CTCF Affects Self-renewal and Immune Escape of Hepatocellular  Carcinoma Stem Cells Through SALL3/DNMT3A Axis

doi:10.3870/j.issn.1672-0741.25.11.015

Abstract

Abstract: Objective To investigate the effects of CTCF on self-renewal and immune escape of liver cancer stem cells through the SALL3/DNMT3A axis.Methods HCCLM3 cells and liver cancer stem cells(LCSCs)were assigned to the HCCLM3 group and LCSC group,respectively.LCSCs transfected with sh-CTCF delivered by tLyP-1-modified extracellular vesicles were defined as the tLyP-1-EV-sh-CTCF-LCSC group.LCSCs treated with the WZB117 inhibitor were defined as the WZB117 inhibitor-LCSC group.Quantitative reverse transcription PCR(qRT-PCR)was used to detect CTCF mRNA expression in the HCCLM3 and LCSC groups,as well as the mRNA expression levels of glycolysis-related markers(HK2 and PFKM)and stemness-related markers(FBP1 and PCK1)in the LCSC,tLyP-1-EV-sh-CTCF-LCSC,and WZB117 inhibitor-LCSC groups.Cell migration and colony formation assays were performed to evaluate the effects of CTCF knockdown and glycolysis inhibition on LCSC stemness.qRT-PCR was further used to assess the effects of CTCF knockdown on SALL3 and DNMT3A expression and the effect of SALL3 overexpression on DNMT3A expression.Results Compared with the untreated control group,cells transfected with tLyP-1-EV-sh-CTCF showed significantly reduced CTCF mRNA and protein expression levels(both P<0.05).Compared with the LCSC group,the tLyP-1-EV-sh-CTCF-LCSC group showed decreased mRNA expression of PFKM and PCK1(both P<0.05),while HK2 and FBP1 showed downward trends without statistical significance(both P>0.05).After WZB117 treatment,the mRNA expression levels of PFKM and PCK1 were also reduced(both P<0.05),whereas HK2 and FBP1 showed decreasing trends without statistical significance(both P>0.05).SALL3 overexpression inhibited PD-L1 expression,promoted MHC-I expression,suppressed LCSC self-renewal,and reduced immune escape-related molecular phenotypes,accompanied by downregulation of glycolysis-related molecules(all P<0.05).Transfection with tLyP-1-EV-sh-CTCF upregulated SALL3 expression and downregulated DNMT3A expression,accompanied by decreased expression of glycolysis-related molecules(all P<0.05).Conclusion CTCF knockdown is associated with downregulation of glycolysis-related markers through modulation of the SALL3/DNMT3A axis,accompanied by attenuated self-renewal and immune escape-related molecular phenotypes in liver cancer stem cells.

Key words: CTCF, SALL3/DNMT3A axis, hepatocellular carcinoma stem cell, self-renewal, immune evasion

CLC Number:

R735.7

Zhu Heng, Lyu Huijuan, Wang Yongrui, et al. CTCF Affects Self-renewal and Immune Escape of Hepatocellular Carcinoma Stem Cells Through SALL3/DNMT3A Axis[J]. Acta Medicinae Universitatis Scientiae et Technologiae Huazhong, 2026, 55(4): 496-503.

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CTCF Affects Self-renewal and Immune Escape of Hepatocellular Carcinoma Stem Cells Through SALL3/DNMT3A Axis

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