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Article ; Online: Regulation of colorectal cancer cell epithelial to mesenchymal transition by the renin angiotensin system.

Nguyen, Linh / Ager, Eleanor I / Neo, Jaclyn / Christophi, Christopher

Journal of gastroenterology and hepatology

2016 Volume 31, Issue 10, Page(s) 1773–1782

Abstract: Background and aim: Epithelial to mesenchymal transition (EMT) is implicated in tumor progression. We aimed to determine if the renin angiotensin system has a role in colorectal cancer (CRC) cell EMT.: Methods: Human CRC cell lines DLD-1 and LIM2405 ... ...

Abstract	Background and aim: Epithelial to mesenchymal transition (EMT) is implicated in tumor progression. We aimed to determine if the renin angiotensin system has a role in colorectal cancer (CRC) cell EMT. Methods: Human CRC cell lines DLD-1 and LIM2405 were used in wound scratch migration assays where they were treated with renin angiotensin system peptide ANG II alone or with blockers of ANG II type 1 or 2 receptors (AT1R and AT2R). Levels of epithelial (E-cadherin), mesenchymal (ZEB1, Vimentin) markers, inducible nitric oxide synthase (iNOS), and MMP9 were determined by flow cytometry. Mice bearing CRC liver metastases and treated with blockers for AT1R or AT2R were examined for ZEB1 and iNOS by immunohistochemistry. Results: ANG II increased in-vitro CRC cell migration in both cell lines, this was inhibited by AT1R (IRB) or AT2R blockade (PD123319). DLD-1 cells treated with AT1R blocker resulted in increased E-cadherin, reduced ZEB1, and Vimentin expression compared with ANG II-treated cells. Treatment with AT2R blocker decreased E-cadherin, no change in ZEB1 or Vimentin expression. AT1R blockade increased iNOS and decreased MMP9 expression in DLD-1 and LIM2405 cells. AT2R blockade decreased iNOS and MMP9 expression in both cell lines. In vivo, ZEB1 staining was higher in ANG II-treated animals compared with control and AT1R blockade treated animals, while activation of the AT2R led to an increase in iNOS compared with control and AT1R blockade. Conclusions: ANG II-induced migration of CRC cells via both AT1 and AT2 receptors; the AT1R-mediated effects were associated with changes typical of EMT.
MeSH term(s)	Angiotensin II/pharmacology ; Angiotensin II Type 1 Receptor Blockers/pharmacology ; Animals ; Cell Movement/drug effects ; Colorectal Neoplasms/metabolism ; Colorectal Neoplasms/pathology ; Colorectal Neoplasms/physiopathology ; Epithelial-Mesenchymal Transition/physiology ; Humans ; Liver Neoplasms/pathology ; Liver Neoplasms/secondary ; Matrix Metalloproteinase 9/biosynthesis ; Mice, Inbred CBA ; Neoplasm Proteins/metabolism ; Neoplasm Transplantation ; Nitric Oxide Synthase Type II/metabolism ; Receptor, Angiotensin, Type 1/metabolism ; Receptor, Angiotensin, Type 2/metabolism ; Renin-Angiotensin System/physiology ; Tumor Cells, Cultured
Chemical Substances	Angiotensin II Type 1 Receptor Blockers ; Neoplasm Proteins ; Receptor, Angiotensin, Type 1 ; Receptor, Angiotensin, Type 2 ; Angiotensin II (11128-99-7) ; NOS2 protein, human (EC 1.14.13.39) ; Nitric Oxide Synthase Type II (EC 1.14.13.39) ; MMP9 protein, human (EC 3.4.24.35) ; Matrix Metalloproteinase 9 (EC 3.4.24.35)
Language	English
Publishing date	2016-10
Publishing country	Australia
Document type	Journal Article
ZDB-ID	632882-9
ISSN	1440-1746 ; 0815-9319
ISSN (online)	1440-1746
ISSN	0815-9319
DOI	10.1111/jgh.13307
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Article ; Online: Bimodal role of Kupffer cells during colorectal cancer liver metastasis.

