青岛大学学报（医学版）2026，Vol.62Issue(3)：385-392,8.DOI:10.11712/jms.2096-5532.2026.62.087
MANF/CK19轴在肝细胞向胆管癌转分化中的调控作用
Regulatory role of the mesencephalic astrocyte-derived neurotrophic factor/cytokeratin 19 axis in hepatocyte-to-cholangiocarcinoma transdifferentiation
摘要
Abstract
Objective To investigate the regulatory role of the mesencephalic astrocyte-derived neurotrophic factor(MANF)/cytokeratin 19(CK19)axis in hepatocyte-to-cholangiocarcinoma-like phenotypic transdifferentiation.Methods The method of thioacetamide(TAA)induction was used to establish a mouse model of intrahepatic cholangiocarcinoma(ICC).Mice were randomly divided into normal control group(Con group),Model group,hepatocyte-specific MANF knockout group(HKO group),and recombinant human MANF intervention group(rhMANF group),with 12 mice in each group.Serum liver function parameters(alanine aminotransferase[ALT],aspartate aminotransferase[AST],total bilirubin[TBIL],and direct bilirubin[DBIL])and MANF were measured,and qRT-PCR,immunohistochemical staining,and immunofluorescent staining were used to measure the expression levels of MANF and CK19 in liver tissue.Primary hepatocytes were isolated and randomly divided into BSA control group and rhMANF treatment group.Cell morphology was observed;Western blot was used to measure the changes in the protein expression levels of MANF,CK19,and hepatocyte nuclear factor 4α(HNF4α);co-immunoprecipitation(Co-IP)was used to observe the interaction between MANF and CK19.Results Compared with the Con group,the Model group had significant increases in the serum levels of ALT,AST,TBIL,DBIL,and MANF and the mRNA and protein expression levels of MANF and CK19 in liver tissue(F=192.48-394.69,P＜0.001).The HKO group had significantly lower levels of the above indicators com-pared with the Model group,while the rhMANF group had significantly higher levels of these indicators compared with the HKO group(P＜0.05).In vitro experiments showed that compared with the BSA group,the rhMANF group showed cholangiocyte-like morphological changes of primary hepatocytes,significant increases in the protein expression levels of MANF and CK19,and a significant reduction in the protein expression level of HNF4α(t=8.97-28.92,P＜0.001).Immunofluorescent staining showed the colocalization of MANF and CK19,and Co-IP further confirmed the interaction between MANF and CK19.Conclusion MANF may drive hepatocytes to acquire a cholangiocyte-like phenotype by interacting with CK19 and synergistically downregulating HNF4α,suggesting that this axis may be one of the important mole-cular pathways involved in the development and progression of ICC.关键词
胶质细胞源性神经营养因子/角蛋白19/胆管肿瘤/肝细胞/细胞转分化Key words
glial cell line-derived neurotrophic factor/keratin-19/bile duct neoplasms/hepatocytes/cell transdifferentiation分类
医药卫生引用本文复制引用
胡炜,刘晗,郝智楠,刘思佚,张庆..MANF/CK19轴在肝细胞向胆管癌转分化中的调控作用[J].青岛大学学报（医学版）,2026,62(3):385-392,8.基金项目
重庆市自然科学基金面上项目(CSTB2024-NSCQ-MSX0251) （CSTB2024-NSCQ-MSX0251）
