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. 2019 Mar;19(3):1654-1664.
doi: 10.3892/mmr.2018.9785. Epub 2018 Dec 21.

Novel 1,4‑naphthoquinone derivatives induce reactive oxygen species‑mediated apoptosis in liver cancer cells

Yue Wang  1 , Ying-Hua Luo  2 , Xian-Ji Piao  3 , Gui-Nan Shen  1 , Ling-Qi Meng  1 , Yi Zhang  1 , Jia-Ru Wang  1 , Jin-Qian Li  1 , Hao Wang  1 , Wan-Ting Xu  1 , Yang Liu  1 , Yu Zhang  1 , Tong Zhang  1 , Shi-Nong Wang  1 , Hu-Nan Sun  1 , Ying-Hao Han  1 , Mei-Hua Jin  1 , Yan-Qing Zang  4 , Dong-Jie Zhang  4 , Cheng-Hao Jin  1
Affiliations

Novel 1,4‑naphthoquinone derivatives induce reactive oxygen species‑mediated apoptosis in liver cancer cells

Yue Wang et al. Mol Med Rep. 2019 Mar.

Abstract

Derivatives of 1,4‑naphthoquinone have excellent anti‑cancer effects, but their use has been greatly limited due to their serious side effects. To develop compounds with decreased side effects and improved anti‑cancer activity, two novel types of 1,4‑naphthoquinone derivatives, 2,3‑dihydro‑2,3‑epoxy‑2‑propylsulfonyl‑5,8‑dimethoxy‑1,4‑naphthoquinone (EPDMNQ) and 2,3‑dihydro‑2,3‑epoxy‑2‑nonylsulfonyl‑5,8‑dimethoxy‑1,4‑naphthoquinone (ENDMNQ) were synthesized and their anti‑tumor activities were investigated. The effects of EPDMNQ and ENDMNQ on cell viability, apoptosis and accumulation of reactive oxygen species (ROS) in liver cancer cells were determined by MTT cell viability assay and flow cytometry. The expression levels of mitochondrial, mitogen activated protein kinase (MAPK) and signal transducer and activator of transcription 3 (STAT3) signaling pathway‑associated proteins in Hep3B liver cancer cells were analyzed by western blot analysis. The results demonstrated that EPDMNQ and ENDMNQ inhibited the proliferation of liver cancer Hep3B, HepG2, and Huh7 cell lines but not that of normal liver L‑02, normal lung IMR‑90 and stomach GES‑1 cell lines. The number of apoptotic cells and ROS levels were significantly increased following treatment with EPDMNQ and ENDMNQ, and these effects were blocked by the ROS inhibitor N‑acetyl‑L‑cysteine (NAC) in Hep3B cells. EPDMNQ and ENDMNQ induced apoptosis by upregulating the protein expression of p38 MAPK and c‑Jun N‑terminal kinase and downregulating extracellular signal‑regulated kinase and STAT3; these effects were inhibited by NAC. The results of the present study demonstrated that EPDMNQ and ENDMNQ induced apoptosis through ROS‑modulated MAPK and STAT3 signaling pathways in Hep3B cells. Therefore, these novel 1,4‑naphthoquinone derivatives may be useful as anticancer agents for the treatment of liver cancer.