Wen, Shu Wen / Ager, Eleanor I / Christophi, Christopher

Cancer biology & therapy

2013 Volume 14, Issue 7, Page(s) 606–613

Abstract: Kupffer cells (KCs) are resident liver macrophages that play a crucial role in liver homeostasis and in the pathogenesis of liver disease. Evidence suggests KCs have both stimulatory and inhibitory functions during tumor development but the extent of ... ...

Abstract	Kupffer cells (KCs) are resident liver macrophages that play a crucial role in liver homeostasis and in the pathogenesis of liver disease. Evidence suggests KCs have both stimulatory and inhibitory functions during tumor development but the extent of these functions remains to be defined. Using KC depletion studies in an orthotopic murine model of colorectal cancer (CRC) liver metastases we demonstrated the bimodal role of KCs in determining tumor growth. KC depletion with gadolinium chloride before tumor induction was associated with an increased tumor burden during the exponential growth phase. In contrast, KC depletion at the late stage of tumor growth (day 18) decreased liver tumor load compared with non-depleted animals. This suggests KCs exhibit an early inhibitory and a later stimulatory effect. These two opposing functions were associated with changes in iNOS and VEGF expression as well as T-cell infiltration. KC depletion at day 18 increased numbers of CD3 (+) T cells and iNOS-expressing infiltrating cells in the tumor, but decreased the number of VEGF-expressing infiltrating cells. These alterations may be responsible for the observed reduction in tumor burden following depletion of pro-tumor KCs at the late stage of metastatic growth. Taken together, our results indicate that the bimodal role of KC activity in liver tumors may provide the key to timing immunomodulatory intervention for the treatment of CRC liver metastases.
MeSH term(s)	Animals ; Colorectal Neoplasms/metabolism ; Colorectal Neoplasms/pathology ; Humans ; Kupffer Cells/metabolism ; Kupffer Cells/pathology ; Liver Neoplasms, Experimental/metabolism ; Liver Neoplasms, Experimental/pathology ; Liver Neoplasms, Experimental/secondary ; Mice ; Neoplasm Metastasis
Language	English
Publishing date	2013-05-10
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2146305-0
ISSN	1555-8576 ; 1538-4047
ISSN (online)	1555-8576
ISSN	1538-4047
DOI	10.4161/cbt.24593
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Article ; Online: Mechanism of action of the third generation benzopyrans and evaluation of their broad anti-cancer activity in vitro and in vivo.

Stevenson, Alexander J / Ager, Eleanor I / Proctor, Martina A / Škalamera, Dubravka / Heaton, Andrew / Brown, David / Gabrielli, Brian G

Scientific reports

2018 Volume 8, Issue 1, Page(s) 5144

Abstract: Successive rounds of chemical modification in three generations of benzopyran molecules have shown to select for different mechanisms of actions and progressive increases in anti-cancer activity. In this study, we investigated the mechanism of action of ... ...

Abstract	Successive rounds of chemical modification in three generations of benzopyran molecules have shown to select for different mechanisms of actions and progressive increases in anti-cancer activity. In this study, we investigated the mechanism of action of the third-generation benzopyran compounds, TRX-E-002-1 and TRX-E-009-1. High-content screening of a panel of 240 cancer cell lines treated with TRX-E-009-1 demonstrated it has broad anti-cancer potential. Within this screen, melanoma cell lines showed a range of sensitivities and subsequently a second independent panel of 21 melanoma 3D spheroid lines were assessed for their responses to both TRX-E-002-1 and TRX-E-009-1 compounds. Time-lapse microscopy illustrated both of these compounds caused mitotic delays in treated cells, resulting in either mitotic slippage or apoptosis. This finding along with immunostaining, in vitro polymerization assays, and animal experiments in both athymic and immunocompetent mice, demonstrates that these third-generation benzopyran compounds are potent tubulin polymerization inhibitors in vitro and in vivo, and this is the molecular basis of their anti-cancer activity in melanoma. These findings indicate these BP compounds may offer a novel anti-microtubule strategy for cancer intervention and provides the basis for further investigation into biomarkers of clinical sensitivity.
MeSH term(s)	Animals ; Benzopyrans/chemistry ; Benzopyrans/pharmacology ; Cell Line, Tumor ; Flavonoids/chemistry ; Flavonoids/pharmacology ; Humans ; Melanoma, Experimental/drug therapy ; Melanoma, Experimental/metabolism ; Melanoma, Experimental/pathology ; Mice ; Mice, Nude ; Tubulin Modulators/chemistry ; Tubulin Modulators/pharmacology ; Xenograft Model Antitumor Assays
Chemical Substances	4-(para-hydroxyphenyl)-7,4'-dihydroxy-3',5'-dimethoxy-8-methylisoflavan ; Benzopyrans ; Flavonoids ; Tubulin Modulators
Language	English
Publishing date	2018-03-23
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/s41598-018-22882-w
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Article ; Online: Mechanism of action of the third generation benzopyrans and evaluation of their broad anti-cancer activity in vitro and in vivo

