Sangivamycin and its derivatives inhibit Haspin-Histone H3-survivin signaling and induce pancreatic cancer cell death

Abstract Current treatment options for patients with pancreatic cancer are suboptimal, resulting in a five year survival rate of about 9%. Difficulties with treatment are due to an immunosuppressive, fibrotic tumor microenvironment that prevents drugs from reaching tumor cells, but also to the limited efficacy of existing FDA-approved chemotherapeutic compounds. We here show that the nucleoside analog Sangivamycin and its closely-related compound Toyocamycin target PDA cell lines, and are significantly more efficient than Gemcitabine. Using KINOMEscan screening, we identified the kinase Haspin, which is overexpressed in PDA cell lines and human PDA samples, as a main target for both compounds. Inhibition of Haspin leads to a decrease in Histone H3 phosphorylation and prevents Histone H3 binding to survivin, thus providing mechanistic insight of how Sangivamycin targets cell proliferation, mitosis and induces apoptotic cell death. In orthotopically implanted tumors in mice, Sangivamycin was efficient in decreasing the growth of established tumors. In summary, we show that Sangivamycin and derivatives can be an efficient new option for treatment of PDA.

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The Bispidinone Derivative 3,7-Bis-[2-(S)-amino-3-(1H-indol-3-yl)-propionyl]-1,5-diphenyl-3,7-diazabicyclo[3.3.1]nonan-9-one Dihydrochloride Induces an Apoptosis-Mediated Cytotoxic Effect on Pancreatic Cancer Cells In Vitro

Molecules ◽

10.3390/molecules24030524 ◽

2019 ◽

Vol 24 (3) ◽

pp. 524 ◽

Cited By ~ 1

Author(s):

Melanie Predebon ◽

Danielle Bond ◽

Joshua Brzozowski ◽

Helen Jankowski ◽

Fiona Deane ◽

...

Keyword(s):

Pancreatic Cancer ◽

Flow Cytometry ◽

Cell Death ◽

Cell Lines ◽

Cytotoxic Effect ◽

Treatment Time ◽

Apoptotic Cell ◽

Apoptotic Cell Death ◽

Cytotoxic Agents ◽

Cell Counting

Pancreatic cancer (PC) is a complex, heterogeneous disease with a dismal prognosis. Current therapies have failed to improve survival outcomes, urging the need for discovery of novel targeted treatments. Bispidinone derivatives have yet to be investigated as cytotoxic agents against PC cells. The cytotoxic effect of four bispidinone derivatives (BisP1: 1,5-diphenyl-3,7-bis(2-hydroxyethyl)-3,7-diazabicyclo[3.3.1]nonan-9-one; BisP2: 3,7-bis-(2-(S)-amino-4-methylsulfanylbutyryl)-1,5-diphenyl-3,7-diazabicyclo[3.3.1]nonan-9-one dihydrochloride; BisP3: [2-{7-[2-(S)-tert-butoxycarbonylamino-3-(1H-indol-3-yl)-propionyl]-9-oxo-1,5-diphenyl-3,7-diazabicyclo[3.3.1]non-3-yl}-1-(S)-(1H-indol-3-ylmethyl)-2-oxoethyl]-carbamic acid tertbutyl ester; BisP4: 3,7-bis-[2-(S)-amino-3-(1H-indol-3-yl)-propionyl]-1,5-diphenyl-3,7-diazabicyclo[3.3.1]nonan-9-one dihydrochloride) was assessed against PC cell lines (MiaPaca-2, CFPAC-1 and BxPC-3). Cell viability was assessed using a Cell Counting Kit-8 (CCK-8) colorimetric assay, while apoptotic cell death was confirmed using fluorescence microscopy and flow cytometry. Initial viability screening revealed significant cytotoxic activity from BisP4 treatment (1 µM–100 µM) on all three cell lines, with IC50 values for MiaPaca-2, BxPC-3, and CFPAC-1 16.9 µM, 23.7 µM, and 36.3 µM, respectively. Cytotoxic treatment time-response (4 h, 24 h, and 48 h) revealed a 24 h treatment time was sufficient to produce a cytotoxic effect on all cell lines. Light microscopy evaluation (DAPI staining) of BisP4 treated MiaPaca-2 PC cells revealed dose-dependent characteristic apoptotic morphological changes. In addition, flow cytometry confirmed BisP4 induced apoptotic cell death induction of activated caspase-3/-7. The bispidinone derivative BisP4 induced an apoptosis-mediated cytotoxic effect on MiaPaca-2 cell lines and significant cytotoxicity on CFPAC-1 and BxPC-3 cell lines. Further investigations into the precise cellular mechanisms of action of this class of compounds are necessary for potential development into pre-clinical trials.

