Blockade of Tumor Cell Transforming Growth Factor-βs Enhances Cell Cycle Progression and Sensitizes Human Breast Carcinoma Cells to Cytotoxic Chemotherapy

doi:10.1006/excr.1998.4261

Experimental Cell Research

Volume 245, Issue 2, 15 December 1998, Pages 350-359

https://doi.org/10.1006/excr.1998.4261 Get rights and content

Abstract

We have examined the effect of neutralizing TGF-β antibodies on cisplatin-mediated cytotoxicity against MDA-231 human breast tumor cell spheroids. These tridimensionalin vitrosystems have been shown to recapitulate the drug sensitivity pattern of tumor cellsin vivo.MDA-231 tumor cell spheroids exhibit higher protein levels of the cyclin-dependent kinase (Cdk) inhibitors p21 and p27 and >10-fold lower Cdk2 activity compared to adherent cell monolayers, as well as pRb hypophosphorylation, a predominant G1 population, and a cisplatin 1-h IC₅₀of approximately 100 μM. Treatment of MDA-231 cells in monolayer with cisplatin for 1 h, subsequently grown as spheroids, increased steady-state TGF-β1 mRNA levels, secretion of active TGF-β, cellular Cdk2 activity, pRb phosphorylation, and p21 protein levels, while downregulating p27. Accumulation of cells in G2M and progression into S were noted 48 h after treatment with 100 μM cisplatin. We tested whether drug-induced upregulation of TGF-β1 and p21, perhaps by preventing cell cycle progression, were protective mechanisms against drug-mediated toxicity by using neutralizing anti-TGF-β antibodies. Anti-TGF-β antibodies diminished the induction of p21, enhanced the activation of Cdk2, and facilitated progression into S and G2M following cisplatin treatment. This resulted in a >twofold enhancement of drug-induced DNA fragmentation and a shift in the cisplatin 1-h IC₅₀from 100 to <10 μM. These data suggest that tumor cell TGF-β1 may protect from DNA damage and that postchemotherapy administration of TGF-β inhibitors may facilitate progression beyond G1/S, potentially increasing the efficacy of cytotoxic chemotherapy.

References (58)

R. Baillie et al.
Multiple forms of TGF-β in breast tissue: A biologically active form of the small latent complex of TGF-β1
Eur. J. Cancer
(1996)
Y. Kakeji et al.
Dynamics of tumor oxygenation, CD31 staining and transforming growth factor-β levels after treatment with radiation or cyclophosphamide in the rat 13762 mammary carcinoma
Int. J. Radiat. Oncol. Biol. Phys.
(1997)
R.M. Hoffman
In vitro assays for chemotherapy sensitivity
Crit. Rev. Oncol. Hematol.
(1993)
J.M. Yingling et al.
Signaling by the transforming growth factor-β receptors
Biochim. Biophys. Acta
(1995)
Y. Remvikos et al.
Cell cycle modifications of breast cancer during neoadjuvant chemotherapy: A flow cytometry study of fine needle aspirates
Eur. J. Cancer
(1993)
H.L. Moses
The biological action of transforming growth factor β
Growth Factors from Genes to Clinical Application
(1990)
Roberts, A. B. Sporn, M. B. 1990, The transforming growth factor-βs, In, Handbook of Experimental Pharmacology, 95,...
M.G. Alexandrow et al.
Transforming growth factor β and cell cycle regulation
Cancer Res.
(1995)
K.M. Koli et al.
Complex role of tumor cell transforming growth factor (TGF)-βs on breast carcinoma progression
J. Mammary Gland Biol. & Neopl.
(1996)
M.G. Brattain et al.
Defects of TGF-β receptor signaling in mammary cell tumorigenesis
J. Mammary Gland Biol. Neopl.
(1996)

S.M. Gorsch et al.

Immunohistochemical staining for transforming growth factor β₁

Cancer Res.

(1992)

A.M. Thompson et al.

Transforming growth factor β₁

Br. J. Cancer

(1991)

B.A. Teicher et al.

Transforming growth factor-β in in vivo resistance

Cancer Chemother. Pharmacol.

(1996)

B.A. Teicher et al.

Reversal of in vivo drug resistance by the transforming growth factor-β inhibitor decorin

Int. J. Cancer

(1997)

B.A. Teicher et al.

Tumor resistance to alkylating agents conferred by mechanisms operative only in vivo

Science

(1990)

H. Kobayashi et al.

Acquired multicellular-mediated resistance to alkylating agents in cancer

Proc. Natl. Acad. Sci. USA

(1993)

R. Cailleau et al.

Breast tumor cell lines from pleural effusions

J. Natl. Cancer Inst.

(1974)

B.A. Arrick et al.

