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miR-338-3p Inhibits Apoptosis Evasion in Huh7 Liver Cancer Cells by Targeting Sirtuin 6

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Abstract

Liver cancer is one of the most common cancers with an unsatisfactory prognosis and high mortality rate. The ability of liver cancer cells to evade apoptosis results in the poor therapeutic effect of existing treatments and high rates of tumor recurrence and metastasis. This study investigated the effects of abnormal miR-338-3p expression on the ability of liver cancer cells to avoid apoptosis and the mechanism for that avoidance. The levels of miR-338-3p in liver tumor tissues and in different liver cancer cell lines were analysed by qRT-PCR which demonstrated its downregulation in liver tumor tissues and cells, especially in the Huh7 cells. Cell viability and apoptosis rates of Huh7 liver cancer cells were evaluated using the CCK-8 assay, clone formation assay, flow cytometry, and TUNEL assay. whereas the expression levels of apoptosis-related proteins (Bax, caspase-3, Cyt C, and X-linked inhibitor of apoptosis protein [XIAP]) were analyzed by Western blotting. Next, the target relationship between miR-338-3p and Sirtuin 6 (SIRT6) was identified by conducting a dual luciferase reporter gene assay. Overexpression of miR-338-3p was found to inhibit cell viability and promote the apoptosis of Huh7 cells. Additionally, an upregulation of miR-338-3p significantly promoted Bax, caspase-3, and Cyt C expression, and suppressed XIAP and SIRT6 expression. Notably, SIRT6 was proven to be a target gene of miR-338-3p, and SIRT6 overexpression was shown to reverse the anti-tumor effect of miR-338-3p upregulation in Huh7 cells. Furthermore, the apoptosis induction effect of Cisplatin was reduced by SIRT6, but restored by miR-338-3p. In conclusion, this study demonstrated that miR-338-3p increased apoptosis in liver cancer cell Huh7 by inhibiting SIRT6, and thereby enhanced the cytocidal effect of the apoptosis inducer Cisplatin. These results suggest miR-338-3p as a target for treating liver cancer, which might provide a new therapeutic strategy.

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Funding

This research is supported by the Program from Science and Technology (Medical and Health) of Shaoxing (Grant no. 2020A13061 and no. 2018C30025).

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Authors and Affiliations

  1. Department of Oncology, Affiliated Hospital of Shaoxing University, Shaoxing, China

    G. Xiao, Q. Wang, M. Ding, W. Zhu, J. Chang & Y. Fu

  2. Department of General Surgery, Affiliated Hospital of Shaoxing University, Shaoxing, China

    Z. Zhang

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  1. G. Xiao
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  4. Z. Zhang
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  7. Y. Fu
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Contributions

Study design/planning: G.X and Q.W; Data collection/entry: G.X, M.D and Z.Z; Data analysis/statistics: W.Z, J.C and Y.F; Data interpretation: G.X, Q.W and M.D; Preparation of manuscript: G.X and Q.W; Literature analysis/search: G.X and Q.W; Funding collection: G.X. All authors have reviewed the manuscript and consented the submission.

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Correspondence to G. Xiao.

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Ethic statement

This research is agreed by the ethic committee of Affiliated Hospital of Shaoxing University (2020A13061), and informed consent were obtained from the patients.

Conflict of interests

The authors declare that they have no competing interests regarding this research.

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Xiao, G., Wang, Q., Ding, M. et al. miR-338-3p Inhibits Apoptosis Evasion in Huh7 Liver Cancer Cells by Targeting Sirtuin 6. J Evol Biochem Phys 58, 1413–1424 (2022). https://doi.org/10.1134/S002209302205012X

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