TPA‐enhanced invasion of matrigel associated with augmentation of cell motility but not metalloproteinase activity in a highly metastatic variant (L‐10) of human rectal adenocarcinoma cell line RCM‐1

Nabeshima, Kazuki; Komada, Naoto; Kishi, Jun-Ichi; Koita, Hiroyuki; Inoue, Teruhiko; Hayakawa, Taro; Koono, Masashi

doi:10.1002/ijc.2910550617

Cited by 19 publications

(12 citation statements)

References 21 publications

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“…In this regard, the metastatic potential of B16 melanoma cells has been correlated with PKC activity and can be considerably enhanced with PKC activators [55]. Similar results have been reported for Lewis lung carcinoma cells [56], mouse mammary adenocarcinoma cell line [57], and more recently, for human colorectal [58], bladder [37], and gastric [59] carcinomas. However, this report is the first to suggest that there is a direct relationship between PKC and invasion in gliomas.…”

Section: Discussionsupporting

confidence: 69%

Glioma invasionin vitro: regulation by matrix metalloprotease-2 and protein kinase C

et al. 1996

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A hallmark of invasive tumors is their ability to effect degradation of the surrounding extracellular matrix (ECM) by the local production of proteolytic enzymes, such as the matrix metalloproteases (MMPs). In this paper, we demonstrate that the invasion of human gliomas is mediated by a 72 kDa MMP, referred to as MMP-2, and provide further evidence that the activity of MMP-2 is regulated by protein kinase C (PKC). The invasiveness of five human glioma cell lines (A172, U87, U118, U251, U563) was assessed in an in vitro invasion assay and was found to correlate with the level of MMP-2 activity (r2 = 0.95); in contrast, the activity of this 72 kDa metalloprotease was barely detectable in non-invasive control glial cells (non-transformed human astrocytes and oligodendrocytes). Treatment with 1,10-phenanthroline, a metalloprotease inhibitor, or with a synthetic dipeptide, containing a blocking sequence (ala-phe) specific for MMPs, resulted in a > 90% reduction in glioma invasion. Furthermore, this MMP-2 activity could be inhibited by the treatment of tumor cells with calphostin C, a specific inhibitor of PKC. Glioma cell lines treated with calphostin C demonstrated a dose-dependent decrease (IC50 = 30 nM) in tumor invasiveness with a concomitant reduction in the activity of the MMP-2. Conversely, treatment of non-invasive control astrocytes with a PKC activator (phorbol ester) led to a corresponding increase in their invasiveness and metalloprotease activity. These findings support the postulate that MMP-2 activity constitutes an important effector of human glioma invasion and that the regulation of this proteolytic activity can be modulated by PKC.

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Section: Discussionsupporting

confidence: 69%

Glioma invasionin vitro: regulation by matrix metalloprotease-2 and protein kinase C

et al. 1996

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“…The cells showed a peak motile response at 20 ng/ml of TPA [8]. The migration was time dependent up to 24 h and approximately 80% of the maximum migration occurred by 6 h after TPA treatment [9].…”

Section: Discussionmentioning

confidence: 99%

“…The motility assay was run as described previously [8] with some modifications. Briefly, L-10 cells were seeded into compartments of an eight-well Lab-Tek tissue culture chamber slide (1.5 x 105 cells in 0.4 ml growth medium) and allowed to attach for 24 h at standard culture conditions (37°C, 5% CO2 in air, 100% humidity).…”

Section: Cell Motility On a Lab-tek Chamber Slidementioning

confidence: 99%

“…The migrating cells did not overlap one another but formed coherent cell sheets. Since TPA treatment induced this L-10 cell translocation with the peak response at 20 ng/ml [8], stimulation with 20 ng/ml TPA was performed if not otherwise indicated.…”

Section: Cell Motility On a Lab-tek Chamber Slidementioning

confidence: 99%

“…This TPA-enhanced invasion of Matrigel was associated with augmentation of cell motility but not metalloproteinase activity [8]. To elucidate the mechanism of this enhanced L-10 cell motility we used a simple Lab-Tek chamber motility assay, which meets the requirements for morphological observation [9].…”

Section: Introductionmentioning

confidence: 99%

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Ultrastructural study of TPA-induced cell motility: human well-differentiated rectal adenocarcinoma cells move as coherent sheets via localized modulation of cell-cell adhesion

et al. 1995

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We previously found that 12-O-tetradecanoylphorbol-13-acetate (TPA)-enhanced invasion of Matrigel was associated with augmentation of cell motility but not with metalloproteinase activity in a highly metastatic variant (L-10) of human rectal adenocarcinoma cell line RCM-1. In a two-dimensional cell motility assay, TPA induced active L-10 cell locomotion with characteristic morphology; the cells moved outwards from the cell islands mainly as a localized coherent sheet of cells. The leading cells showed locomotor morphologies with fan-shaped leading lamellae while the following cells had cell contacts on all sides and appeared to lack leading lamellae. In the present ultrastructural study, the following cells frequently showed tapering cytoplasmic protrusions and leading lamella-like processes underlapping a preceding cell, indicating that the locomotion mechanism is almost the same for both the leading and following cells. For this type of locomotion as a coherent sheet we propose that localized modulation of cell-cell adhesion was induced such that wide intercellular gaps occurred at the lower portion of the cells to allow the cells to extend the tapering cytoplasmic processes and leading lamellae while close cell-cell contacts remained at the upper portion of the cells. These TPA-induced changes took place predominantly in the cells at the periphery of the cell islands, while the cells in the middle of the cell islands maintained close cell-cell contacts including complex interdigitation all around the cells, suggesting the modulation of TPA action by cell-cell interaction. Additionally, consistent with the evidence for junctional complexes between the cells moving outwards, the Lucifer-yellow dye transfer studies showed some, limited cell-cell coupling, suggesting the presence of at least some gap junctional intercellular communication in the moving cell sheets.

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PMA-induced reduction in invasiveness is associated with hyperphosphorylation of MARCKS and talin in invasive bladder cancer cells

Yokoyama¹,

Ito²,

Hanson³

et al. 1998

Int. J. Cancer

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Protein kinase C (PKC) plays a critical role in signal transduction for a variety of cell activation processes. Enhanced PKC activity is often found in cancer cells that show marked invasive and/or metastatic potential. Thus, a specific PKC inhibitor may serve as a tool to reduce invasive or metastatic potential of cancer cells. We show here that phorbol 12‐myristate 13‐acetate (PMA), a PKC activator, also reduces invasiveness of EJ invasive transitional carcinoma cells. PMA‐induced reduction in invasiveness was parallel with inhibition of cell motility. PMA neither induced E‐cadherin expression nor augmented cell‐matrix adhesion of EJ cells. PMA caused retraction of microspikes from the rim of the cells and consequently rounding of the cellular rim, and the disappearance of microfilaments from the cytoplasm. PMA at 10−7 M, at which concentration the motility of EJ cells was completely inhibited, down‐regulated PKC activity over 5 hr after transient translocation of PKC activity to the membrane fraction. At the same time, PMA induced hyperphosphorylation of MARCKS and talin. During the process of cell movement, actin‐binding proteins are in a cycle of phosphorylation and dephosphorylation. Once this cycle is interrupted, cells can no longer maintain the dynamics of cytoskeletal structure. We suggest that retention of the hyperphosphorylated state of MARCKS and talin is responsible for the mechanism(s) by which PMA produces inhibitory activity against invasiveness of EJ cells. Int. J. Cancer 75:774–779, 1998.© 1998 Wiley‐Liss, Inc.

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TPA‐enhanced invasion of matrigel associated with augmentation of cell motility but not metalloproteinase activity in a highly metastatic variant (L‐10) of human rectal adenocarcinoma cell line RCM‐1

Cited by 19 publications

References 21 publications

Glioma invasionin vitro: regulation by matrix metalloprotease-2 and protein kinase C

Glioma invasionin vitro: regulation by matrix metalloprotease-2 and protein kinase C

Ultrastructural study of TPA-induced cell motility: human well-differentiated rectal adenocarcinoma cells move as coherent sheets via localized modulation of cell-cell adhesion

PMA-induced reduction in invasiveness is associated with hyperphosphorylation of MARCKS and talin in invasive bladder cancer cells

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