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MiR-375 Impairs the Invasive Capabilities of Hepatoma Cells by Targeting HIF1α Under Hypoxia

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Abstract

Background and Aims

Hypoxia represents one of the most pervasive microenvironmental stresses in HCC due to the overwhelming growth and inadequate blood supply. HIF1α as an important transcription factor participates in the regulation of various biological behaviors of HCC cells under hypoxia. Our previous study indicated that miR-375 is a hypoxia-associated miRNA. However, the interaction between miR-375 and HIF1α remains unclear.

Methods

Bioinformatic analysis was performed for miRNA screening. qRT-PCR, western blotting, and immunohistochemical staining were used to detect the expression of related molecules. Bioinformatic analysis and dual luciferase assay were used to predict and further confirm the target association. Transwell chamber assay and flow cytometry were, respectively, used to detect migration, invasion and apoptosis of hepatoma cells.

Results

MiR-375 presented an obviously differential expression in human HCCs versus background livers (BLs) and HCCs versus normal liver tissues (NLTs). In rat models, miR-375 was gradually declined during hepatocarcinogenesis. HIF1α was remarkably upregulated at protein level rather than at mRNA level in human HCCs versus BLs, HCCs versus NLTs, BLs versus NLTs, and in rat fibrotic livers versus NLTs. HIF1α was determined to be a target of miR-375. MiR-375 inhibitor induced the migration and invasive capabilities and attenuated apoptosis of hepatoma cells under hypoxia. Depriving HIF1α by siRNA could partially reverse the function of miR-375 inhibitor under hypoxia.

Conclusions

MiR-375 impairs the invasive capabilities of HCC cells by targeting HIF1α under hypoxia.

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Abbreviations

HCC:

Hepatocellular carcinoma

HIF1α:

Hypoxia-inducible factor 1α

miRNAs:

MicroRNAs

PHHC:

Primary human hepatocyte

BL:

Background liver

NLT:

Normal liver tissue

3′UTR:

3′ untranslated region

ATG7:

Autophagy-related protein 7

HIGD1A:

Hypoxia-induced gene domain protein-1a

HRE:

Hypoxia response element

Bax:

Bcl-2-associated X protein

Bcl-2:

B-cell lymphoma-2

CCl4:

Carbon tetrachloride

NAFLD:

Nonalcoholic fatty liver disease

TCGA:

The cancer genome atlas

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Acknowledgment

This work was supported by research grants from the National Natural Science Foundation of China (Nos. 81670554, 81302112, 81870390); the Natural Science Fund for Distinguished Young Scholar of Hubei Province (No. 2017CFA068); the National Key R&D Program of China (No. 2017YFC0112302); and the Ministry of Public Health in Hubei Province of China (No. XF2012-8).

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Authors and Affiliations

  1. Department of Gastroenterology, Zhongnan Hospital of Wuhan University, Wuhan, 430071, China

    Chun Wang, Jie Luo, Mingliang Ye, Yinghui Hong, Jialiang Liu, Jiayan Nie, Qiu Zhao & Ying Chang

  2. Hubei Clinical Center and Key Laboratory of Intestinal and Colorectal Diseases, Wuhan, China

    Chun Wang, Jie Luo, Mingliang Ye, Yinghui Hong, Jialiang Liu, Jiayan Nie, Qiu Zhao & Ying Chang

  3. Department of Coloproctological Surgery, The First Affiliated Hospital, Guangxi Medical University, Nanning, China

    Zhihang Chen

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  1. Chun Wang
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  2. Jie Luo
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  4. Mingliang Ye
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  5. Yinghui Hong
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  9. Ying Chang
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Correspondence to Ying Chang.

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All authors have no financial or non-financial conflicts of interest.

Ethical approval

The human samples and animal experiments used in this study were approved by the Local Research Ethics Committee at the Zhongnan Hospital of Wuhan University (No. 2018078) and the Animal Care and Use Committee of Wuhan University (No. 2017055), respectively.

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Wang, C., Luo, J., Chen, Z. et al. MiR-375 Impairs the Invasive Capabilities of Hepatoma Cells by Targeting HIF1α Under Hypoxia. Dig Dis Sci 66, 493–502 (2021). https://doi.org/10.1007/s10620-020-06202-9

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