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Induction of apoptotic cell death by direct-current treatment in human leukemic cell lines

Journal of Cancer Research and Clinical Oncology ◽

10.1007/s004320050073 ◽

1997 ◽

Vol 123 (7) ◽

pp. 370-376 ◽

Cited By ~ 2

Author(s):

Masatsugu Kurokawa ◽

Hiroshi Sakagami ◽

Fumio Kokubu ◽

Hiromichi Noda ◽

Minoru Takeda ◽

...

Keyword(s):

Cell Death ◽

Cell Lines ◽

Direct Current ◽

Apoptotic Cell ◽

Leukemic Cell ◽

Current Treatment ◽

Apoptotic Cell Death ◽

Human Leukemic Cell ◽

Leukemic Cell Lines

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Effect of hyperthermia combined with gemcitabine on apoptotic cell death in cultured human pancreatic cancer cell lines

International Journal of Hyperthermia ◽

10.1080/02656730802657036 ◽

2009 ◽

Vol 25 (3) ◽

pp. 210-219 ◽

Cited By ~ 46

Author(s):

Satoko Adachi ◽

Satoshi Kokura ◽

Tetsuya Okayama ◽

Takeshi Ishikawa ◽

Tomohisa Takagi ◽

...

Keyword(s):

Pancreatic Cancer ◽

Cell Death ◽

Cell Lines ◽

Cancer Cell ◽

Pancreatic Cancer Cell ◽

Apoptotic Cell ◽

Cancer Cell Lines ◽

Apoptotic Cell Death ◽

Human Pancreatic Cancer ◽

Human Pancreatic Cancer Cell

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Induction of apoptotic cell death by direct-current treatment in human leukemic cell lines

Journal of Cancer Research and Clinical Oncology ◽

10.1007/bf01240119 ◽

1997 ◽

Vol 123 (7) ◽

pp. 370-376 ◽

Cited By ~ 4

Author(s):

Masatsugu Kurokawa ◽

Hiroshi Sakagami ◽

Fumio Kokubu ◽

Hiromichi Noda ◽

Minoru Takeda ◽

...

Keyword(s):

Cell Death ◽

Cell Lines ◽

Direct Current ◽

Apoptotic Cell ◽

Leukemic Cell ◽

Current Treatment ◽

Apoptotic Cell Death ◽

Human Leukemic Cell ◽

Leukemic Cell Lines

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Liver X receptor inhibition potentiates mitotane-induced adrenotoxicity in ACC

Endocrine Related Cancer ◽

10.1530/erc-20-0031 ◽

2020 ◽

Vol 27 (6) ◽

pp. 361-373

Author(s):

Kate M Warde ◽

Erik Schoenmakers ◽

Eduardo Ribes Martinez ◽

Yi Jan Lim ◽

Maeve Leonard ◽

...

Keyword(s):

