MDA-MB-435 and M14 Cell Lines: Identical but not M14 Melanoma?

Chambers, Ann F.

doi:10.1158/0008-5472.can-09-1528

Cited by 205 publications

(190 citation statements)

References 19 publications

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“…Thus, the cellular wild-type MT1-MMP, but not the inert E240A mutant and the L50D prodomain mutant, underwent the extensive intracellular self-proteolysis in MCF7 cells. Our experiments using the RFP-and GFP-tagged fluorescent biosensors also clearly demonstrated the prodomain cleavage in mammary carcinoma MCF7 and MDA-MB-435/melanoma M14 cells (23) and fibrosarcoma HT1080 cancer cells.…”

Section: Discussionsupporting

confidence: 59%

Intradomain Cleavage of Inhibitory Prodomain Is Essential to Protumorigenic Function of Membrane Type-1 Matrix Metalloproteinase (MT1-MMP) in Vivo

Golubkov

Chernov

Strongin

2011

Journal of Biological Chemistry

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Invasive cancers use pericellular proteolysis to breach the extracellular matrix and basement membrane barriers and invade the surrounding tissue. Proinvasive membrane type-1 matrix metalloproteinase (MT1-MMP) is the primary mediator of proteolytic events on the cancer cell surface. MT1-MMP is synthesized as a zymogen. The latency of MT1-MMP is maintained by its N-terminal inhibitory prodomain. In the course of MT1-MMP activation, the R 108 RKR 111 2Y 112 prodomain sequence is processed by furin. The intact prodomain released by furin alone, however, is a potent inhibitor of the emerging MT1-MMP enzyme. Evidence suggests that the prodomain undergoes intradomain cleavage at the PGD2L 50 site followed by the release of the degraded prodomain by furin cleavage that finalizes the two-step activation event. Normal and malignant cells rely on pericellular proteolysis to penetrate through tissue barriers (1-3). The invasive phenotype is frequently associated with the cellular matrix metalloproteinase (MMP) 2 activity (4). The human MMP family comprises 24 individual enzymes (5). Six membrane type MMPs (MT-MMPs) are distinguished from soluble MMPs by an additional transmembrane domain and a short cytoplasmic tail (MT1-, MT2-, MT3-, and MT5-MMP) or by a glycosylphosphatidylinositol anchor (MT4-and MT6-MMPs) (6, 7). There is compelling evidence that MT1-MMP functions as a proinvasive factor (8 -10). MT1-MMP is the primary mediator of proteolytic events on the cell surface (11). MT1-MMP is directly involved in the pericellular proteolysis of the ECM proteins, activation of soluble MMPs, and cleavage of adhesion and signaling cell receptors (12-14). Although MT1-MMP is ubiquitously expressed, its activity is elevated in metastatic malignancies (15), a cause of mortality and morbidity in cancer patients.MT1-MMP is synthesized de novo as a latent proenzyme (16). Proteolytic removal of the N-terminal prodomain is required for the proenzyme conversion into the functionally active MT1-MMP enzyme. It is established that in the course of the secretion pathway of the MT1-MMP proenzyme through the cell compartment the R 108 RKR 111 2Y 112 prodomain sequence is processed by furin (17,18). We demonstrated, however, that the intact prodomain released by furin alone is a potent inhibitor of the emerging MT1-MMP enzyme. Evidence suggests that the activation of cellular MT1-MMP represents a stepwise process (19,20). In this stepwise process, the PGD2L 50 sequence of the prodomain "bait region" is cleaved intracellularly either by MMPs, which are distinct from MT1-MMP, or by autocatalysis if cellular MT1-MMP is overexpressed. Intradomain cleavage results in the activation intermediate commencing from the N-terminal Leu 50 . Furin proteolysis then follows to complete both the degradation and removal of the residual prodomain sequence, thus transforming the activation intermediate into the mature, proteolytically active MT1-MMP enzyme commencing from the N-terminal Tyr 112 (13,21,22). We used mutagenesis to inactivate the PGD2L 50 cleavage s...

