Proliferation Of Metastatic Cancer Cells Research Articles

E-cadherin Induces Serine Synthesis to Support Progression and Metastasis of Breast Cancer.

The loss of E-cadherin, an epithelial cell adhesion molecule, has been implicated in metastasis by mediating the epithelial-mesenchymal transition (EMT), which promotes invasion and migration of cancer cells. However, recent studies have demonstrated that E-cadherin supports the survival and proliferation of metastatic cancer cells. Here, we identified a metabolic role for E-cadherin in breast cancer by upregulating the de novo serine synthesis pathway (SSP). The upregulated SSP provided metabolic precursors for biosynthesis and resistance to oxidative stress, enabling E-cadherin+ breast cancer cells to achieve faster tumor growth and enhanced metastases. Inhibition of PHGDH, a rate-limiting enzyme in the SSP, significantly and specifically hampered proliferation of E-cadherin+ breast cancer cells and rendered them vulnerable to oxidative stress, inhibiting their metastatic potential. These findings reveal that E-cadherin reprograms cellular metabolism, promoting tumor growth and metastasis of breast cancers.

Blue Light Inhibits Proliferation of Metastatic Cancer Cells by Regulating Translational Initiation: A Synergistic Property with Anticancer Drugs.

The aim of this study was to examine the inhibitory effect of blue light (BL) on the proliferation of metastatic cancer cells and synergistic properties with chemo-drugs. BL significantly inhibited the proliferation of B cell lymphoma (A20 and RAMOS) cells in a dose-dependent manner. Anti-proliferative effect of BL irradiation was identified to be associated with the inhibition of proliferating-cell nuclear antigen expression and cell cycle by decreasing S-phase cells. Consistent with its inhibitory effects, BL irradiation at 20 J/cm2 daily for 10 days inhibited metastasis of cancer cells which were distributed and invaded to other organs including bone marrow, liver, kidney, etc., and induced paraplegia, thereby leading to an increased survival rate of tumor-bearing mice. Anti-proliferative activity of BL was expanded in solid tumor cells including pancreatic carcinoma (Mia PaCa-2, PANC-1), lung carcinoma A549 and colorectal carcinoma HCT116 cells. Additionally, combination with chemo-drugs such as 5-FU and gemcitabine resulted in an increase in the anti-proliferative activity after BL irradiation accompanied by regulating mRNA translational process via inhibition of p70S6K, 4EBP-1 and eIF4E phosphorylation during cellular proliferation. These results indicate the anti-metastatic and photo-biogoverning abilities of BL irradiation as a potent therapeutic potential for repressing the progression of tumor cells.

HAO1-mediated oxalate metabolism promotes lung pre-metastatic niche formation by inducing neutrophil extracellular traps

Metabolic reprogramming has been shown to be involved in cancer-induced pre-metastatic niche (PMN) formation, but the underlying mechanisms have been insufficiently explored. Here, we showed that hydroxyacid oxidase 1 (HAO1), a rate-limiting enzyme of oxalate synthesis, was upregulated in the alveolar epithelial cells of mice bearing metastatic breast cancer cells at the pre-metastatic stage, leading to oxalate accumulation in lung tissue. Lung oxalate accumulation induced neutrophil extracellular trap (NET) formation by activating NADPH oxidase, which facilitated the formation of pre-metastatic niche. In addition, lung oxalate accumulation promoted the proliferation of metastatic cancer cells by activating the MAPK signaling pathway. Pharmacologic inhibition of HAO1 could effectively suppress the lung oxalate accumulation induced by primary cancer, consequently dampening lung metastasis of breast cancer. Breast cancer cells induced HAO1 expression and oxalate accumulation in alveolar epithelial cells by activating TLR3-IRF3 signaling. Collectively, these findings underscore the role of HAO1-mediated oxalate metabolism in cancer-induced lung PMN formation and metastasis. HAO1 could be an appealing therapeutic target for preventing lung metastasis of cancer.

Inhibition of Liver Metastasis in Colorectal Cancer by Targeting IL-13/IL13Rα2 Binding Site with Specific Monoclonal Antibodies.