Cell Death ◽

Cell Lines ◽

Cholesterol Ester ◽

Free Cholesterol ◽

Cholesterol Metabolism ◽

Apoptotic Cell ◽

Treatment Options ◽

Liver X Receptor ◽

Resistant Disease ◽

Cholesterol Storage

Adrenocortical carcinoma (ACC) is a rare aggressive malignancy with a poor outcome largely due to limited treatment options. Here, we propose a novel therapeutic approach through modulating intracellular free cholesterol via the liver X receptor alpha (LXRα) in combination with current first-line pharmacotherapy, mitotane. H295R and MUC-1 ACC cell lines were pretreated with LXRα inhibitors in combination with mitotane. In H295R, mitotane (20, 40 and 50 µM) induced dose-dependent cell death; however, in MUC-1, this only occurred at a supratherapeutic concentration (200 µM). LXRα inhibition potentiated mitotane-induced cytotoxicity in both cell lines. This was confirmed through use of the CompuSyn model which showed moderate pharmacological synergism and was indicative of apoptotic cell death via an increase in annexinV and cleaved-caspase 3 expression. Inhibition of LXRα was confirmed through downregulation of cholesterol efflux pumps ABCA1 and ABCG1; however, combination treatment with mitotane attenuated this effect. Intracellular free-cholesterol levels were associated with increased cytotoxicity in H295R (r2 = 0.5210) and MUC-1 (r2 = 0.9299) cells. While both cell lines exhibited similar levels of free cholesterol at baseline, H295R were cholesterol ester rich, whereas MUC-1 were cholesterol ester poor. We highlight the importance of LXRα mediated cholesterol metabolism in the management of ACC, drawing attention to its role in the therapeutics of mitotane sensitive tumours. We also demonstrate significant differences in cholesterol storage between mitotane sensitive and resistant disease.

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Influence of irradiation and pentoxifylline on histone H3 phosphorylation in human tumour cell lines

Cell Proliferation ◽

10.1046/j.1365-2184.2002.00224.x ◽

2002 ◽

Vol 35 (1) ◽

pp. 37-43 ◽

Cited By ~ 4

Author(s):

A. Binder ◽

L. Bohm

Keyword(s):

Cell Lines ◽

Tumour Cell ◽

Histone H3 ◽

Human Tumour ◽

H3 Phosphorylation ◽

Tumour Cell Lines ◽

Histone H3 Phosphorylation ◽

Human Tumour Cell Lines

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EGFR-independent activation of p38 MAPK and EGFR-dependent activation of ERK1/2 are required for ROS-induced renal cell death

AJP Renal Physiology ◽

10.1152/ajprenal.00132.2004 ◽

2004 ◽

Vol 287 (5) ◽

pp. F1049-F1058 ◽

Cited By ~ 61

Author(s):

Jing Dong ◽

Sampath Ramachandiran ◽

Kulbhushan Tikoo ◽

Zhe Jia ◽

Serrine S. Lau ◽

...

Keyword(s):

Cell Death ◽

P38 Mapk ◽

Egf Receptor ◽

Histone H3 ◽

Dominant Negative ◽

Mapk Activation ◽

Transfected Cells ◽

H3 Phosphorylation ◽

Hsp27 Phosphorylation ◽

Histone H3 Phosphorylation

2,3,5-Tris-(glutathion- S-yl)hydroquinone (TGHQ), a reactive metabolite of the nephrotoxicant hydroquinone, induces the ROS-dependent activation of MAPKs, followed by histone H3 phosphorylation and oncotic cell death in renal proximal tubule epithelial cells (LLC-PK1). Cell death and histone H3 phosphorylation are attenuated by pharmacological inhibition of p38 MAPK or ERK1/2 pathways. Because TGHQ, but not epidermal growth factor (EGF), induces histone H3 phosphorylation and cell death in LLC-PK1 cells, we hypothesized that there are differences in the mechanisms by which TGHQ and EGF induce activation of the EGF receptor (EGFR). We therefore compared the relative ability of TGHQ, H2O2, and EGF to activate EGFR and MAPKs and found that p38 MAPK activation is EGFR independent, whereas ERK1/2 activation occurs mainly through EGFR activation. TGHQ, H2O2, and EGF induce different EGFR tyrosine phosphorylation profiles that likely influence the subsequent differential kinetics of MAPK activation. We next transfected LLC-PK1 cells with a dominant negative p38 MAPK-expressing plasmid (pcDNA3-DNp38). TGHQ failed to induce phosphorylation of p38 MAPK and its substrate, MK-2, in pcDNA3-DNp38-transfected cells, indicating loss of function of p38 MAPK. In untransfected, pcDNA3 or pcDNA3-p38 (native)-transfected LLC-PK1 cells, Hsp27 was intensively phosphorylated after TGHQ treatment, whereas in pcDNA3-DNp38-transfected cells, TGHQ failed to induce Hsp27 phosphorylation. Thus EGFR-independent p38 MAPK and EGFR-dependent ERK1/2 activation by TGHQ lead to the activation of two downstream signaling factors, i.e., histone H3 and Hsp27 phosphorylation, which have in common the potential ability to remodel chromatin.

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