Alexander J. Stevenson / Eleanor I. Ager / Martina A. Proctor / Dubravka Škalamera / Andrew Heaton / David Brown / Brian G. Gabrielli

Scientific Reports, Vol 8, Iss 1, Pp 1-

2018 Volume 11

Abstract: Abstract Successive rounds of chemical modification in three generations of benzopyran molecules have shown to select for different mechanisms of actions and progressive increases in anti-cancer activity. In this study, we investigated the mechanism of ... ...

Abstract	Abstract Successive rounds of chemical modification in three generations of benzopyran molecules have shown to select for different mechanisms of actions and progressive increases in anti-cancer activity. In this study, we investigated the mechanism of action of the third-generation benzopyran compounds, TRX-E-002-1 and TRX-E-009-1. High-content screening of a panel of 240 cancer cell lines treated with TRX-E-009-1 demonstrated it has broad anti-cancer potential. Within this screen, melanoma cell lines showed a range of sensitivities and subsequently a second independent panel of 21 melanoma 3D spheroid lines were assessed for their responses to both TRX-E-002-1 and TRX-E-009-1 compounds. Time-lapse microscopy illustrated both of these compounds caused mitotic delays in treated cells, resulting in either mitotic slippage or apoptosis. This finding along with immunostaining, in vitro polymerization assays, and animal experiments in both athymic and immunocompetent mice, demonstrates that these third-generation benzopyran compounds are potent tubulin polymerization inhibitors in vitro and in vivo, and this is the molecular basis of their anti-cancer activity in melanoma. These findings indicate these BP compounds may offer a novel anti-microtubule strategy for cancer intervention and provides the basis for further investigation into biomarkers of clinical sensitivity.
Keywords	Medicine ; R ; Science ; Q
Language	English
Publishing date	2018-03-01T00:00:00Z
Publisher	Nature Publishing Group
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: The renin angiotensin system regulates Kupffer cells in colorectal liver metastases.

Wen, Shu Wen / Ager, Eleanor I / Neo, Jaclyn / Christophi, Christopher

Cancer biology & therapy

2013 Volume 14, Issue 8, Page(s) 720–727

Abstract: Blockade of the renin angiotensin system (RAS) can inhibit tumor growth and this may be mediated via undefined immunomodulatory actions. This study investigated the effects of RAS blockade on liver macrophages (Kupffer cells; KCs) in an orthotopic murine ...