Differential regulation of expression of three transforming growth factor β species in human breast cancer cell lines by estradiol

Cancer Res.

(1990)

C.L. Arteaga et al.

Growth stimulation of breast cancer cells with anti-transforming growth factor β antibodies: Evidence for negative autocrine growth regulation by transforming growth factor β

Cell Growth Differ.

(1990)

C.L. Arteaga et al.

Anti-transforming growth factor (TGF)-β antibodies inhibit breast cancer cell tumorigenicity and increase mouse spleen natural killer cell activity: Implications for a possible role of tumor cell/host TGF-β interactions in human breast cancer progression

J. Clin. Invest.

(1993)

C. Lucas et al.

The autocrine production of transforming growth factor-β₁

J. Immunol.

(1990)

M. Meyerson et al.

Identification of G₁

Mol. Cell. Biol.

(1994)

T.T.Y. Wang et al.

Effects of estrogen on apoptotic pathways in human breast cancer cell line MCF-7

Cancer Res.

(1995)

K.M. Koli et al.

Predominant cytosolic localization of type II transforming growth factor β receptors in human breast carcinoma cells

Cancer Res.

(1997)

R. Derynck et al.

Human transforming growth factor-beta cDNA sequence and expression in tumor cell lines

Nature

(1985)

F. Fang et al.

Dependence of cyclin E-CDK2 kinase activity on cell anchorage

Science

(1996)

B. St. Croix et al.

Impact of cyclin-dependent kinase inhibitor p27^Kip1

Nature Med.

(1996)

R.S. Kerbel et al.

Multicellular resistance: A new paradigm to explain aspects of acquired drug resistance of solid tumors

Cold Spring Harbor Symp. Quant. Biol.

(1994)

Cited by (72)