Simple SummaryIL13Rα2 has been repeatedly reported as an excellent therapeutic target for multiple types of advanced cancers. However, previous IL13Rα2 targeting attempts have been mostly unsuccessful. Here, we describe a novel strategy based on the blocking of the IL-13 tumorigenic activity using a highly preserved D1 peptide selected from the IL13Rα2 binding site for mouse immunization and the inhibition of the cell invasion capacity for antibody screening. The IL13Rα2 D1 peptide-specific monoclonal antibody 5.5.4 has demonstrated a large capacity for blocking IL13Rα2 signaling capacity and protecting mice against established and non-established liver metastasis in colorectal cancer. These positive results predict a potential application to other IL13Rα2 positive cancers.Background: IL13Rα2 is reportedly a promising therapeutic target in different cancers. Still, no specific antagonists have reached the clinics yet. We investigated the use of a IL-13/IL13Rα2 binding motif, called D1, as a new target for the development of therapeutic monoclonal antibodies (mAbs) for colorectal cancer (CRC) metastasis. Methods: IL13Rα2 D1 peptides were prepared and used for immunization and antibody development. Antibodies were tested for inhibition of cellular invasion through Matrigel using CRC cell lines. Effects of the mAbs on cell signaling, receptor internalization and degradation were determined by western blot and flow cytometry. Swiss nude mice were used for survival analysis after treatment with IL13Rα2-specific mAbs and metastasis development. Results: IL13Rα2 D1 peptides were used to generate highly selective mAbs that blocked IL13/IL13Rα2-mediated SRC activation and cell invasion in colorectal cancer cells. Antibodies also provoked a significant reduction in cell adhesion and proliferation of metastatic cancer cells. Treatment with mAbs impaired the FAK, SRC and PI3K/AKT pathway activation. Blocking effectivity was shown to correlate with the cellular IL13Rα2 expression level. Despite mAb 5.5.4 partially blocked IL-13 mediated receptor internalization from the cancer cell surface it still promotes receptor degradation. Compared with other IL13Rα2-specific antibodies, 5.5.4 exhibited a superior efficacy to inhibit metastatic growth in vivo, providing a complete mouse survival in different conditions, including established metastasis. Conclusions: Monoclonal antibody 5.5.4 showed a highly selective blocking capacity for the interaction between IL-13 and IL13Rα2 and caused a complete inhibition of liver metastasis in IL13Rα2-positive colorectal cancer cells. This capacity might be potentially applicable to other IL13Rα2-expressing tumors.

Lipid-Functionalized Graphene Loaded with hMnSOD for Selective Inhibition of Cancer Cells.

Combination therapies utilize multiple mechanisms to target cancer cells to minimize cancer cell survival. Graphene provides an ideal platform for combination therapy due to its photothermal properties and high loading capacity for cancer-fighting molecules. Lipid functionalization of graphene extends its potential as a therapeutic platform by improving its biocompatibility and functionality. Previous studies involving graphene demonstrated its usage as a therapeutic vehicle; however, the effect of bare and engineered graphene structures on oxidative stress has not been comprehensively investigated. Because oxidative stress has been linked to cancer progression, it is vital to examine the generation of reactive oxygen species (ROS) in response to therapeutic platforms. This study functionalizes reduced graphene oxide (rGO) with lipids and the antioxidant enzyme human manganese superoxide dismutase (hMnSOD) and presents a detailed characterization of cellular responses to bare and functionalized rGO nanostructures in tumorigenic and nontumorigenic breast cell lines. Each cell type displayed distinct responses depending on whether they were normal, nonmetastatic, or metastatic cells. Bare rGO significantly reduced cell growth and substantially increased ROS production in all cell lines and instigated necrosis in metastatic breast cancer cells. Cell proliferation decreased in cancerous breast cells upon introduction of lipid-rGO, which correlated with peroxidation of lipids coating the rGO. In contrast, lipid-rGO nanostructures had minimal impact on proliferation and lipid peroxidation for normal breast cells. Lipid-rGO nanostructures with bound hMnSOD inhibited the proliferation of metastatic cancer cells while preventing necrosis and avoiding the negative side effects on normal cells associated with chemotherapeutic agents. Together, the results confirm the importance of functionalizing rGO for therapeutic applications and present an additional modality for the usage of graphene to selectively target cancer cells.

The CXCL5/CXCR2 axis is sufficient to promote breast cancer colonization during bone metastasis

Bone is one of the most common sites for metastasis across cancers. Cancer cells that travel through the vasculature and invade new tissues can remain in a non-proliferative dormant state for years before colonizing the metastatic site. Switching from dormancy to colonization is the rate-limiting step of bone metastasis. Here we develop an ex vivo co-culture method to grow cancer cells in mouse bones to assess cancer cell proliferation using healthy or cancer-primed bones. Profiling soluble factors from conditioned media identifies the chemokine CXCL5 as a candidate to induce metastatic colonization. Additional studies using CXCL5 recombinant protein suggest that CXCL5 is sufficient to promote breast cancer cell proliferation and colonization in bone, while inhibition of its receptor CXCR2 with an antagonist blocks proliferation of metastatic cancer cells. This study suggests that CXCL5 and CXCR2 inhibitors may have efficacy in treating metastatic bone tumors dependent on the CXCL5/CXCR2 axis.