Abstract	Blockade of the renin angiotensin system (RAS) can inhibit tumor growth and this may be mediated via undefined immunomodulatory actions. This study investigated the effects of RAS blockade on liver macrophages (Kupffer cells; KCs) in an orthotopic murine model of colorectal cancer (CRC) liver metastases. Here we showed that pharmacological targeting of the RAS [ANG II (31.25 µg/kg/h i.p.), ANG-(1-7) (24 µg/kg/h i.p.) or the ACE inhibitor; captopril (750 mg/kg/d i.p.)] altered endogenous KC numbers in the tumor-bearing liver throughout metastatic growth. Captopril, and to a lesser extent ANG-(1-7), increased KC numbers in the liver but not tumor. KCs were found to express the key RAS components: ACE and AT1R. Treatment with captopril and ANG II increased the number of AT1R-expressing KCs, although total KC numbers were not affected by ANG II. Captopril (0.1 µM) also increased macrophage invasion in vitro. Additionally, captopril was administered with KC depletion before tumor induction (day 0) or at established metastatic growth (day 18) using gadolinium chloride (GdCl 3; 20 mg/kg). Livers were collected at day 21 and quantitative stereology used as a measure of tumor burden. Captopril reduced growth of CRC liver metastases. However, when captopril was combined with early KC depletion (day 0) tumor growth was significantly increased compared with captopril alone. In contrast, late KC depletion (day 18) failed to influence the anti-tumor effects of captopril. The result of these studies suggests that manipulation of the RAS can alter KC numbers and may subsequently influence progression of CRC liver metastases.
MeSH term(s)	Angiotensin I/pharmacology ; Angiotensin II/pharmacology ; Angiotensin-Converting Enzyme Inhibitors/pharmacology ; Animals ; Captopril/pharmacology ; Colorectal Neoplasms/drug therapy ; Colorectal Neoplasms/metabolism ; Colorectal Neoplasms/pathology ; Disease Models, Animal ; Gadolinium/pharmacology ; Kupffer Cells/drug effects ; Kupffer Cells/pathology ; Liver Neoplasms/drug therapy ; Liver Neoplasms/metabolism ; Liver Neoplasms/secondary ; Macrophages/drug effects ; Macrophages/pathology ; Male ; Mice ; Mice, Inbred CBA ; Peptide Fragments/pharmacology ; Renin-Angiotensin System/drug effects ; Renin-Angiotensin System/physiology
Chemical Substances	Angiotensin-Converting Enzyme Inhibitors ; Peptide Fragments ; Angiotensin II (11128-99-7) ; Angiotensin I (9041-90-1) ; Captopril (9G64RSX1XD) ; Gadolinium (AU0V1LM3JT) ; angiotensin I (1-7) (IJ3FUK8MOF) ; gadolinium chloride (P7082WY76D)
Language	English
Publishing date	2013-06-17
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2146305-0
ISSN	1555-8576 ; 1538-4047
ISSN (online)	1555-8576
ISSN	1538-4047
DOI	10.4161/cbt.25092
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Article ; Online: Correction: Targeting the angiotensin II type 2 receptor (AT2R) in colorectal liver metastases.

Ager, Eleanor I / Chong, Way W / Wen, Shu-Wen / Christophi, Christopher

Cancer cell international

2010 Volume 10, Page(s) 37

Language	English
Publishing date	2010-10-21
Publishing country	England
Document type	Journal Article ; Published Erratum
ISSN	1475-2867
ISSN (online)	1475-2867
DOI	10.1186/1475-2867-10-37
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Article ; Online: Liver regeneration and tumour stimulation: implications of the renin-angiotensin system.

Koh, Shir Lin / Ager, Eleanor I / Christophi, Christopher

Liver international : official journal of the International Association for the Study of the Liver

2010 Volume 30, Issue 10, Page(s) 1414–1426

Abstract: Liver resection is the most effective treatment for primary liver tumours and metastasis to the liver, and remains the only potentially long-term curative therapy for patients with colorectal cancer (CRC) liver metastases. Nevertheless, there is a ... ...