The transcriptional regulators TAZ and YAP direct transforming growth factor β-induced tumorigenic phenotypes in breast cancer cells
2014, Journal of Biological Chemistry
Uncontrolled transforming growth factor-β (TGFβ) signaling promotes aggressive metastatic properties in late-stage breast cancers. However, how TGFβ-mediated cues are directed to induce tumorigenic events is poorly understood, particularly given that TGFβ has clear tumor suppressing activity in other contexts. Here, we demonstrate that the transcriptional regulators TAZ and YAP (TAZ/YAP), key effectors of the Hippo pathway, are necessary to promote and maintain TGFβ-induced tumorigenic phenotypes in breast cancer cells. Interactions between TAZ/YAP, TGFβ-activated SMAD2/3, and TEAD transcription factors reveal convergent roles for these factors in the nucleus. Genome-wide expression analyses indicate that TAZ/YAP, TEADs, and TGFβ-induced signals coordinate a specific pro-tumorigenic transcriptional program. Importantly, genes cooperatively regulated by TAZ/YAP, TEAD, and TGFβ, such as the novel targets NEGR1 and UCA1, are necessary for maintaining tumorigenic activity in metastatic breast cancer cells. Nuclear TAZ/YAP also cooperate with TGFβ signaling to promote phenotypic and transcriptional changes in nontumorigenic cells to overcome TGFβ-repressive effects. Our work thus identifies cross-talk between nuclear TAZ/YAP and TGFβ signaling in breast cancer cells, revealing novel insight into late-stage disease-driving mechanisms.
Defects in recombination activity caused by somatic and germline mutations in the multimerization/BRCA2 binding region of human RAD51 protein
2017, DNA Repair
Citation Excerpt :
On the other hand ∼30% of sporadic breast tumors and cell lines exhibit down-regulation of RAD51 expression [14–17]. RAD51 down-regulation increases the radiosensitivity of prostate cancer and malignant glioma cells [18,19]. Recent studies have identified several RAD51 missense variants that are associated with cancer.
The human RAD51 recombinase possesses DNA pairing and strand exchange activities that are essential for the error-free, homology-directed repair of DNA double-strand breaks. The recombination activities of RAD51 are activated upon its assembly into presynaptic filaments on single-stranded DNA at resected DSB ends. Defects in filament assembly caused by mutations in RAD51 or its regulators such as BRCA2 are associated with human cancer. Here we describe two novel RAD51 missense variants located in the multimerization/BRCA2 binding region of RAD51. F86L is a breast tumor-derived somatic variant that affects the interface between adjacent RAD51 protomers in the presynaptic filament. E258A is a germline variant that maps to the interface region between the N-terminal and RecA homology domains of RAD51. Both variants exhibit abnormal biochemistry including altered DNA strand exchange activity. Both variants inhibit the DNA strand exchange activity of wild-type RAD51, suggesting a mechanism for negative dominance. The inhibitory effect of F86L on wild-type RAD51 is surprising since F86L alone exhibits robust DNA strand exchange activity. Our findings indicate that even DNA strand exchange-proficient variants can have negative functional interactions with wild-type RAD51. Thus heterozygous F86L or E258 mutations in RAD51 could promote genomic instability, and thereby contribute to tumor progression.
5.17 Three-dimensional bioengineered cancer models
2017, Comprehensive Biomaterials II
Cancer is an evolving, dynamic, evasive and heterogeneous disease, characterized by high morbidity and mortality. It is widely acknowledged that the tumor microenvironment plays a fundamental role in cancer development and progression. Despite this, most in vitro cancer models are simple, two-dimensional monocultures of immortalized tumor cell lines. There is an immediate need to develop three-dimensional, multicellular biomimetic in vitro constructs that can model specific stages of tumorigenesis to aid cancer research and drug discovery. This chapter aims to review the latest advances in three-dimensional bioengineered cancer modeling, with particular focus on models that retain the biological and physical characteristics of the in vivo tumor microenvironment.
RAD51 variant proteins from human lung and kidney tumors exhibit DNA strand exchange defects
2016, DNA Repair
Citation Excerpt :
RAD51 overexpression has been reported in malignant prostate cancer, small cell lung carcinoma, and invasive ductal breast cancer, where in the latter case the level of RAD51 expression correlates directly with the histological grading of the tumor [10]. Conversely, down-regulation of RAD51 is observed in approximately 30% of sporadic breast tumors and cell lines [13–16], and is reported to increase the radiosensitivity of prostate cancer and malignant glioma cells [17,18]. Sequence variants of RAD51 protein have received less study due to their low penetrance.
In human cells, error-free repair of DNA double-strand breaks requires the DNA pairing and strand exchange activities of RAD51 recombinase. Activation of RAD51 recombination activities requires the assembly of RAD51 presynaptic filaments on the single-stranded DNA that forms at resected DSB ends. Mutations in proteins that control presynaptic filament assembly, such as BRCA2, and in RAD51 itself, are associated with human breast cancer. Here we describe the properties of two mutations in RAD51 protein that derive from human lung and kidney tumors, respectively. Sequence variants Q268P and Q272L both map to the DNA binding loop 2 (L2) region of RAD51, a motif that is involved in DNA binding and in the allosteric activation of ATP hydrolysis and DNA strand exchange activities. Both mutations alter the thermal stability, DNA binding, and ATPase properties of RAD51, however both variants retain intrinsic DNA strand exchange activity towards oligonucleotide substrates under optimized conditions. In contrast, both Q268P and Q272L variants exhibit drastically reduced DNA strand exchange activity in reaction mixtures containing long homologous ssDNA and dsDNA substrates and human RPA protein. Mixtures of wild-type and variant proteins also exhibit reduced DNA strand exchange activity, suggesting that heterozygous mutations could negatively affect DNA recombination and repair processes in vivo. Together, the findings of this study suggest that hypomorphic missense mutations in RAD51 protein could be drivers of genomic instability in cancer cells, and thereby contribute to the etiology of metastatic disease.
Modeling the tumor extracellular matrix: Tissue engineering tools repurposed towards new frontiers in cancer biology
2014, Journal of Biomechanics
Citation Excerpt :
For example, the expression of important angiogenesis factors and inflammatory chemokines in 3D culture of a melanoma model were dramatically different than in 2D culture and more comparable to levels observed in in vivo human tumors (Ghosh et al., 2005). Further, cancer models in 3D show key differences in susceptibility to multiple modalities of cancer therapy including radiation, traditional chemotherapy and immune therapies (Feder-Mengus et al., 2008; Ohmori et al., 1998). 3D scaffold materials pioneered by tissue engineers not only facilitate modeling of physical and biochemical matrix cues with appropriate spatiality, but through controlled co-culture they may also help model the mixed cell populations found in different tumor compartments (epithelial vs. stromal) and different intra-tumor cell phenotypes (proliferating, apoptotic, and necrotic) that may be essential to both metastasis and methods of therapeutic resistance (Kim, 2005).
Cancer progression is mediated by complex epigenetic, protein and structural influences. Critical among them are the biochemical, mechanical and architectural properties of the extracellular matrix (ECM). In recognition of the ECM's important role, cancer biologists have repurposed matrix mimetic culture systems first widely used by tissue engineers as new tools for in vitro study of tumor models. In this review we discuss the pathological changes in tumor ECM, the limitations of 2D culture on both traditional and polyacrylamide hydrogel surfaces in modeling these characteristics and advances in both naturally derived and synthetic scaffolds to facilitate more complex and controllable 3D cancer cell culture. Studies using naturally derived matrix materials like Matrigel and collagen have produced significant findings related to tumor morphogenesis and matrix invasion in a 3D environment and the mechanotransductive signaling that mediates key tumor–matrix interaction. However, lack of precise experimental control over important matrix factors in these matrices have increasingly led investigators to synthetic and semi-synthetic scaffolds that offer the engineering of specific ECM cues and the potential for more advanced experimental manipulations. Synthetic scaffolds composed of poly(ethylene glycol) (PEG), for example, facilitate highly biocompatible 3D culture, modular bioactive features like cell-mediated matrix degradation and complete independent control over matrix bioactivity and mechanics. Future work in PEG or similar reductionist synthetic matrix systems should enable the study of increasingly complex and dynamic tumor–ECM relationships in the hopes that accurate modeling of these relationships may reveal new cancer therapeutics targeting tumor progression and metastasis.
Cisplatin-induced activation of TGF-β signaling contributes to drug resistance
2024, Oncology Research