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Section: Discussionsupporting

confidence: 59%

Intradomain Cleavage of Inhibitory Prodomain Is Essential to Protumorigenic Function of Membrane Type-1 Matrix Metalloproteinase (MT1-MMP) in Vivo

Golubkov

Chernov

Strongin

2011

Journal of Biological Chemistry

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“…However, its origin became ambiguous as its gene expression profile was found to be similar to M14 cells, which were believed to have a melanoma origin. Recently, Dr Chambers reviewed all the evidence and suggested that the MDA-MB-435 cell line is still a poorly differentiated, aggressive breast cancer line [46]. Regardless of its origin, our results demonstrate that the TWIST complex is required for the MDA-MB-435 human cancer cells to invade ECM in vitro and metastasize in vivo, similar to its role in the 4T1 mouse breast cancer cells.…”

Section: The Twist/mi2/nurd/mta2 Complex Regulates Breast Cancer Metasupporting

confidence: 55%

The TWIST/Mi2/NuRD protein complex and its essential role in cancer metastasis

et al. 2010

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The epithelial-mesenchymal transition (EMT) converts epithelial tumor cells into invasive and metastatic cancer cells, leading to mortality in cancer patients. Although TWIST is a master regulator of EMT and metastasis for breast and other cancers, the mechanisms responsible for TWIST-mediated gene transcription remain unknown. In this study, purification and characterization of the TWIST protein complex revealed that TWIST interacts with several components of the Mi2/nucleosome remodeling and deacetylase (Mi2/NuRD) complex, MTA2, RbAp46, Mi2 and HDAC2, and recruits them to the proximal regions of the E-cadherin promoter for transcriptional repression. Depletion of these TWIST complex components from cancer cell lines that depend on TWIST for metastasis efficiently suppresses cell migration and invasion in culture and lung metastasis in mice. These findings not only provide novel mechanistic and functional links between TWIST and the Mi2/NuRD complex but also establish new essential roles for the components of Mi2/NuRD complex in cancer metastasis.

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“…Anti-Her2-auristatin conjugates were tested on Her2-expressing breast cancer cells for cytotoxicity (MDA-MB-435 cells transduced with Her2, and SK-BR-3 cells), and Her2 − cells (MDA-MB-435) for selectivity. MDA-MB-435/Her2 + cells were generated by stable transduction of triple negative MDA-MB-435 cells with Her2 using a lentiviral vector that contains the full Her2 transmembrane receptor gene (26), providing an isogenic comparison cell line to the original Her2 − MDA-MB-435 cell line (34)(35)(36). Anti-Her2-IgG with two nAFs conjugated at HC-A121X kills MDA-MB-435/Her2 + cells with a EC 50 of 0.37 ± 0.38 nM (mean ± SD), but does not kill MDA-MB-435/Her2 − cells (EC 50 >200 nM) ( Fig.…”

Section: Resultsmentioning

confidence: 99%

Synthesis of site-specific antibody-drug conjugates using unnatural amino acids

Axup

Bajjuri²,

Ritland³

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

509

454

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Antibody-drug conjugates (ADCs) allow selective targeting of cytotoxic drugs to cancer cells presenting tumor-associated surface markers, thereby minimizing systemic toxicity. Traditionally, the drug is conjugated nonselectively to cysteine or lysine residues in the antibody. However, these strategies often lead to heterogeneous products, which make optimization of the biological, physical, and pharmacological properties of an ADC challenging. Here we demonstrate the use of genetically encoded unnatural amino acids with orthogonal chemical reactivity to synthesize homogeneous ADCs with precise control of conjugation site and stoichiometry. p -Acetylphenylalanine was site-specifically incorporated into an anti-Her2 antibody Fab fragment and full-length IgG in Escherichia coli and mammalian cells, respectively. The mutant protein was selectively and efficiently conjugated to an auristatin derivative through a stable oxime linkage. The resulting conjugates demonstrated excellent pharmacokinetics, potent in vitro cytotoxic activity against Her2 + cancer cells, and complete tumor regression in rodent xenograft treatment models. The synthesis and characterization of homogeneous ADCs with medicinal chemistry-like control over macromolecular structure should facilitate the optimization of ADCs for a host of therapeutic uses.

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MDA-MB-435 and M14 Cell Lines: Identical but not M14 Melanoma?

Abstract: A controversy has arisen over the past several years about the true origin of the human MDA-MB-435 cell line.

Cited by 205 publications

References 19 publications

Intradomain Cleavage of Inhibitory Prodomain Is Essential to Protumorigenic Function of Membrane Type-1 Matrix Metalloproteinase (MT1-MMP) in Vivo

Intradomain Cleavage of Inhibitory Prodomain Is Essential to Protumorigenic Function of Membrane Type-1 Matrix Metalloproteinase (MT1-MMP) in Vivo

The TWIST/Mi2/NuRD protein complex and its essential role in cancer metastasis

Synthesis of site-specific antibody-drug conjugates using unnatural amino acids

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