Abstract	Liver resection is the most effective treatment for primary liver tumours and metastasis to the liver, and remains the only potentially long-term curative therapy for patients with colorectal cancer (CRC) liver metastases. Nevertheless, there is a significant incidence of tumour recurrence following liver resection. Cellular and molecular changes resulting from resection and the subsequent liver regeneration process may influence the kinetics of tumour growth, contributing to recurrence. Although commonly associated with the systemic homeostasis of blood pressure, fluid and electrolyte, the renin-angiotensin system (RAS) has recently been shown to play a role in regulating cell proliferation, apoptosis and angiogenesis in local organs as well as in malignancies. An electronic search of the English literature on the role of the RAS in liver regeneration and tumourigenesis was performed using PubMed, with additional relevant articles sourced from reference lists. Studies have shown that the blockade of the RAS pathway stimulates liver regeneration and inhibits tumour progression. An understanding of the role of RAS in liver regeneration and tumourigenesis may enable alternative strategies to improve patient outcome and survival after liver resection. This review will discuss the role of the RAS in liver regeneration and in tumour recurrence post-liver resection. The potential of the RAS as a novel therapeutic target for CRC liver metastases patients undergoing liver resection will be outlined.
MeSH term(s)	Angiotensin II Type 1 Receptor Blockers/therapeutic use ; Angiotensin-Converting Enzyme Inhibitors/therapeutic use ; Animals ; Antineoplastic Agents/therapeutic use ; Cell Proliferation/drug effects ; Chemotherapy, Adjuvant ; Hepatectomy/adverse effects ; Humans ; Liver/drug effects ; Liver/metabolism ; Liver/pathology ; Liver/surgery ; Liver Neoplasms/drug therapy ; Liver Neoplasms/metabolism ; Liver Neoplasms/pathology ; Liver Neoplasms/surgery ; Liver Regeneration/drug effects ; Neoplasm Recurrence, Local/etiology ; Neoplasm Recurrence, Local/metabolism ; Neoplasm Recurrence, Local/pathology ; Neoplasm Recurrence, Local/prevention & control ; Renin-Angiotensin System/drug effects ; Treatment Outcome
Chemical Substances	Angiotensin II Type 1 Receptor Blockers ; Angiotensin-Converting Enzyme Inhibitors ; Antineoplastic Agents
Language	English
Publishing date	2010-11
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	2102783-3
ISSN	1478-3231 ; 1478-3223
ISSN (online)	1478-3231
ISSN	1478-3223
DOI	10.1111/j.1478-3231.2010.02306.x
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Article ; Online: The renin-angiotensin system and malignancy.

Ager, Eleanor I / Neo, Jaclyn / Christophi, Christopher

Carcinogenesis

2008 Volume 29, Issue 9, Page(s) 1675–1684

Abstract: The renin-angiotensin system (RAS) is usually associated with its systemic action on cardiovascular homoeostasis. However, recent studies suggest that at a local tissue level, the RAS influences tumour growth. The potential of the RAS as a target for ... ...

Abstract	The renin-angiotensin system (RAS) is usually associated with its systemic action on cardiovascular homoeostasis. However, recent studies suggest that at a local tissue level, the RAS influences tumour growth. The potential of the RAS as a target for cancer treatment and the suggested underlying mechanisms of its paracrine effects are reviewed here. These include modulation of angiogenesis, cellular proliferation, immune responses and extracellular matrix formation. Knowledge of the RAS has increased dramatically in recent years with the discovery of new enzymes, peptides and feedback mechanisms. The local RAS appears to influence tumour growth and metastases and there is evidence of tissue- and tumour-specific differences. Recent experimental studies provide strong evidence that drugs that inhibit the RAS have the potential to reduce cancer risk or retard tumour growth and metastases. Manipulation of the RAS may, therefore, provide a safe and inexpensive anticancer strategy.
MeSH term(s)	Angiotensin-Converting Enzyme Inhibitors/therapeutic use ; Animals ; Humans ; Neoplasms/metabolism ; Neoplasms/pathology ; Renin-Angiotensin System/physiology
Chemical Substances	Angiotensin-Converting Enzyme Inhibitors
Language	English
Publishing date	2008-09
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	603134-1
ISSN	1460-2180 ; 0143-3334
ISSN (online)	1460-2180
ISSN	0143-3334
DOI	10.1093/carcin/bgn171
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Article ; Online: Targeting the angiotensin II type 2 receptor (AT2R) in colorectal liver metastases.

Ager, Eleanor I / Chong, Way W / Wen, Shu-Wen / Christophi, Christopher

Cancer cell international

2010 Volume 10, Page(s) 19

Abstract: Background: Blockade of the angiotensin (ANG) II type 1 receptor (AT1R) inhibits tumour growth in several cancers, including colorectal cancer (CRC) liver metastases. While AT1R blockade has been extensively studied, the potential of targeting the ... ...

