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Claudin-1/4 as directly target gene of HIF-1α can feedback regulating HIF-1α by PI3K-AKT-mTOR and impact the proliferation of esophageal squamous cell though Rho GTPase and p-JNK pathway

Cancer Gene Therapy volume 29, pages 665–682 (2022)Cite this article

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Abstract

Immunohistochemical microarray comprising 80 patients with esophageal squamous cell carcinoma (ESCC) and discovered that the expression of CLDN1 and CLDN4 were significantly higher in cancer tissues compared to para-cancerous tissues. Furthermore, CLDN4 significantly affected the overall survival of cancer patients. When two ESCC cell lines (TE1, KYSE410) were exposed to hypoxia (0.1% O2), CLDN1/4 was shown to influence the occurrence and development of esophageal cancer. Compared with the control culture group, the cancer cells cultured under hypoxic conditions exhibited obvious changes in CLDN1 and CLDN4 expression at both the mRNA and protein levels. Through genetic intervention and Chip, we found that HIF-1α could directly regulate the expression of CLDN1 and CLDN4 in cancer cells. Hypoxia can affect the proliferation and apoptosis of cancer cells by regulating the PI3K-Akt-mTOR pathway. Molecular analysis further revealed that CLDN1 and CLDN4 can participate in the regulation process and had a feedback regulatory effect on HIF-1α expression in cancer cells. In vitro cellular experiments and vivo experiments in nude mice further revealed that changes in CLDN4 expression in cancer cells could affect the proliferation of cancer cells via regulation of Rho GTP and p-JNK pathway. Whether CLDN4 can be target for the treatment of ESCC needs further research.

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Fig. 1: CLDN1/4 expression analysis in ESCC patients.
Fig. 2: 2 cancer cell lines were treated with 0.1% O2.
Fig. 3: HIF-1α can directly regulate the CLDN1/4 protein expression in ESCC cell lines.
Fig. 4: Effect of hypoxic culture on cell apoptosis of cancer cells.
Fig. 5: Interfering with the expression of CLDN1/4 in cancer cells can affect the expression of p-AKT and mTOR proteins and feedback regulate HIF-1α in normoxia condition.
Fig. 6: Effect of hypoxic culture on cell proliferation and cell viability of cancer cells.
Fig. 7: CLDN1 and CLDN4 affect the proliferation of cancer cells though Rho GTPase and p-JNK pathway.
Fig. 8: CLDN1 and CLDN4 can affect the proliferation of cancer cells in vivo experiment.

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Acknowledgements

We are grateful to Darren Edwards for comments on the paper. The present study was supported by grants from the National Natural Science Foundation of China (grant No. 81902763) and Major Science and Technology Plan Project of Hainan Province (grant No. ZDKJ202005).

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Authors and Affiliations

  1. Department of Otolaryngology-Head and Neck Surgery, Shandong Provincial ENT Hospital, Cheeloo College of Medicine, Shandong University, Jinan, 250022, P.R. China

    Hong Liu, Shenli Zhou, Xianfang Liu & Wei Xu

  2. Department of Otolaryngology, The Fourth Hospital of Jinan, Jinan, Shandong, 250031, China

    Zhancheng Zhang

  3. Department of Otolaryngology, Shanxi Provincial People’s Hospital Affiliated to Shanxi Medical University, Taiyuan, 038000, P.R. China

    Guodong Li

  4. School of Dentistry, Cardiff University, Cardiff, CF14 4XY, UK

    Bing Song

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Liu, H., Zhang, Z., Zhou, S. et al. Claudin-1/4 as directly target gene of HIF-1α can feedback regulating HIF-1α by PI3K-AKT-mTOR and impact the proliferation of esophageal squamous cell though Rho GTPase and p-JNK pathway. Cancer Gene Ther 29, 665–682 (2022). https://doi.org/10.1038/s41417-021-00328-2

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