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Rab11-FIP4 interacts with ARF5 to promote cancer stemness in hepatocellular carcinoma

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Abstract

Recent studies suggest that Rab11-family interacting proteins (Rab11-FIPs) play an important role in tumorigenesis and progression. Among the Rab11-FIPs, Rab11-FIP4 has been reported to be significantly upregulated in various cancers, including hepatocellular carcinoma (HCC). However, the possible effect on HCC stemness and the underlying mechanism has never been characterized. Here, we found that Rab11-FIP4 was dramatically increased in HCC cell lines and tissues, and had a positive correlation with cancer stemness. Functional studies revealed that elevated expression of Rab11-FIP4 in HCC cells significantly promoted sphere formation, and enhanced the mRNA and protein levels of stemness-associated markers, ALDH1A1, CD133, NANOG, and OCT4. Conversely, the knockdown of Rab11-FIP4 suppressed the cancer stem cell (CSC)-like characteristics of HCC cells. Moreover, silencing of Rab11-FIP4 obviously increased the sensitivity of HCC cells to sorafenib. Mechanistically, Rab11-FIP4 was shown to interact with ADP-ribosylation factor 5 (ARF5) to influence cell cycle-related proteins, CDK1/cyclin B, thereby promoting HCC stemness. Taken together, our results uncovered an essential role for Rab11-FIP4 in regulating CSC-like features of HCC cells and identified Rab11-FIP4 as a potential target for HCC therapy.

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Data availability

The data used to support the findings of this study are available from the corresponding author upon request.

Abbreviations

NANOG:

Nanog homeobox

SOX2:

SRY-Box transcription factor 2

KLF4:

KLF transcription factor 4

OCT4:

POU domain, class 5, transcription factor 1

ERK1:

Mitogen-activated protein kinase 3

ERK2:

Mitogen-activated protein kinase 1

AKT:

AKT serine/threonine kinase

PRAS40:

AKT1 substrate 1

CDK1:

Cyclin-dependent kinase 1

DMEM:

Dulbecco’s modified Eagle medium

MEM:

Minimum essential medium

RPMI-1640:

Roswell Park Memorial Institute 1640

FBS:

Fetal bovine serum

RT-qPCR:

Quantitative reverse transcription-polymerase chain reaction

GAPDH:

Glyceraldehyde-3-phosphate dehydrogenase

RIPA:

Radio immunoprecipitation assay

CCK-8:

Cell counting kit-8

DMEM/F12:

Dulbecco’s modified Eagle media: nutrient mixture F-12

IgG:

Immunoglobulin G

RT:

Room temperature

SD:

Standard deviation

CD133:

Prominin 1

ALDH1A1:

Aldehyde dehydrogenase 1 family member A1

DYTN:

Dystrotelin

EXOC7:

Exocyst Complex Component 7

MYO1D:

Myosin ID

GO:

Gene ontology

KEGG:

Kyoto Encyclopedia of Genes and Genomes

CDK1:

Cyclin-dependent kinase 1

CCNB1:

Cyclin B1

CCNB1:

Cyclin B2

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Funding

This study was supported by grants from the National Natural Science Foundation of China (No. 82003853), Zhejiang Province Natural Science Foundation of China (No. LQ20H310005 and No. LYY21H310009), Medical and Health Research Program of Zhejiang (No. 2021KY046 and No. 2022KY047), and Leading Talent of “Ten Thousand Plan”—National High-Level Talents Special Support Plan and the Science Technology Plan Project of Zhejiang Province (No. 2020R52029).

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Authors and Affiliations

  1. Center for Clinical Pharmacy, Cancer Center, Department of Pharmacy, Zhejiang Provincial People’s Hospital, Affiliated People’s Hospital, Hangzhou Medical College, Hangzhou, China

    Feifeng Song, Qi Zhang, Xixuan Lu, Tong Xu, Qing Hu, Xiaoping Hu, Yiwen Zhang & Ping Huang

  2. Key Laboratory of Endocrine Gland Diseases of Zhejiang Province, Hangzhou, China

    Feifeng Song, Yiwen Zhang & Ping Huang

  3. Cancer Center, Key Laboratory of Tumor Molecular Diagnosis and Individualized Medicine of Zhejiang Province, Zhejiang Provincial People’s Hospital, Affiliated People’s Hospital, Hangzhou Medical College, Hangzhou, China

    Weijiao Fan

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  1. Feifeng Song
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Contributions

P. H. and F. S. designed the study. P. H. and Y. Z. supervised the study. F. S., Q. Z., X. L., and T. X. jointly performed the experiments. Q. H., X. H., and W. F. analyzed the data. F. S. wrote the manuscript. P. H. and Y. Z. revised the manuscript. All authors read and approved the final manuscript. The authors declare that all data were generated in-house and that no paper mill was used.

Corresponding authors

Correspondence to Yiwen Zhang or Ping Huang.

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The study was approved by the Ethics Committee of Zhejiang Provincial People’s Hospital (approved code NO. KT2022041).

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The authors declare no competing interests.

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Key points

• RAB11-FIP4 promotes stemness and increases sorafenib resistance in HCC.

• Rab11-FIP4 interacts with ARF5 to promote stemness in HCC.

• Rab11-FIP4 promotes HCC stemness via influencing the CDK1/cyclin B complex.

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Song, F., Zhang, Q., Lu, X. et al. Rab11-FIP4 interacts with ARF5 to promote cancer stemness in hepatocellular carcinoma. J Physiol Biochem 79, 757–770 (2023). https://doi.org/10.1007/s13105-023-00972-2

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