Engagement with tNOX (ENOX2) to Inhibit SIRT1 and Activate p53-Dependent and -Independent Apoptotic Pathways by Novel 4,11-Diaminoanthra[2,3-<i>b</i>]furan-5,10-diones in Hepatocellular Carcinoma Cells
Hepatocellular carcinoma (HCC) is the most frequent primary malignancy of the liver and is among the top three causes of cancer-associated death worldwide. However, the clinical use of chemotherapy for HCC has been limited by various challenges, emphasizing the urgent need for novel agents with impr...
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| Format: | Article |
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MDPI AG
2019-03-01
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| Series: | Cancers |
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| Online Access: | https://www.mdpi.com/2072-6694/11/3/420 |
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| author | Chia-Yang Lin Atikul Islam Claire J. Su Alexander S. Tikhomirov Andrey E. Shchekotikhin Show-Mei Chuang Pin Ju Chueh Yao Li Chen |
| author_facet | Chia-Yang Lin Atikul Islam Claire J. Su Alexander S. Tikhomirov Andrey E. Shchekotikhin Show-Mei Chuang Pin Ju Chueh Yao Li Chen |
| author_sort | Chia-Yang Lin |
| collection | DOAJ |
| description | Hepatocellular carcinoma (HCC) is the most frequent primary malignancy of the liver and is among the top three causes of cancer-associated death worldwide. However, the clinical use of chemotherapy for HCC has been limited by various challenges, emphasizing the urgent need for novel agents with improved anticancer properties. We recently synthesized and characterized a series of 4,11-diaminoanthra[2,3-<i>b</i>]furan-5,10-dione derivatives that exhibit potent apoptotic activity against an array of cancer cell lines, including variants with multidrug resistance. Their effect on liver cancer cells, however, was unknown. Here, we investigated three selected 4,11-diaminoanthra[2,3-<i>b</i>]furan-5,10-dione derivatives (compounds <b>1</b>–<b>3</b>) for their cytotoxicity and the underlying molecular mechanisms in wild-type or p53-deficient HCC cells. Cytotoxicity was determined by WST-1 assays and cell impedance measurements and apoptosis was analyzed by flow cytometry. The interaction between compounds and tumor-associated NADH oxidase (tNOX, ENOX2) was studied by cellular thermal shift assay (CETSA). We found that compound <b>1</b> and <b>2</b> induced significant cytotoxicity in both HepG2 and Hep3B lines. CETSA revealed that compounds <b>1</b> and <b>2</b> directly engaged with tNOX, leading to a decrease in the cellular NAD<sup>+</sup>/NADH ratio. This decreased the NAD<sup>+</sup>-dependent activity of Sirtuin 1 (SIRT1) deacetylase. In p53-wild-type HepG2 cells, p53 acetylation/activation was enhanced, possibly due to the reduction in SIRT1 activity, and apoptosis was observed. In p53-deficient Hep3B cells, the reduction in SIRT1 activity increased the acetylation of c-Myc, thereby reactivating the TRAIL pathway and, ultimately leading to apoptosis. These compounds thus trigger apoptosis in both cell types, but via different pathways. Taken together, our data show that derivatives <b>1</b> and <b>2</b> of 4,11-diaminoanthra[2,3-<i>b</i>]furan-5,10-diones engage with tNOX and inhibit its oxidase activity. This results in cytotoxicity via apoptosis through tNOX-SIRT1 axis to enhance the acetylation of p53 or c-Myc in HCC cells, depending on their p53 status. |
| first_indexed | 2024-03-12T11:15:13Z |
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| institution | Directory Open Access Journal |
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| language | English |
| last_indexed | 2024-03-12T11:15:13Z |
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| spelling | doaj.art-ee4435fd72c94cd2aa376a2da1c9f4ac2023-09-02T02:19:44ZengMDPI AGCancers2072-66942019-03-0111342010.3390/cancers11030420cancers11030420Engagement with tNOX (ENOX2) to Inhibit SIRT1 and Activate p53-Dependent and -Independent Apoptotic Pathways by Novel 4,11-Diaminoanthra[2,3-<i>b</i>]furan-5,10-diones in Hepatocellular Carcinoma CellsChia-Yang Lin0Atikul Islam1Claire J. Su2Alexander S. Tikhomirov3Andrey E. Shchekotikhin4Show-Mei Chuang5Pin Ju Chueh6Yao Li Chen7Institute of Biomedical Sciences, National Chung Hsing University, Taichung 40227, TaiwanInstitute of Biomedical Sciences, National Chung Hsing University, Taichung 40227, TaiwanInstitute of Biomedical Sciences, National Chung Hsing University, Taichung 40227, TaiwanGause Institute of New Antibiotics, 11B. Pirogovskaya Street, 119021 Moscow, RussiaGause Institute of New Antibiotics, 11B. Pirogovskaya Street, 119021 Moscow, RussiaInstitute of Biomedical