Oncogenic miR-96-5p inhibits apoptosis by targeting the caspase-9 gene in hepatocellular carcinoma

Iwai,Naoto; Yasui,Kohichiroh; Tomie,Akira; Gen,Yasuyuki; Terasaki,Kei; Kitaichi,Tomoko; Soda,Tomohiro; Yamada,Nobuhisa; Dohi,Osamu; Seko,Yuya; Umemura,Atsushi; Nishikawa,Taichiro; Yamaguchi,Kanji; Moriguchi,Michihisa; Konishi,Hideyuki; Naito,Yuji; Itoh,Yoshito

doi:10.3892/ijo.2018.4369

Oncogenic miR-96-5p inhibits apoptosis by targeting the caspase-9 gene in hepatocellular carcinoma

Authors:
- Naoto Iwai
- Kohichiroh Yasui
- Akira Tomie
- Yasuyuki Gen
- Kei Terasaki
- Tomoko Kitaichi
- Tomohiro Soda
- Nobuhisa Yamada
- Osamu Dohi
- Yuya Seko
- Atsushi Umemura
- Taichiro Nishikawa
- Kanji Yamaguchi
- Michihisa Moriguchi
- Hideyuki Konishi
- Yuji Naito
- Yoshito Itoh
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Affiliations: Department of Molecular Gastroenterology and Hepatology, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kyoto 602-8566, Japan
Published online on: April 12, 2018 https://doi.org/10.3892/ijo.2018.4369
Pages: 237-245

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Abstract

The aberrant expression or alteration of microRNAs (miRNAs/miRs) contributes to the development and progression of cancer. In the present study, the functions of miR-96-5p in hepatocellular carcinoma (HCC) were investigated. It was identified that miR-96-5p expression was significantly upregulated in primary HCC tumors compared with their non-tumorous counterparts. A copy number gain was frequently observed at chromosomal region 7q32.2 in which the MIR96 locus is located, suggesting that gene amplification may be one of the mechanisms by which miR-96-5p expression is increased in HCC. Transfection of miR-96-5p mimic into HCC cells decreased the expression of CASP9, which encodes caspase-9, the essential initiator caspase in the mitochondrial apoptotic pathway, at the mRNA and protein levels. A putative binding site for miR-96-5p was identified in the CASP9 3'-untranslated region, and the results of a luciferase assay indicated that CASP9 is a potential direct target of miR-96-5p. The miR-96-5p mimic increased resistance to doxorubicin- and ultraviolet-induced apoptosis through the decrease in caspase-9 expression in HCC cells. Transfection of miR-96-5p inhibitor enhanced the cytotoxic effect of doxorubicin by increasing caspase-9 expression in the HCC cells, suggesting a synergistic effect between the miR-96-5p inhibitor and doxorubicin. In conclusion, the results of the present study suggest that miR-96-5p, which is frequently upregulated in HCC, inhibits apoptosis by targeting CASP9. Therefore, miR-96-5p may be a potential therapeutic target for HCC.

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