Krüppel-like Factor 4 Blocks Hepatocellular Carcinoma Dedifferentiation and Progression through Activation of Hepatocyte Nuclear Factor-6.
OBJECTIVE
Tumor differentiation is a behavioral index for hepatocellular carcinoma (HCC) and a prognostic factor for patients with HCC who undergo orthotopic liver transplantation (OLT). However, the molecular basis for HCC differentiation and prognostic value of the underlying molecules that regulate HCC differentiation are unclear. In this study, we defined a potential driver pathway for HCC differentiation and prognostication.


METHODS
The regulation and function of Krüppel-like factor 4 (KLF4) and hepatocyte nuclear factor-6 (HNF-6) in HCC differentiation was evaluated using human tissues, molecular and cell biology, and animal models, and its prognostic significance was determined according to its impact on patient survival.


RESULTS
There was a direct relationship between the expression levels of KLF4 and HNF6 in HCC. Reduced KLF4 or HNF6 expression correlated with high HCC grade. Poorly differentiated HCC cells had lower expression of KLF4 or HNF6 and differentiation-associated markers than did well-differentiated cells. Elevated KLF4 of HNF6 expression induced differentiation of poorly differentiated hepatoma cells. Mechanistically, KLF4 trans-activated HNF-6 expression. Restored HNF-6 expression upregulated expression of differentiation-associated markers and inhibited HCC cell migration and invasion, whereas HNF-6 knockdown did the opposite. Loss of KLF4 expression in primary HCC correlated with reduced overall survival and shortened relapse-free survival durations after OLT. Combination of KLF4 expression and the Milan criteria improved prognostication for HCC after OLT.


CONCLUSIONS
The dysregulated KLF4/HNF-6 pathway drives dedifferentition and progression of HCC, and KLF4 is a biomarker for accurate prognostication of patients with HCC treated by OLT when integrated with the Milan Criteria.
Department of General Surgery, Shanghai Jiaotong University Shanghai General Hospital, Shanghai, PR China. Department of Gastroenterology, Hepatology and Nutrition, The University of Texas MD Anderson Cancer Center, Houston, Texas.
Hongcheng
H
Sun
Department of Pathology, Shanghai Jiaotong University Shanghai General Hospital, Shanghai, PR China.
Huamei
H
Tang
Department of Oncology and Tumor Institute, Shanghai East Hospital, Tongji University School of Medicine, Shanghai, PR China.
Dacheng
D
Xie
Department of Gastroenterology, Hepatology and Nutrition, The University of Texas MD Anderson Cancer Center, Houston, Texas.
Zhiliang
Z
Jia
Liver Transplantation Section, Center for Organ Transplantation, Fudan University, Shanghai, China.
Zhenyu
Z
Ma
Department of Gastroenterology, Hepatology and Nutrition, The University of Texas MD Anderson Cancer Center, Houston, Texas.
Daoyan
D
Wei
Department of Gastroenterology, Hepatology and Nutrition, The University of Texas MD Anderson Cancer Center, Houston, Texas.
Lopa
L
Mishra
Department of Oncology and Tumor Institute, Shanghai East Hospital, Tongji University School of Medicine, Shanghai, PR China.
Yong
Y
Gao
Department of Pathology Hainan Medical College Affiliated Hospital, Haikou, PR China. kepxie@mdanderson.org zhenghoho@aliyun.com xczmd@sjtu.edu.cn.
Shaojiang
S
Zheng
Department of Gastroenterology, Hepatology and Nutrition, The University of Texas MD Anderson Cancer Center, Houston, Texas. kepxie@mdanderson.org zhenghoho@aliyun.com xczmd@sjtu.edu.cn.
Keping
K
Xie
Department of General Surgery, Shanghai Jiaotong University Shanghai General Hospital, Shanghai, PR China. kepxie@mdanderson.org zhenghoho@aliyun.com xczmd@sjtu.edu.cn.
Zhihai
Z
Peng
D000818:Animals; D006528:Carcinoma, Hepatocellular / Q000235:genetics* / Q000473:pathology*; D002454:Cell Differentiation / Q000235:genetics*; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000235:genetics; D018450:Disease Progression; D018572:Disease-Free Survival; D015972:Gene Expression Regulation, Neoplastic / Q000235:genetics; D056945:Hep G2 Cells; D051559:Hepatocyte Nuclear Factor 6 / Q000235:genetics*; D006801:Humans; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors / Q000235:genetics*; D008113:Liver Neoplasms / Q000235:genetics* / Q000473:pathology*; D051379:Mice; D009364:Neoplasm Recurrence, Local / Q000235:genetics; D011379:Prognosis; D015854:Up-Regulation / Q000235:genetics
D051559:Hepatocyte Nuclear Factor 6; C000715467:KLF4 protein, human; C000715468:Klf4 protein, mouse; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors