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HIF-1α-activated TMEM237 promotes hepatocellular carcinoma progression via the NPHP1/Pyk2/ERK pathway

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Abstract

Background

Hypoxia-inducible factors (HIFs) are the most essential endogenous transcription factors in the hypoxic microenvironment and regulate multiple genes involved in the proliferation, migration, invasion, and EMT of hepatocellular carcinoma (HCC) cells. However, the regulatory mechanism of HIFs in driving HCC progression remains poorly understood.

Methods

Gain- and loss-of-function experiments were carried out to investigate the role of TMEM237 in vitro and in vivo. The molecular mechanisms involved in HIF-1α-induced TMEM237 expression and TMEM237-mediated enhancement of HCC progression were confirmed by luciferase reporter, ChIP, IP-MS and Co-IP assays.

Results

TMEM237 was identified as a novel hypoxia-responsive gene in HCC. HIF-1α directly bound to the promoter of TMEM237 to transactivate its expression. The overexpression of TMEM237 was frequently detected in HCC and associated with poor clinical outcomes in patients. TMEM237 facilitated the proliferation, migration, invasion, and EMT of HCC cells and promoted tumor growth and metastasis in mice. TMEM237 interacted with NPHP1 and strengthened the interaction between NPHP1 and Pyk2 to trigger the phosphorylation of Pyk2 and ERK1/2, thereby contributing to HCC progression. The TMEM237/NPHP1 axis mediates hypoxia-induced activation of the Pyk2/ERK1/2 pathway in HCC cells.

Conclusions

Our study demonstrated that HIF-1α-activated TMEM237 interacted with NPHP1 to activate the Pyk2/ERK pathway, thereby promoting HCC progression.

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Availability of data and materials

All data generated or analyzed during this study are included either in this article or in the supplementary information files.

Abbreviations

HCC:

Hepatocellular carcinoma

HIFs:

Hypoxia-inducible factors

PHD:

Prolyl hydroxylase

HREs:

Hypoxia-responsive elements

TMEM237:

Transmembrane protein 237

JSRDs:

Joubert syndrome-related disorders

EMT:

Epithelial–mesenchymal transition

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Funding

This study was supported by grants from the National Natural Science Foundation of China (82203759), Nature Science Basic Research Program of Shaanxi (2020JC-36), Key Research and Development Program of Shaanxi (2023-YBSF-149), Innovation Capacity Support Plan in Shaanxi Province of China (2023KJXX-107) and Fundamental Research Funds for the Central Universities (xzy012022095).

Author information

Author notes

  1. Tianxiang Chen and Liang Wang contributed equally to this work.

Authors and Affiliations

  1. Department of Hepatobiliary Surgery, The First Affiliated Hospital of Xi’an Jiaotong University, Xi’an, 710061, China

    Tianxiang Chen, Runtian Li, Runkun Liu, Yongshen Niu, Qingguang Liu & Kangsheng Tu

  2. Department of Burn and Plastic Surgery, Shaanxi Provincial People’s Hospital, Xi’an, 710068, China

    Liang Wang

  3. Department of General Surgery, The First Affiliated Hospital of Xi’an Medical University, Xi’an, 710077, China

    Chao Chen

  4. The 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, 310014, China

    Ning Zhu

  5. Department of Biochemistry and Molecular Biology, School of Basic Medical Science, Xi’an Jiaotong University Health Science Center, Xi’an, 710061, China

    Zhengtao Xiao

  6. Department of Medical Equipment, Shaanxi Provincial People’s Hospital, Xi’an, 710068, China

    Hui Liu

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  1. Tianxiang Chen
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Contributions

KT and QL conceived and designed the experiments; TC, LW, CC, RL, NZ, RL, and YN performed the experiments; TC, HL and ZX analyzed the data; HL and ZX contributed reagents/materials/analysis tools; TC and KT wrote the paper. All authors read and approved the final manuscript.

Corresponding authors

Correspondence to Qingguang Liu or Kangsheng Tu.

