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A novel tumor suppressor CECR2 down regulation links glutamine metabolism contributes tumor growth in laryngeal squamous cell carcinoma

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Abstract

Purpose

Glutamine plays an important role in tumor metabolism and progression. This research aimed to find out how Gln exert their effects on laryngeal squamous cell carcinoma (LSCC).

Methods

Cell proliferation was measured by CCK8 and EdU assay, mitochondrial bioenergetic activity was measured by mitochondrial stress tests. Gene expression profiling was revealed by RNA sequencing and validated by RT-qPCR. In LSCC patients, protein expression in tumor and adjacent tissues was examined and scored by IHC staining. RNAi was performed by stably expressed shRNA in TU177 cells. In vivo tumor growth analysis was performed using a nude mouse tumorigenicity model.

Results

Gln deprivation suppressed TU177 cell proliferation, which was restored by αKG supplementation. By transcriptomic analysis, we identified CECR2, which encodes a histone acetyl-lysine reader, as the downstream target gene for Gln and αKG. In LSCC patients, the expression of CECR2 in tumors was lower than adjacent tissues. Furthermore, deficiency of CECR2 promoted tumor cell growth both in vitro and in vivo, suggesting it has tumor suppressor effects. Besides, cell proliferation inhibited by Gln withdrawal could be restored by CECR2 depletion, and the proliferation boosted by αKG supplementation could be magnified either, suggested that CECR2 feedback suppressed Gln and αKG’s effect on tumor growth. Transcriptomic profiling revealed CECR2 regulated the expression of a series of genes involved in tumor progression.

Conclusion

We confirmed the Gln-αKG-CECR2 axis contributes to tumor growth in LSCC. This finding provided a potential therapeutic opportunity for the use of associated metabolites as a potential treatment for LSCC.

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Data availability

The datasets used during the present study are available from the corresponding author upon reasonable request.

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Acknowledgements

The authors would like to express their gratitude to EditSprings (https://www.editsprings.com/) for the expert linguistic services provided.

Funding

This study was supported by National Natural Science Foundation of China (No. 81971240 to Liu F., No. 81770988 and 81970869 to Yi H.L., No. 81770987 to Guan J.); Shanghai Municipal Commission of Science and Technology (No.18DZ2260200). Jinan Science and Technology Development Program (No. 201907018 to Zhang N.) and Shandong Provincial Key Research and Development Program (No. 2017G006037 to Zhang N.)

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Author notes

  1. X. Wang and C. Xu authors contributed equally to this work.

Authors and Affiliations

  1. Department of Otolaryngology-Head and Neck Surgery, Otolaryngology Institute of Shanghai JiaoTong University, Shanghai JiaoTong University Affiliated Sixth People’s Hospital, 600 Yishan Road, Xuhui, 200233, Shanghai, China

    Xiaoting Wang, Chong Xu, Shengming Wang, Weijun Huang, Yuenan Liu, Xiaoxu Zhang, Niannian Li, Zhenfei Gao, Fan Wang, Jian Guan, Hongliang Yi & Feng Liu

  2. Shanghai Key Laboratory of Sleep Disordered Breathing, 600 Yishan Road, Xuhui, 200233, Shanghai, China

    Xiaoting Wang, Chong Xu, Shengming Wang, Weijun Huang, Yuenan Liu, Xiaoxu Zhang, Niannian Li, Zhenfei Gao, Fan Wang, Jian Guan, Hongliang Yi & Feng Liu

  3. Department of Oncology, Jinan Central Hospital, Cheeloo College of Medicine, Shandong University, Jinan, 250013, Shandong, China

    Nan Zhang

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Contributions

X.T.W., H.L.Y., J.G. and F.L. designed experiments. X.T.W., C.X. performed experiments and analyzed data. S.M.W., W.J.H. and Y.N.L. contributed to data collection, data interpretation. N.N.L., Z.F.G., F.W. and N.Z. contributed to protocol development. X.T.W., H.L.Y and F.L. prepared the figures and wrote the manuscript.

Corresponding authors

Correspondence to Hongliang Yi or Feng Liu.

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All authors declare that they have no conflict of interest.

Ethical approval

The experimental protocol was established, according to the ethical guidelines of the Declaration of Helsinki (1964) and was approved by Chinese Clinical Trial Registry. The approval number is ChiCTR2000040554.

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Informed consents were obtained from all patients for the use of their samples.

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12094_2021_2603_MOESM1_ESM.png

Supplementary file1 Supplementary Fig. 1 Expression of relevant genes in single cell RNA(scRNA) sequencing in LSCC. a t‐SNE plot of the LSCC cell clusters in global view and the identification of cell‐class composition. b–f Expression of key enzymes, including IDH1 (b), IDH2 (c), GLS (d), PPAT (e) and GLS2 (f), required for Gln and αKG in LSCC. g–i Expression of candidate genes, CHAC1 (g), ULBP1 (h) and SLFN5 (i), in LSCC. (PNG 514 KB)

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Wang, X., Xu, C., Wang, S. et al. A novel tumor suppressor CECR2 down regulation links glutamine metabolism contributes tumor growth in laryngeal squamous cell carcinoma. Clin Transl Oncol 23, 1942–1954 (2021). https://doi.org/10.1007/s12094-021-02603-y

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