Glucose deprivation–induced aberrant FUT1-mediated fucosylation drives cancer stemness in hepatocellular carcinoma
Jane H.C. Loong, … , Jing-Ping Yun, Stephanie K.Y. Ma
Jane H.C. Loong, … , Jing-Ping Yun, Stephanie K.Y. Ma
Published April 20, 2021
Citation Information: J Clin Invest. 2021;131(11):e143377. https://doi.org/10.1172/JCI143377.
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Research Article Gastroenterology Oncology

Glucose deprivation–induced aberrant FUT1-mediated fucosylation drives cancer stemness in hepatocellular carcinoma

Abstract

Rapidly growing tumors often experience hypoxia and nutrient (e.g., glucose) deficiency because of poor vascularization. Tumor cells respond to the cytotoxic effects of such stresses by inducing molecular adaptations that promote clonal selection of a more malignant tumor-initiating cell phenotype, especially in the innermost tumor regions. Here, we report a regulatory mechanism involving fucosylation by which glucose restriction promotes cancer stemness to drive drug resistance and tumor recurrence. Using hepatocellular carcinoma (HCC) as a model, we showed that restricted glucose availability enhanced the PERK/eIF2α/ATF4 signaling axis to drive fucosyltransferase 1 (FUT1) transcription via direct binding of ATF4 to the FUT1 promoter. FUT1 overexpression is a poor prognostic indicator for HCC. FUT1 inhibition could mitigate tumor initiation, self-renewal, and drug resistance. Mechanistically, we demonstrated that CD147, ICAM-1, EGFR, and EPHA2 are glycoprotein targets of FUT1, in which such fucosylation would consequently converge on deregulated AKT/mTOR/4EBP1 signaling to drive cancer stemness. Treatment with an α-(1,2)-fucosylation inhibitor sensitized HCC tumors to sorafenib, a first-line molecularly targeted drug used for advanced HCC patients, and reduced the tumor-initiating subset. FUT1 overexpression and/or CD147, ICAM-1, EGFR, and EPHA2 fucosylation may be good prognostic markers and therapeutic targets for cancer patients.

Authors

Jane H.C. Loong, Tin-Lok Wong, Man Tong, Rakesh Sharma, Lei Zhou, Kai-Yu Ng, Hua-Jian Yu, Chi-Han Li, Kwan Man, Chung-Mau Lo, Xin-Yuan Guan, Terence K. Lee, Jing-Ping Yun, Stephanie K.Y. Ma

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Figure 5

FUT1 overexpression is important in mediating growth, self-renewal, and drug resistance in HCC cells.

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FUT1 overexpression is important in mediating growth, self-renewal, and...
(A) Validation of FUT1 overexpression or knockdown in Huh7 and CLC13 HCC cells by Western blotting. (B) An in vitro limiting-dilution assay revealed that FUT1 overexpression and knockdown promoted and attenuated the frequency of tumor-initiating cells, respectively (pairwise tests for differences in stem cell frequencies). (C) Annexin V/PI analysis found that FUT1 overexpression augmented apoptosis in Huh7 cells treated with 4 μM sorafenib (72 hours) and CLC13 cells treated with 4 μM sorafenib (48 hours). DMSO (2%) was used as a control (1-way ANOVA). The reverse phenotype was observed in a Huh7 FUT1-knockdown model (4 μM sorafenib, 72 hours). (D) FUT1 overexpression promoted HCC tumor growth in an in vivo subcutaneous implantation model. Representative image of tumors harvested from mice injected with 5000 cells. Scale bar: 1 cm. Kaplan-Meier curve showing a significant decrease in the percentage of tumor-free survival upon FUT1 overexpression (n = 6). In vivo limiting-dilution assays found that FUT1 overexpression enhanced tumor incidence, expedited tumor latency, and increased the frequency of tumor-initiating cells (pairwise tests for differences in stem cell frequencies and Kaplan-Meier survival curve using the log-rank test). The data shown in A–C are representative of 3 independent experiments. NTC, nontargeting control; OE, overexpression. *P < 0.05; **P < 0.01; ***P < 0.001.