Abstract	Background: Blockade of the angiotensin (ANG) II type 1 receptor (AT1R) inhibits tumour growth in several cancers, including colorectal cancer (CRC) liver metastases. While AT1R blockade has been extensively studied, the potential of targeting the antagonistically acting AT2R in cancer has not been investigated. This study examined the effect of AT2R activation with the agonist CGP42112A in a mouse model of CRC liver metastases. Results: In vitro, mouse CRC cell (MoCR) proliferation was inhibited by treatment with CGP42112A in a dose dependent manner while apoptosis was increased. Immunofluorescent staining for key signalling and secondary messengers, PLA2 and iNOS, were also increased by CGP42112A treatment in vitro. Immunohistochemical staining for proliferation (PCNA) and the apoptosis (active caspase 3) markers confirmed a CGP42112A-associated inhibition of proliferation and induction of apoptosis of mouse CRC cells (MoCR) in vivo. However, angiogenesis and vascular endothelial growth factor (VEGF) appeared to be increased by CGP42112A treatment in vivo. This increase in VEGF secretion by MoCRs was confirmed in vitro. Despite this apparent pro-angiogenic effect, a syngenic orthotopic mouse model of CRC liver metastases showed a reduction in liver to body weight ratio, an indication of tumour burden, following CGP42112A treatment compared to untreated controls. Conclusions: These results suggest that AT2R activation might provide a novel target to inhibit tumour growth. Its potential to stimulate angiogenesis could be compensated by combination with anti-angiogenic agents.
Language	English
Publishing date	2010-06-28
Publishing country	England
Document type	Journal Article
ISSN	1475-2867
ISSN (online)	1475-2867
DOI	10.1186/1475-2867-10-19
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Article ; Online: Expression and protein localisation of IGF2 in the marsupial placenta.

Ager, Eleanor I / Pask, Andrew J / Shaw, Geoff / Renfree, Marilyn B

BMC developmental biology

2008 Volume 8, Page(s) 17

Abstract: Background: In eutherian mammals, genomic imprinting is critical for normal placentation and embryo survival. Insulin-like growth factor 2 (IGF2) is imprinted in the placenta of both eutherians and marsupials, but its function, or that of any imprinted ... ...

Abstract	Background: In eutherian mammals, genomic imprinting is critical for normal placentation and embryo survival. Insulin-like growth factor 2 (IGF2) is imprinted in the placenta of both eutherians and marsupials, but its function, or that of any imprinted gene, has not been investigated in any marsupial. This study examines the role of IGF2 in the yolk sac placenta of the tammar wallaby, Macropus eugenii. Results: IGF2 mRNA and protein were produced in the marsupial placenta. Both IGF2 receptors were present in the placenta, and presumably mediate IGF2 mitogenic actions. IGF2 mRNA levels were highest in the vascular region of the yolk sac placenta. IGF2 increased vascular endothelial growth factor expression in placental explant cultures, suggesting that IGF2 promotes vascularisation of the yolk sac. Conclusion: This is the first demonstration of a physiological role for any imprinted gene in marsupial placentation. The conserved imprinting of IGF2 in this marsupial and in all eutherian species so far investigated, but not in monotremes, suggests that imprinting of this gene may have originated in the placenta of the therian ancestor.
MeSH term(s)	Actins/metabolism ; Animals ; Blotting, Western ; Embryo, Mammalian/metabolism ; Female ; Gene Expression Regulation, Developmental ; Genomic Imprinting ; Immunohistochemistry ; Insulin-Like Growth Factor II/genetics ; Insulin-Like Growth Factor II/metabolism ; Marsupialia/genetics ; Marsupialia/metabolism ; Placenta/metabolism ; Pregnancy ; RNA, Messenger/genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Vascular Endothelial Growth Factor A/genetics ; Vascular Endothelial Growth Factor A/metabolism ; Yolk Sac/metabolism
Chemical Substances	Actins ; RNA, Messenger ; Vascular Endothelial Growth Factor A ; Insulin-Like Growth Factor II (67763-97-7)
Language	English
Publishing date	2008-02-20
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ISSN	1471-213X
ISSN (online)	1471-213X
DOI	10.1186/1471-213X-8-17
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