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Conflict of interest

The authors declare that they have no competing interests.

Ethics approval and consent to participate

All procedures performed in studies involving human participants were in accordance with the ethical standards of the Research Ethics Committee of The First Affiliated Hospital of Xi’an Jiaotong University and with the 1964 Helsinki declaration and its later amendments. All written informed consent to participate in the study was obtained from HCC patients for samples to be collected from them.

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Supplementary Information

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Supplementary file1 (DOCX 16 KB)

Supplementary file2 (DOCX 17 KB)

Supplementary file3 (DOCX 17 KB)

18_2023_4767_MOESM4_ESM.tif

Supplementary Figure 1 The correlations of TMEM237 expression with HIF-1α and TMEM237 CNV in HCC. (A) TCGA data analysis indicated that TMEM237 CNV was positively correlated with TMEM237 expression in HCC tissues. (B) TCGA data analysis demonstrated that HIF-1α was positively correlated with TMEM237 expression in HCC tissues. (C) TCGA data analysis revealed that HIF-1α was not associated with TMEM237 CNV in HCC tissues. (D) The proportion of TCGA-LIHC samples with TEME237 copy gain or loss. (E) A positive correlation between HIF-1α and TMEM237 expression was observed in HCC samples with TMEM237 copy gains. (F) A positive correlation between HIF-1α and TMEM237 expression was detected in HCC samples without TMEM237 copy gains (TIF 373 KB)

18_2023_4767_MOESM5_ESM.tif

Supplementary Figure 2 The expression and clinical significance of TMEM237 in HCC. (A) The difference in TMEM237 expression between HCC and nontumor tissues from the GEO database (GSE45436). (B and C) TCGA data were analyzed to determine TMEM237 expression in HCC tissues with different tumor grades and stages. (D) TCGA data analysis confirmed that TMEM237 CNV was not associated with the prognosis of HCC patients (TIF 153 KB)

18_2023_4767_MOESM6_ESM.tif

Supplementary Figure 3 The differentially expressed genes in HCCLM3 cells with or without TMEM237 knockdown. HCCLM3 cells that were transfected with shNC or shTMEM237 were analyzed by RNA-seq. (A) Volcano map of differentially expressed genes in HCCLM3 cells with or without TMEM237 knockdown. (B) Heatmap of differentially expressed genes in HCCLM3 cells with or without TMEM237 knockdown (TIF 263 KB)

Supplementary Figure 4 Modulation of TMEM237 expression in HCC cells (TIF 153 KB)

18_2023_4767_MOESM8_ESM.tif

Supplementary Figure 5 TMEM237 promotes the proliferation and invasion of HCC cells. (A) Colony formation, (B) EdU, and (C) Transwell assays were conducted to evaluate the effects of TMEM237 overexpression and knockdown on the proliferation, migration and invasion of HCC cells. Scale bar: 50 μm for the EdU results and 200 μm for the transwell results (TIF 6157 KB)

Supplementary Figure 6 Mass spectrometry analysis of NPHP1 protein (TIF 223 KB)

Supplementary Figure 7 Proteins interacting with MAPK1 (TIF 4686 KB)

18_2023_4767_MOESM11_ESM.tif

Supplementary Figure 8 The NPHP1/Pyk2/ERK1/2 pathway mediates the biological function of TMEM237 in HCC cells. (A) Colony formation, (B) EdU, and (C) transwell assays were employed to examine the proliferation, migration, and invasion of HCC cells in the indicated groups. Scale bar: 50 μm for the EdU results and 200 μm for the transwell results (TIF 8630 KB)

Supplementary Figure 9 The correlations between TMEM237 and HIF-1α, p-Pyk2 and p-ERK1/2 in HCC tissues (TIF 91 KB)

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Chen, T., Wang, L., Chen, C. et al. HIF-1α-activated TMEM237 promotes hepatocellular carcinoma progression via the NPHP1/Pyk2/ERK pathway. Cell. Mol. Life Sci. 80, 120 (2023). https://doi.org/10.1007/s00018-023-04767-y

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