression induces liver cancer through downregulation of SOCS1 expression as well (Lin et al, 2021). In nasopharyngeal carcinoma, LINC00669 protects SOCS1 from ubiquitinating STAT1, which promotes cancer cell proliferation and invasion (Qing et al, 2020).…”

Identifying In Vitro Cultured Human Hepatocytes Markers with Machine Learning Methods Based on Single-Cell RNA-Seq Data

“…The high expression of NRAGE facilitates the repair of homologous recombination and can make cells radioresistant by altering subcellular localization ( Xue et al, 2010 ; Chang et al, 2018 ; Liu et al, 2020 ). The high expression of SPTBN1 can suppress inflammation in the liver ( Lin et al, 2021 ). Our rule demonstrated the specificity of the function of hepatocytes differentiated from different original cells, proving the superiority of our method.…”

Identification of Therapeutic Targets for the Selective Killing of HBV-Positive Hepatocytes

“…ARRB1 has been indicated to promote HCC, and was upregulated by hepatitis B virus X protein (HBx) in mouse models [ 59 ]. Interestingly, among other candidate genes, some genes are related to the progression of HCC, including, GATAD1 [ 44 , 45 ], FOXK1 [ 46 ], RALGAPA2 [ 47 ], PDE4A [ 48 ], TRIP6 [ 49 ], SPTBN1 [ 53 ], PTP4A3 [ 54 ], TRIM4 [ 56 ], and LMNB1 [ 60 ]. These results suggest that the 36 candidate genes are associated with the functions of HBV and hepatocytes; however, this association needs to be further validated.…”