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N-Hydroxyphthalimide exhibits antitumor activity by suppressing mTOR signaling pathway in BT-20 and LoVo cells
Min Wang1,2, Ankun Zhou3, Tao An4,5
1State Key Laboratory of Phytochemistry and Plant Resources in West China, Kunming Institute of Botany, Chinese Academy of Sciences, Kunming, 650201, China. wangmin1@mail.kib.ac.cn.
Background:
N-Hydroxyphthalimide (NHPI), an important chemical raw material, was found to have potent and selective anti-proliferative effect on human breast carcinoma BT-20 cells, human colon adenocarcinoma LoVo and HT-29 cells during our screening for anticancer compounds. The purpose of this study is to assess the antitumor efficacy of NHPI in vitro and in vivo and to explore the underlying antitumor mechanism.
Methods:
Cell cytotoxicity of NHPI was evaluated using MTS assay and cell morphological analysis. After NHPI treatment, cell cycle, apoptosis and mitochondrial membrane potential were analyzed using flow cytometer. The subcellular localization of eukaryotic initiation factor 4E (eIF4E) was analyzed by immunofluorescence assay. The antitumor efficacy of NHPI in vivo was tested in BT-20 xenografts. The underlying antitumor mechanisms of NHPI in vitro and in vivo were investigated with western blot analysis in NHPI-treated cancer cells and tumor tissues. Statistical significance was determined using Student's t-test.
Results:
In vitro, NHPI selectively inhibited the proliferation and induced G2/M phase arrest in BT-20 and LoVo cells, which was attributed to the inhibition of cyclin B1 and cdc2 expressions. Furthermore, NHPI induced apoptosis via mitochondrial pathway. Of note, NHPI effectively inhibited mammalian target of rapamycin (mTOR) complex 1 (mTORC1) and mTOR complex 2 (mTORC2) signaling, and overcame the feedback activation of Akt and extracellular signal-regulated kinase (ERK) caused by mTORC1 inhibition in BT-20 and LoVo cells. In vivo, NHPI inhibited tumor growth and suppressed mTORC1 and mTORC2 signaling in BT-20 xenografts with no obvious toxicity.
Conclusions:
We found for the first time that NHPI displayed antitumor activity which is associated with the inhibition of mTOR signaling pathway. Our findings suggest that NHPI may be developed as a promising candidate for cancer therapeutics by targeting mTOR signaling pathway and as such warrants further exploration.
Insights
N-Hydroxyphthalimide (NHPI) shows antitumor effects by inhibiting cancer cell proliferation and inducing apoptosis. This compound targets the mTOR signaling pathway, suggesting its potential as a novel cancer therapeutic.
Area of Science:
- Biochemistry
- Molecular Biology
- Pharmacology
Background:
- N-Hydroxyphthalimide (NHPI) exhibits selective anti-proliferative effects on human breast carcinoma BT-20 and colon adenocarcinoma LoVo and HT-29 cells.
- NHPI is a chemical raw material with potential anticancer properties identified during compound screening.
Purpose of the Study:
- To evaluate the in vitro and in vivo antitumor efficacy of NHPI.
- To elucidate the underlying antitumor mechanisms of NHPI.
Main Methods:
- Cell cytotoxicity, cell cycle, apoptosis, and mitochondrial membrane potential were assessed using MTS assay and flow cytometry.
- Western blot analysis and immunofluorescence were used to investigate molecular mechanisms and protein expression.
- In vivo antitumor efficacy was evaluated in BT-20 xenograft models.
Main Results:
- NHPI inhibited proliferation and induced G2/M phase arrest in cancer cells by downregulating cyclin B1 and cdc2.
- Apoptosis was induced via the mitochondrial pathway, and NHPI effectively inhibited both mTORC1 and mTORC2 signaling.
- In vivo, NHPI demonstrated tumor growth inhibition in xenografts with no significant toxicity, while suppressing mTOR signaling.
Conclusions:
- NHPI possesses antitumor activity linked to the inhibition of the mTOR signaling pathway.
- NHPI shows promise as a potential cancer therapeutic agent targeting the mTOR pathway, warranting further investigation.
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