Prognostic value of soluble angiotensin II receptor 1 and soluble angiotensin converting enzyme (CD 143) in patients with acute leukemia

AbstractBackgroundThe renin-angiotensin system (RAS) is a bioenzymic cascade that plays an integral role in cardiovascular homoeostasis by influencing vascular tone, fluid and electrolyte balance and the sympathetic nervous system. RAS was viewed as a circulating endocrine system, whereby renin released from the juxtaglomerular cells of the kidney cleaves the liver-derived macroglobulin precursor angio-tensinogen, to produce the inactive decapeptide angiotensin I, which is then converted to the active octapeptide Ang II by angiotensin converting enzyme (ACE) within the pulmonary. There is increasing evidence that Ang II, a major regulator of blood pressure and cardiovascular homeostasis, is involved in the regulation of cell proliferation, angiogenesis, inflammation and tissue remodeling, which suggests that this peptide might also play a role in cancer. Ang II is the main effector of the RAS and it alternatively binds to either Ang II T1R or Ang II T2R. The Ang II T1R and Ang II T2R can act as antagonists, and mediate effects on cell migration and proliferation of metastatic cancer cells and hemopoietic stem-progenitor cells. Components of the RAS are frequently differentially expressed in various cancers in comparison with their corresponding non- malignant tissue. Yet, the RAS has not been fully elucidated in patients with acute leukemia. Objective: The aim of the present work was to study serum level of Angiotensin II receptor type 1 and the soluble angiotensin converting enzyme (CD143) in patients with acute leukemia in order to extrapolate their possible prognostic value.Subjects and MethodsThe present study included 20 healthy volunteers clinically free from hypertension or sarcoidosis, 20 patients of newly diagnosed AML and 20 patients of newly diagnosed ALL. Blood samples were collected from all subjects and the level of serum ACE and serum Ang IIT1R were measured by enzyme linked immunossorbent assay.ResultsThe activity of ACE (U/L) and the concentration of Ang IIT1R (U/L) in patients groups with either AML or ALL before therapy were significantly higher than in control group. After therapy, the activity of the enzyme and its receptor concentration in both groups of patients were significantly decreased but still significantly higher than in normal control subjects.ConclusionsEstimating the serum level of ACE and soluble Ang IIT1R is of informative diagnostic and prognostic value. Estimation serum level of ACE and Ang IIT1R levels in patients with either AML or ALL is of value in deciding the treatment protocol.

Monoclonal Antibodies Directed against Cadherin RGD Exhibit Therapeutic Activity against Melanoma and Colorectal Cancer Metastasis.

Purpose: New targets are required for the control of advanced metastatic disease. We investigated the use of cadherin RGD motifs, which activate the α2β1integrin pathway, as targets for the development of therapeutic monoclonal antibodies (mAb).Experimental Design: Cadherin 17 (CDH17) fragments and peptides were prepared and used for immunization and antibody development. Antibodies were tested for inhibition of β1 integrin and cell adhesion, proliferation, and invasion assays using cell lines from different cancer types (colorectal, pancreatic, melanoma, and breast cancer). Effects of the mAbs on cell signaling were determined by Western blot analysis. Nude mice were used for survival analysis after treatment with RGD-specific mAbs and metastasis development.Results: Antibodies against full-length CDH17 failed to block the binding to α2β1 integrin. However, CDH17 RGD peptides generated highly selective RGD mAbs that blocked CDH17 and vascular-endothelial (VE)-cadherin-mediated β1 integrin activation in melanoma and breast, pancreatic, and colorectal cancer cells. Antibodies provoked a significant reduction in cell adhesion and proliferation of metastatic cancer cells. Treatment with mAbs impaired the integrin signaling pathway activation of FAK in colorectal cancer, of JNK and ERK kinases in colorectal and pancreatic cancers, and of JNK, ERK, Src, and AKT in melanoma and breast cancer. In vivo, RGD-specific mAbs increased mouse survival after inoculation of melanoma and colorectal cancer cell lines to cause lung and liver metastasis, respectively.Conclusions: Blocking the interaction between RGD cadherins and α2β1 integrin with highly selective mAbs constitutes a promising therapy against advanced metastatic disease in colon cancer, melanoma, and, potentially, other cancers. Clin Cancer Res; 24(2); 433-44. ©2017 AACRSee related commentary by Marshall, p. 253.

