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Hypoxia-induced TUFT1 promotes the growth and metastasis of hepatocellular carcinoma by activating the Ca2+/PI3K/AKT pathway

Abstract

Tuftelin1 (TUFT1), an acidic protein constituent of developing and mineralizing tooth tissues, is regulated by hypoxia and the Hedgehog signaling pathway. We investigated the role of TUFT1 in hepatocellular carcinoma (HCC). qRT-PCR, immunohistochemistry and western blot were employed to evaluate TUFT1 level in HCC. MTT, BrdU, 3D culture and Transwell assays were used to assess cell viability, proliferation, in vitro growth, migration, and invasion. Subcutaneous and tail vein injection models were established to investigate in vivo growth and metastasis. Chromatin immunoprecipitation was performed to assess binding of hypoxia-inducible factor 1α (HIF-1α) to TUFT1 promoter. A microRNA array was used to identify hypoxia-related microRNAs. TUFT1 was elevated in HCC, and correlated with unfavorable clinicopathologic characteristics and poor survival. TUFT1 promoted HCC cell growth, metastasis and epithelial-mesenchymal transition in vitro and in vivo via activation of Ca2+/PI3K/AKT pathway. Hypoxia induced TUFT1 expression in an HIF-1α dependent manner, and TUFT1 expression was positively correlated with HIF-1α level in HCC tissues. Hypoxiaenhanced TUFT1 expression by downregulating miR-671-5p rather than by directly promoting the binding of HIF-1α to TUFT1 promoter. MiR-671-5p interacted with the 3′-UTR of TUFT1 mRNA and subsequently inhibited TUFT1 expression. Consequently, knockdown of TUFT1 blocked the effects of hypoxia in promoting HCC progression. TUFT1 promoted the growth, metastasis and EMT of HCC cells through activating Ca2+/PI3K/AKT pathway. The hypoxic microenvironment increased the expression of TUFT1 via downregulation of miR-671-5p. TUFT1 may function as a potential therapeutic target for the intervention and treatment of HCC.

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Acknowledgements

This study was supported by grants from the National Natural Science Foundation of China (81874049, 81773123, 81572847, 81502092); Innovation Capacity Support Plan in Shaanxi Province of China (2018KJXX-045); the Fundamental Research Funds for the Central Universities (7N010011015).

Funding

This study was supported by grants from the National Natural Science Foundation of China (81874049, 81773123, 81572847, 81502092); Innovation Capacity Support Plan in Shaanxi Province of China (2018KJXX-045); the Fundamental Research Funds for the Central Universities (7N010011015).

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Correspondence to Qingguang Liu or Kangsheng Tu.

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These authors contributed equally: Changwei Dou, Zhenyu Zhou, Qiuran Xu, Zhikui Liu

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Dou, C., Zhou, Z., Xu, Q. et al. Hypoxia-induced TUFT1 promotes the growth and metastasis of hepatocellular carcinoma by activating the Ca2+/PI3K/AKT pathway. Oncogene 38, 1239–1255 (2019). https://doi.org/10.1038/s41388-018-0505-8

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