View all citing articles on Scopus

^☆: V. SaraK. HallH. Low

¹: To whom correspondence and reprint requests should be addressed at Div. Med. Oncology/Vanderbilt University, 22nd Ave. South, 1956 TVC, Nashville, TN 37232-5536. Fax: 615-343-7602. E-mail:[email protected].

View full text

Regular ArticleBlockade of Tumor Cell Transforming Growth Factor-βs Enhances Cell Cycle Progression and Sensitizes Human Breast Carcinoma Cells to Cytotoxic Chemotherapy☆

Abstract

Eur. J. Cancer

Int. J. Radiat. Oncol. Biol. Phys.

Crit. Rev. Oncol. Hematol.

Biochim. Biophys. Acta

Eur. J. Cancer

The biological action of transforming growth factor β

Growth Factors from Genes to Clinical Application

Transforming growth factor β and cell cycle regulation

Cancer Res.

Complex role of tumor cell transforming growth factor (TGF)-βs on breast carcinoma progression

J. Mammary Gland Biol. & Neopl.

Defects of TGF-β receptor signaling in mammary cell tumorigenesis

J. Mammary Gland Biol. Neopl.

Immunohistochemical staining for transforming growth factor β1

Cancer Res.

Transforming growth factor β1

Br. J. Cancer

Transforming growth factor-β in in vivo resistance

Cancer Chemother. Pharmacol.

Reversal of in vivo drug resistance by the transforming growth factor-β inhibitor decorin

Int. J. Cancer

Tumor resistance to alkylating agents conferred by mechanisms operative only in vivo

Science

Acquired multicellular-mediated resistance to alkylating agents in cancer

Proc. Natl. Acad. Sci. USA

Breast tumor cell lines from pleural effusions

J. Natl. Cancer Inst.

Differential regulation of expression of three transforming growth factor β species in human breast cancer cell lines by estradiol

Cancer Res.

Growth stimulation of breast cancer cells with anti-transforming growth factor β antibodies: Evidence for negative autocrine growth regulation by transforming growth factor β

Cell Growth Differ.

Anti-transforming growth factor (TGF)-β antibodies inhibit breast cancer cell tumorigenicity and increase mouse spleen natural killer cell activity: Implications for a possible role of tumor cell/host TGF-β interactions in human breast cancer progression

J. Clin. Invest.

The autocrine production of transforming growth factor-β1

J. Immunol.

Identification of G1

Mol. Cell. Biol.

Effects of estrogen on apoptotic pathways in human breast cancer cell line MCF-7

Cancer Res.

Predominant cytosolic localization of type II transforming growth factor β receptors in human breast carcinoma cells

Cancer Res.

Human transforming growth factor-beta cDNA sequence and expression in tumor cell lines

Nature

Dependence of cyclin E-CDK2 kinase activity on cell anchorage

Science

Impact of cyclin-dependent kinase inhibitor p27Kip1

Nature Med.

Multicellular resistance: A new paradigm to explain aspects of acquired drug resistance of solid tumors

Cold Spring Harbor Symp. Quant. Biol.

Regular Article
Blockade of Tumor Cell Transforming Growth Factor-βs Enhances Cell Cycle Progression and Sensitizes Human Breast Carcinoma Cells to Cytotoxic Chemotherapy☆

Immunohistochemical staining for transforming growth factor β₁

Transforming growth factor β₁

The autocrine production of transforming growth factor-β₁

Identification of G₁

Impact of cyclin-dependent kinase inhibitor p27^Kip1