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A lectin-based glycomic approach identifies FUT8 as a driver of radioresistance in oesophageal squamous cell carcinoma

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Abstract

Purpose

Radio-resistance is recognized as a main factor in the failure of radiotherapy in oesophageal squamous cell carcinoma (ESCC). Aberrant cell surface glycosylation has been reported to correlate with radio-resistance in different kinds of tumours. However, glycomic alterations and the corresponding enzymes associated with ESCC radio-resistance have not yet been defined.

Methods

Two radioresistant cell lines, EC109R and TE-1R, were established from parental ESCC cell lines EC109 and TE-1 by fractionated irradiation. A lectin microarray was used to screen for altered glycan patterns. RNA-sequencing (RNA-seq) was employed to identify differentially expressed glycosyltransferases. Cell Counting Kit-8, colony formation and flow cytometry assays were used to measure cell viability and radiosensitivity. Expression of glycosyltransferase in ESCC tissues was assessed by immunohistochemistry. In vivo radiosensitivity was analysed using a nude mouse xenograft model. Downstream effectors of the enzyme were verified using a lectin-based pull-down assay combined with mass spectrometry.

Results

We found that EC109R and TE-1R cells were more resistant to irradiation than the parental EC109 and TE-1 cells. Using lectin microarrays combined with RNA sequencing, we found that α1, 6-fucosyltransferase (FUT8) was overexpressed in the radioresistant ESCC cell lines. Both gain- and loss-of-function studies confirmed that FUT8 regulates the sensitivity of ESCC cells to irradiation. Importantly, we found that high FUT8 expression was positively linked to radio-resistance and a poor prognosis in ESCC patients who received radiation therapy. Moreover, FUT8 inhibition suppressed the growth and formation of xenograft tumours in nude mice after irradiation. Using a lectin-based pull-down assay and mass spectrometry, we found that CD147 could be glycosylated by FUT8. As expected, inhibition of CD147 partly reversed FUT8-induced radio-resistance in ESCC cells.

Conclusions

Our results indicate that FUT8 functions as a driver of radio-resistance in ESCC by targeting CD147. Therefore, FUT8 may serve as a marker for predicting the response to radiation therapy in patients with ESCC.

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Acknowledgements

We thank American Journal Experts (AJE) for English language editing.

Funding

This study was funded by the National Natural Science Foundation of China (NO. 81502666, NO. 81902494), the Natural Science Foundation of Hubei Province (NO.2019CFA034, NO. 2019CFB441), the Initial Project for Post-Graduates of the Hubei University of Medicine (NO.2016QDJZR10), the Free Exploration Foundation of the Hubei University of Medicine (NO.FDFR201802), and the Innovation Project for graduates of the Hubei University of Medicine (NO. YC2019007).

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Author notes

  1. Li Shen and Min Xia contributed equally to this work.

Authors and Affiliations

  1. Department of Clinical Oncology, Taihe Hospital, Hubei University of Medicine, Shiyan, Hubei, 442000, People’s Republic of China

    Li Shen, Xinzhou Deng, Qing Ke, Chuanyi Zhang, Feng Peng & Zhiguo Luo

  2. Department of Biochemistry, School of Basic Medical Sciences, Hubei University of Medicine, Shiyan, Hubei, 442000, People’s Republic of China

    Li Shen & Min Xia

  3. Department of Pharmacology, School of Basic Medical Sciences, Hubei University of Medicine, Shiyan, Hubei, 442000, People’s Republic of China

    Xiaoxia Dong

  4. Hubei Provincial Research Center for Precision Medicine of Cancer, Shiyan, Hubei, 442000, People’s Republic of China

    Zhiguo Luo

  5. Hubei Key Laboratory of Embryonic Stem Cell Research, Hubei University of Medicine, Shiyan, Hubei, 442000, People’s Republic of China

    Zhiguo Luo

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  1. Li Shen
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  2. Min Xia
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Contributions

Li Shen and Min Xia designed the study and performed the experiments. Xinzhou Deng, Qing Ke, Chuanyi Zhang and Feng Peng collected and analysed the data. Xiaoxia Dong and Zhiguo Luo prepared the manuscript. All authors read and approved the final manuscript.

Corresponding authors

Correspondence to Xiaoxia Dong or Zhiguo Luo.

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Informed consent was obtained from all patients enrolled in the present study. This study was approved by the Ethics Committee of the Hubei University of Medicine (Hubei, China). All animal experiments were performed according to protocols approved by the Committee on Animal Welfare of the Hubei University of Medicine.

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Shen, L., Xia, M., Deng, X. et al. A lectin-based glycomic approach identifies FUT8 as a driver of radioresistance in oesophageal squamous cell carcinoma. Cell Oncol. 43, 695–707 (2020). https://doi.org/10.1007/s13402-020-00517-5

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