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Figures

Figure 1.
Figure 1.
Synthesis of 1,4-naphthoquinone derivatives EPDMNQ and ENDMNQ. (A) Process of synthetic EPDMNQ and ENDMNQ generation. (B) Structural formulas of EPDMNQ and ENDMNQ. EPDMNQ, 2,3-dihydro-2,3-epoxy-2-propylsulfonyl-5,8-dimethoxy-1,4-naphthoquinone; ENDMNQ, 2,3-dihydro-2,3-epoxy-2-nonylsulfonyl-5,8-dimethoxy-1,4-naphthoquinone.
Figure 2.
Figure 2.
Effects of EPDMNQ and ENDMNQ on the viabilities of liver cancer and normal cells. (A) Hep3B, HepG2 and Huh7 cells were treated with different concentrations (1, 3, 10, 30 or 100 µmol/l) of 5-FU, EPDMNQ or ENDMNQ for 24 h. Cell viability was determined by MTT assay. (B) Normal liver L-02, normal lung IMR-90 and normal stomach GES-1 cell line viabilities. Data are expressed as the percentage of viable cells. ***P<0.001 vs. 5-FU group. EPDMNQ, 2,3-dihydro-2,3-epoxy-2-propylsulfonyl-5,8-dimethoxy-1,4-naphthoquinone; ENDMNQ, 2,3-dihydro-2,3-epoxy-2-nonylsulfonyl-5,8-dimethoxy-1,4-naphthoquinone; 5-FU, 5-fluorouracil.
Figure 3.
Figure 3.
EPDMNQ and ENDMNQ induce apoptosis in hepatocellular carcinoma cells. (A) Hep3B cells were treated with 4 µmol/l 5-FU, EPDMNQ or ENDMNQ for different time intervals (3, 6, 12 or 24 h) and stained with Annexin V-FITC/PI. Images represent fluorescence microscopic images (original magnifications, ×400). (B) Quantification of fluorescence intensities from A. (C) Apoptosis distribution was determined by flow cytometry following treatment with 4 µmol/l 5-FU, EPDMNQ or ENDMNQ for different time intervals (3, 6, 12 or 24 h). (D) Quantification of flow cytometry from C. Data are expressed as the percentage of viable cells. ***P<0.001 vs. 5-FU group. EPDMNQ, 2,3-dihydro-2,3-epoxy-2-propylsulfonyl-5,8-dimethoxy-1,4-naphthoquinone; ENDMNQ, 2,3-dihydro-2,3-epoxy-2-nonylsulfonyl-5,8-dimethoxy-1,4-naphthoquinone; 5-FU, 5-fluorouracil; FITC, fluorescein isothiocyanate; PI, propidium iodide.
Figure 4.
Figure 4.
EPDMNQ and ENDMNQ regulate mitochondrial, MAPK and STAT3 signaling pathways-associated proteins in Hep3B cells. (A) Hep3B cells were treated with EPDMNQ or ENDMNQ for different time intervals (3, 6, 12 or 24 h). The expression levels of mitochondrial pathway-associated proteins were analyzed by western blot analysis. (B) The expression levels of MAPK and STAT3 pathway-associated proteins were analyzed by western blot analysis. α-tubulin was used as the internal control. Band intensity was quantified using the Image J software. **P<0.01 and ***P<0.001 vs. untreated control group. EPDMNQ, 2,3-dihydro-2,3-epoxy-2-propylsulfonyl-5,8-dimethoxy-1,4-naphthoquinone; ENDMNQ, 2,3-dihydro-2,3-epoxy-2-nonylsulfonyl-5,8-dimethoxy-1,4-naphthoquinone; Bcl-2, B-cell lymphoma 2; Bax, Bcl-2-associated X protein; cle, cleaved; PARP, poly (adenosine 5-diphosphate-ribose) polymerase; p, phosphorylated; p38, p38 mitogen-activated protein kinase; JNK, c-Jun N-terminal kinase; ERK, extracellular signal regulated kinase; STAT3, signal transducer and activator of transcription 3.
Figure 5.
Figure 5.
EPDMNQ and ENDMNQ-induced apoptosis is restored by ROS inhibition in Hep3B cells. (A and B) Intracellular ROS generation induced by EPDMNQ or ENDMNQ was measured in Hep3B cells following staining with 2′,7′-dichlorodihydrofluorescein diacetate and detected by flow cytometry. (C and D) Hep3B cells were treated with EPDMNQ or ENDMNQ for 24 h, and mitochondrial membrane potential depolarization value was detected by flow cytometry following JC-1 staining. (E) Hep3B cells were treated with NAC for 30 min and then incubated with EPDMNQ for 24 h. Cell apoptosis was determined by flow cytometry. (F) Apoptotic cell numbers were detected following ENDMNQ treatment by flow cytometry. Data are expressed as the apoptotic cell numbers. **P<0.01 and ***P<0.001 vs. untreated control group. EPDMNQ, 2,3-dihydro-2,3-epoxy-2-propylsulfonyl-5,8-dimethoxy-1,4-naphthoquinone; ENDMNQ, 2,3-dihydro-2,3-epoxy-2-nonylsulfonyl-5,8-dimethoxy-1,4-naphthoquinone; ROS, reactive oxygen species; NAC, N-acetyl-L-cysteine.
Figure 6.
Figure 6.
Roles of mitochondrial, mitogen-activated kinase and STAT3 signaling pathways in the apoptosis of EPDMNQ- and ENDMNQ-treated Hep3B cells. Detection of p-ERK, p-JNK, p-p38, p-STAT3, Bax, Bcl-2, cle-PARP and cle-caspase-3 protein expression in (A) EPDMNQ- and (B) ENDMNQ-treated Hep3B cells by western blot analysis. ***P<0.001 vs. untreated control groupl. EPDMNQ, 2,3-dihydro-2,3-epoxy-2-propylsulfonyl-5,8-dimethoxy-1,4-naphthoquinone; ENDMNQ, 2,3-dihydro-2,3-epoxy-2-nonylsulfonyl-5,8-dimethoxy-1,4-naphthoquinone; Bcl-2, B-cell lymphoma 2; Bax, Bcl-2-associated X protein; cle, cleaved; PARP, poly (adenosine 5-diphosphate-ribose) polymerase; p, phosphorylated; p38, p38 mitogen-activated protein kinase; JNK, c-Jun N-terminal kinase; ERK, extracellular signal regulated kinase; STAT3, signal transducer and activator of transcription 3.
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References

    1. Zhang ZQ, Meng H, Wang N, Liang LN, Liu LN, Lu SM, Luan Y. Serum microRNA 143 and microRNA 215 as potential biomarkers for the diagnosis of chronic hepatitis and hepatocellular carcinoma. Diagn Pathol. 2014;9:135. doi: 10.1186/1746-1596-9-135. - DOI - PMC - PubMed
    1. Sellam F, Harir N, Khaled MB, Mrabent NM, Salah R, Benchouk A, Diaf M. Delayed diagnosis of pancreatic cancer reported as more common in a population of North African young adults. J Gastrointest Oncol. 2015;6:505–510. - PMC - PubMed
    1. Bruix J, Reig M, Sherman M. Evidence-based diagnosis, staging, and treatment of patients with hepatocellular carcinoma. Gastroenterology. 2016;150:835–853. doi: 10.1053/j.gastro.2015.12.041. - DOI - PubMed
    1. Li Q, Zhu LZ, Yang RJ, Zhu X. Cytotoxic activity of anticancer drugs on hepatocellular carcinoma cells in hypoxic-hyponutritional culture. Int Surg. 2014;99:745–752. doi: 10.9738/INTSURG-D-14-00073.1. - DOI - PMC - PubMed
    1. Loureiro R, Magalhães-Novais S, Mesquita KA, Baldeiras I, Sousa IS, Tavares LC, Barbosa IA, Oliveira PJ, Vega-Naredo I. Melatonin antiproliferative effects require active mitochondrial function in embryonal carcinoma cells. Oncotarget. 2015;6:17081–17096. doi: 10.18632/oncotarget.4012. - DOI - PMC - PubMed

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