Exosome-delivered EGFR regulates liver microenvironment to promote gastric cancer liver metastasis

The metastatic organotropism has been one of the cancer’s greatest mysteries since the ‘seed and soil’ hypothesis. Although the role of EGFR in cancer cells is well studied, the effects of secreted EGFR transported by exosomes are less understood. Here we show that EGFR in exosomes secreted from gastric cancer cells can be delivered into the liver and is integrated on the plasma membrane of liver stromal cells. The translocated EGFR is proved to effectively activate hepatocyte growth factor (HGF) by suppressing miR-26a/b expression. Moreover, the upregulated paracrine HGF, which binds the c-MET receptor on the migrated cancer cells, provides fertile ‘soil’ for the ‘seed’, facilitating the landing and proliferation of metastatic cancer cells. Thus, we propose that EGFR-containing exosomes derived from cancer cells could favour the development of a liver-like microenvironment promoting liver-specific metastasis.

Abstract 3349: Crosstalk between Runx2 regulatory network and insulin-like growth factor receptor/Akt signaling pathway in bone metastasis of breast cancer

Abstract The metastasis of breast cancer cells to bones is a leading cause of cancer-related deaths. The bone microenvironment presents a vital source of several growth factors that could promote survival and proliferation of metastatic cancer cells. The bone-derived insulin-like growth factor-1 (IGF-1) binds to insulin-like growth factor receptor (IGF-1R) to initiate intracellular signaling. The downstream signaling events in response to IGF-1R stimulation include activation of Phosphatidyl Inositol 3’ Kinase (PI3K) and Mitogen Activated Protein Kinase (MAPK) pathways. However, the inhibition of these signaling pathways has shown modest benefit in recent clinical trials necessitating the need to identify regulatory mechanisms to successfully block breast cancer-associated bone metastasis. The Runt-related transcription factor, Runx2, is required for normal bone development but is also aberrantly expressed at higher levels in invasive mammary epithelial cells and promotes invasive properties. Previously, we have identified a mechanism wherein Runx2 promotes survival of parental population of invasive breast cancer MDA-MB-231cells by promoting phosphorylation of serine/threonine kinase Akt (pAkt) via mammalian target of rapamycin complex-2 proteins. To determine the implications of Runx2-mediated pAkt regulation in bone metastasis of breast cancers, we harvested Runx2 knockdown or control MDA-MB-231 cells from bones 7 weeks post intracardiac injections in NOD/SCID mice. The gene and protein expression levels of IGF-1R/Akt pathway effectors in the bone seeking subpopulation were determined by real-time PCR and western blotting approaches. The bone-derived Runx2 knockdown cells showed increased IGF-1Rβ levels compared to controls. The Runx2 knockdown MDA-MB-231 cells showed an increase in pIGF1R-β and pAkt levels in response to IGF-1 stimulation that could be inhibited with IGF-1Rβ inhibitor OSI-906 (Linsitinib). However, despite inhibition of IGF-1R and Akt activity with OSI-906 treatment, no alterations were observed in growth or survival of bone-seeking cells. Interestingly, in contrast to IGF-1Rβ and pAkt, the Erk 1/2 levels were reduced in bone-derived Runx2 knockdown MDA-MB-231 cells compared to parental MDA-MB-231 cells. The treatment of cells with Erk 1/2 inhibitor PD184161 reduced cell proliferation and clonogenic potential. Furthermore, the Runx2 protein levels were also reduced in bone seeking subpopulation compared to parental MDA-MB-231cells. Taken together, our studies indicate Runx2-mediated suppression of IGF-1Rβ and Akt but upregulation of Erk signaling in bone seeking breast cancer cells compared to parental MDA-MB-231 cells. Our results suggest that the growth of bone metastatic cancer cells could be effectively blocked by inhibition of Runx2 and Erk pathway. Citation Format: Manish Tandon, Zujian Chen, Jitesh Pratap. Crosstalk between Runx2 regulatory network and insulin-like growth factor receptor/Akt signaling pathway in bone metastasis of breast cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3349. doi:10.1158/1538-7445.AM2014-3349

Correction for Shibue and Weinberg, Integrin β1-focal adhesion kinase signaling directs the proliferation of metastatic cancer cells disseminated in the lungs

Correction for Shibue and Weinberg, Integrin β1-focal adhesion kinase signaling directs the proliferation of metastatic cancer cells disseminated in the lungs

New Wrinkle Between Cancer and Blood Coagulation: Metastasis and Cleavage of von Willebrand Factor by ADAM28

