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A lectin-based glycomic approach identifies FUT8 as a driver of radioresistance in oesophageal squamous cell carcinoma.
Cellular Oncology ( IF 6.6 ) Pub Date : 2020-05-30 , DOI: 10.1007/s13402-020-00517-5
Li Shen 1, 2 , Min Xia 2 , Xinzhou Deng 1 , Qing Ke 1 , Chuanyi Zhang 1 , Feng Peng 1 , Xiaoxia Dong 3 , Zhiguo Luo 1, 4, 5
Affiliation  

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.


中文翻译:

基于凝集素的糖化方法可将FUT8识别为食道鳞状细胞癌中的放射抗性驱动器。

目的

放射抵抗被认为是食管鳞状细胞癌(ESCC)放射治疗失败的主要因素。据报道,异常细胞表面糖基化与不同类型肿瘤中的抗辐射性相关。但是,尚未确定与ESCC抗辐射性相关的糖基变化和相应的酶。

方法

通过分次照射从亲代ESCC细胞系EC109和TE-1建立了两个抗辐射细胞系EC109R和TE-1R。凝集素微阵列用于筛选改变的聚糖模式。RNA测序(RNA-seq)用于鉴定差异表达的糖基转移酶。使用Cell Counting Kit-8,集落形成和流式细胞术检测细胞活力和放射敏感性。通过免疫组织化学评估ESCC组织中糖基转移酶的表达。使用裸鼠异种移植模型分析体内放射敏感性。使用基于凝集素的下拉分析结合质谱法验证了酶的下游效应子。

结果

我们发现EC109R和TE-1R细胞比亲代EC109和TE-1细胞对辐射的抵抗力更高。使用凝集素微阵列与RNA测序相结合,我们发现α1,6-岩藻糖基转移酶(FUT8)在抗辐射的ESCC细胞系中过表达。功能获得和功能丧失研究均证实FUT8调节ESCC细胞对辐射的敏感性。重要的是,我们发现在接受放射治疗的ESCC患者中,高FUT8表达与放射抵抗和不良预后正相关。此外,FUT8抑制抑制了辐射后裸鼠体内异种移植肿瘤的生长和形成。使用基于凝集素的下拉测定法和质谱,我们发现CD147可以被FUT8糖基化。不出所料

结论

我们的结果表明,FUT8通过针对CD147充当ESCC中的无线电抗性驱动器。因此,FUT8可以作为预测ESCC患者对放射治疗反应的标记。
更新日期:2020-05-30
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