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MiR-222-3p induced by hepatitis B virus promotes the proliferation and inhibits apoptosis in hepatocellular carcinoma by upregulating THBS1

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Abstract

This study aimed to explore the role of miR-222-3p in hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC). MiR-222-3p expression in tumor tissues of HBV (+) or HBV (−) HCC patients and corresponding cell lines was detected by quantitative reverse transcription PCR (qRT-PCR). Cell proliferation was assessed by cell counting kit-8 (CCK-8) and colony formation assays. Cell apoptosis was evaluated by flow cytometry. The potential targets of miR-222-3p were predicted by Targetscan, and the binding relationship between miR-222-3p and thrombospondin-1 (THBS1) was determined by luciferase reporter assay and RNA immunoprecipitation (RIP) assay. MiR-222-3p was significantly upregulated in HCC tissues and cell lines and further elevated by HBV infection. MiR-222-3p downregulation effectively inhibited the proliferation and induced the apoptosis of HBV (−) HepG2 cells, HBV (+) HepG2.2.15 cells, Huh7-V cells, and Huh7-HBV cells. In addition, miR-222-3p overexpression enhanced the proliferation of these cell lines but exhibited no obvious effect on their apoptosis. Mechanistically, miR-222-3p was directly bound to the 3’-UTR of THBS1 and acted as its competing endogenous RNA (ceRNA). Interestingly, THBS1 silencing attenuated the inhibitory effect of miR-222-3p downregulation on the proliferation of these cell lines in vitro. Our results revealed that HBV infection further increased miR-222-3p expression and promoted HCC progression via miR-222-3p-mediated THBS1 downregulation. Our findings suggest that miR-222-3p might be a potential diagnostic and therapeutic target for HCC and HBV-related HCC.

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The analyzed data sets generated during the study are available from the corresponding author on reasonable request.

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Acknowledgments

The authors would like to express our gratitude to those who have critically reviewed this manuscript and those who give us help during this experiment.

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Authors and Affiliations

  1. Department of Breast and Thyroid Diseases, Shandong Provincial ENT Hospital, Cheeloo College of Medicine, Shandong University, Jinnan, 250022, Shandong, People’s Republic of China

    Yongjie Sun

  2. Department of Breast and Thyroid Surgery, Shandong Provincial Hospital affiliated to Shandong First Medical University, Jinnan, 250022, Shandong, People’s Republic of China

    Peng Shi

  3. Department of Hepatobiliary Surgery, Shandong Provincial Hospital, Cheeloo College of Medicine, Shandong University, Jinnan, 250022, Shandong, People’s Republic of China

    Qi Wu, Bingqiang Liu, Zetao Yu, Hongtao Jia & Hong Chang

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  1. Yongjie Sun
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Contributions

HC: guarantor of integrity of the entire study, study concepts. YS, PS, QW: study design, study perform, original manuscript writing. BL, ZY, HJ: definition of intellectual content, literature research, experimental study, data acquisition.

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Correspondence to Hong Chang.

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This study was approved by the Ethics Committee of the Shandong Provincial Hospital, Cheeloo College of Medicine, Shandong University. All procedures were followed the guideline of this hospital.

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Sun, Y., Shi, P., Wu, Q. et al. MiR-222-3p induced by hepatitis B virus promotes the proliferation and inhibits apoptosis in hepatocellular carcinoma by upregulating THBS1. Human Cell 34, 1788–1799 (2021). https://doi.org/10.1007/s13577-021-00577-1

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