New Wrinkle Between Cancer and Blood Coagulation: Metastasis and Cleavage of von Willebrand Factor by ADAM28

Abstract 3251: Characterization of a novel Rac inhibitor as an anti cancer metastasis compound

Abstract The objective of this project is to design novel therapeutics targeting cancer metastasis, the most deadly aspect of epithelial cancers. Our previous studies have shown that Rac, a key signaling intermediate that regulates actin cytoskeletal changes during cell migration/invasion and cancer metastasis, is associated with high cancer metastatic efficiency. Since Rac proteins are activated by GDP/GTP exchange, inhibition of the interaction of Rac with its guanine nucleotide exchange factors (GEF) is expected to inhibit Rac activity. Since the known Rac inhibitor NSC-23766 is not effective at therapeutically useful concentrations, we synthesized novel NSC-23766 derivatives. Results show that the new inhibitor EHop-016 is 100-fold more efficient than the parent compound at inhibiting Rac activity of metastatic cancer cells without affecting the activity of the related GTPase, Rho. EHop-016 inhibited Rac activity of metastatic cancer cells with an IC50 of 0.78 microM. Cell viability assays, performed to test for the toxicity of this compound, demonstrated that at concentrations below 5 microM, EHop-016 did not affect mammary epithelial cell (MCF-10A) viability and decreased proliferation of metastatic cancer cells (MDA-MB-435) by only 20%. We conclude that EHop-16 is a potent Rac inhibitor with promise of further development as a small molecule inhibitor of cancer cell invasion, and thus, metastasis. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 3251. doi:10.1158/1538-7445.AM2011-3251

Correction for Shibue and Weinberg, Integrin β 1 -focal adhesion kinase signaling directs the proliferation of metastatic cancer cells disseminated in the lungs

Correction for Shibue and Weinberg, Integrin β ₁ -focal adhesion kinase signaling directs the proliferation of metastatic cancer cells disseminated in the lungs

Integrin β 1 -focal adhesion kinase signaling directs the proliferation of metastatic cancer cells disseminated in the lungs

The development of metastases is an extended and inefficient process involving multiple steps. The last of these involves the growth of micrometastases into macroscopic tumors. We show here that intravenously injected, nonmetastatic cancer cells cease proliferating after extravasating into the parenchyma of the lungs; this response is attributable to the cells inability to trigger adhesion-related signaling events when they are scattered sparsely within the extracellular matrix (ECM) of the parenchyma. We recapitulate this situation by culturing these nonmetastatic cells at low seeding density in ECM-derived gels in vitro, in which they undergo cell-cycle arrest resulting, in part, from insufficient activation of focal adhesion kinase (FAK). Metastatic cancer cells, in contrast, show sufficient FAK activation to enable their proliferation within ECM gels in vitro and continue cell-cycle progression within the lung parenchyma in vivo. Activation of FAK in these metastatic cells depends on expression of the beta(1) subunit of integrins, and proliferation of these cells after extravasation in the lungs is diminished by knocking down the expression of either FAK or integrin beta(1). These results demonstrate the critical role of integrin beta(1)-FAK signaling axis in controlling the initial proliferation of micrometastatic cancer cells disseminated in the lungs.

Identification of a unique set of genes altered during cell-cell contact in an in vitro model of prostate cancer bone metastasis

The interaction between prostate cancer cells and bone marrow stromal cells (BMSC) is critical for survival and proliferation of metastatic cancer cells in the bone microenvironment. In order to study molecular mechanisms of prostate cancer bone metastasis, we established a novel heterotypic co-culture system, in which the role of direct cell-cell contact between prostate cancer cells and BMSC in addition to soluble factors can be analyzed. Using both bi-compartmental (insert) system and heterotypic (contact) system, we identified gene expression profiles of interaction between prostate cancer and bone cells. Analysis of differential gene expressions in these two co-culture systems revealed three distinctive sets of genes: 1) genes that were modified only by soluble factors; 2) genes that were regulated by both soluble factors and physical contact; and 3) genes that were altered only by physical contact. The last group consisted of specific set of genes including collagen III, IV, X, XII, integrin alpha1, alpha2, MMP-2, MMP-9, uPA, biglycan, osteopontin and raf-1 in PC3, and collagen VIII, IX, BMP6, TGFbeta1, Smad6 and Twist in BMSC. Among genes that were modified by both soluble factors and physical contact, the gene expression was affected in the same direction (such as MKK4) or in the opposite direction (such as TGFbeta receptor 3). Overall, this suggests that heterotypic cell-cell contact may act as an independent factor affecting the progression of bone metastasis.