Sciences, National Chung Hsing University, Taichung 40227, TaiwanInstitute of Biomedical Sciences, National Chung Hsing University, Taichung 40227, TaiwanDepartment of General Surgery, Changhua Christian Hospital, Changhua 50008, TaiwanHepatocellular carcinoma (HCC) is the most frequent primary malignancy of the liver and is among the top three causes of cancer-associated death worldwide. However, the clinical use of chemotherapy for HCC has been limited by various challenges, emphasizing the urgent need for novel agents with improved anticancer properties. We recently synthesized and characterized a series of 4,11-diaminoanthra[2,3-<i>b</i>]furan-5,10-dione derivatives that exhibit potent apoptotic activity against an array of cancer cell lines, including variants with multidrug resistance. Their effect on liver cancer cells, however, was unknown. Here, we investigated three selected 4,11-diaminoanthra[2,3-<i>b</i>]furan-5,10-dione derivatives (compounds <b>1</b>–<b>3</b>) for their cytotoxicity and the underlying molecular mechanisms in wild-type or p53-deficient HCC cells. Cytotoxicity was determined by WST-1 assays and cell impedance measurements and apoptosis was analyzed by flow cytometry. The interaction between compounds and tumor-associated NADH oxidase (tNOX, ENOX2) was studied by cellular thermal shift assay (CETSA). We found that compound <b>1</b> and <b>2</b> induced significant cytotoxicity in both HepG2 and Hep3B lines. CETSA revealed that compounds <b>1</b> and <b>2</b> directly engaged with tNOX, leading to a decrease in the cellular NAD<sup>+</sup>/NADH ratio. This decreased the NAD<sup>+</sup>-dependent activity of Sirtuin 1 (SIRT1) deacetylase. In p53-wild-type HepG2 cells, p53 acetylation/activation was enhanced, possibly due to the reduction in SIRT1 activity, and apoptosis was observed. In p53-deficient Hep3B cells, the reduction in SIRT1 activity increased the acetylation of c-Myc, thereby reactivating the TRAIL pathway and, ultimately leading to apoptosis. These compounds thus trigger apoptosis in both cell types, but via different pathways. Taken together, our data show that derivatives <b>1</b> and <b>2</b> of 4,11-diaminoanthra[2,3-<i>b</i>]furan-5,10-diones engage with tNOX and inhibit its oxidase activity. This results in cytotoxicity via apoptosis through tNOX-SIRT1 axis to enhance the acetylation of p53 or c-Myc in HCC cells, depending on their p53 status.https://www.mdpi.com/2072-6694/11/3/420apoptosisanthraquinone derivativeshepatocellular carcinomap53tumor-associated NADH oxidase (tNOX or ENOX2)sirtuin 1 (SIRT1)c-Mycprotein acetylation |
| spellingShingle | Chia-Yang Lin Atikul Islam Claire J. Su Alexander S. Tikhomirov Andrey E. Shchekotikhin Show-Mei Chuang Pin Ju Chueh Yao Li Chen Engagement with tNOX (ENOX2) to Inhibit SIRT1 and Activate p53-Dependent and -Independent Apoptotic Pathways by Novel 4,11-Diaminoanthra[2,3-<i>b</i>]furan-5,10-diones in Hepatocellular Carcinoma Cells Cancers apoptosis anthraquinone derivatives hepatocellular carcinoma p53 tumor-associated NADH oxidase (tNOX or ENOX2) sirtuin 1 (SIRT1) c-Myc protein acetylation |
| title | Engagement with tNOX (ENOX2) to Inhibit SIRT1 and Activate p53-Dependent and -Independent Apoptotic Pathways by Novel 4,11-Diaminoanthra[2,3-<i>b</i>]furan-5,10-diones in Hepatocellular Carcinoma Cells |
| title_full | Engagement with tNOX (ENOX2) to Inhibit SIRT1 and Activate p53-Dependent and -Independent Apoptotic Pathways by Novel 4,11-Diaminoanthra[2,3-<i>b</i>]furan-5,10-diones in Hepatocellular Carcinoma Cells |
| title_fullStr | Engagement with tNOX (ENOX2) to Inhibit SIRT1 and Activate p53-Dependent and -Independent Apoptotic Pathways by Novel 4,11-Diaminoanthra[2,3-<i>b</i>]furan-5,10-diones in Hepatocellular Carcinoma Cells |
| title_full_unstemmed | Engagement with tNOX (ENOX2) to Inhibit SIRT1 and Activate p53-Dependent and -Independent Apoptotic Pathways by Novel 4,11-Diaminoanthra[2,3-<i>b</i>]furan-5,10-diones in Hepatocellular Carcinoma Cells |
| title_short | Engagement with tNOX (ENOX2) to Inhibit SIRT1 and Activate p53-Dependent and -Independent Apoptotic Pathways by Novel 4,11-Diaminoanthra[2,3-<i>b</i>]furan-5,10-diones in Hepatocellular Carcinoma Cells |
| title_sort | engagement with tnox enox2 to inhibit sirt1 and activate p53 dependent and independent apoptotic pathways by novel 4 11 diaminoanthra 2 3 i b i furan 5 10 diones in hepatocellular carcinoma cells |
| topic | apoptosis anthraquinone derivatives hepatocellular carcinoma p53 tumor-associated NADH oxidase (tNOX or ENOX2) sirtuin 1 (SIRT1) c-Myc protein acetylation |
| url | https://www.mdpi.com/2072-6694/11/3/420 |
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