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A Novel Role of E-Cadherin-Based Adherens Junctions in Neoplastic Cell Dissemination.

Rubtsova SN, Zhitnyak IY, Gloushankova NA - PLoS ONE (2015)

Bottom Line: Studying interactions of IAR-6-1 transformed cells stably expressing GFP-E-cadherin with the IAR-2 epithelial monolayer, we found that IAR-6-1 cells established E-cadherin-based adhesions with normal epithelial cells: dot-like dynamic E-cadherin-based adhesions in protrusions and large adherens junctions at the cell sides and rear.IAR-6-1DNE cells expressing a dominant-negative mutant form of E-cadherin with the mutation in the first extracellular domain practically lost the ability to adhere to IAR-2 cells and invade the IAR-2 epithelial monolayer.The ability of cancer cells to form E-cadherin-based AJs with the surrounding normal epithelial cells may play an important role in driving cancer cell dissemination in the body.

View Article: PubMed Central - PubMed

Affiliation: Institute of Carcinogenesis, N.N. Blokhin Russian Cancer Research Center, Moscow, Russia.

ABSTRACT
Using confocal microscopy, we analyzed the behavior of IAR-6-1, IAR1170, and IAR1162 transformed epithelial cells seeded onto the confluent monolayer of normal IAR-2 epithelial cells. Live-cell imaging of neoplastic cells stably expressing EGFP and of normal epithelial cells stably expressing mKate2 showed that transformed cells retaining expression of E-cadherin were able to migrate over the IAR-2 epithelial monolayer and invade the monolayer. Transformed IAR cells invaded the IAR-2 monolayer at the boundaries between normal cells. Studying interactions of IAR-6-1 transformed cells stably expressing GFP-E-cadherin with the IAR-2 epithelial monolayer, we found that IAR-6-1 cells established E-cadherin-based adhesions with normal epithelial cells: dot-like dynamic E-cadherin-based adhesions in protrusions and large adherens junctions at the cell sides and rear. A comparative study of a panel of transformed IAR cells that differ by their ability to form E-cadherin-based AJs, either through loss of E-cadherin expression or through expression of a dominant negative E-cadherin mutant, demonstrated that E-cadherin-based AJs are key mediators of the interactions between neoplastic and normal epithelial cells. IAR-6-1DNE cells expressing a dominant-negative mutant form of E-cadherin with the mutation in the first extracellular domain practically lost the ability to adhere to IAR-2 cells and invade the IAR-2 epithelial monolayer. The ability of cancer cells to form E-cadherin-based AJs with the surrounding normal epithelial cells may play an important role in driving cancer cell dissemination in the body.

No MeSH data available.


Related in: MedlinePlus

Transformed IAR-6-1 epithelial cells migrate over the monolayer of normal IAR-2 epithelial cells.EGFP-expressing IAR-6-1 cells were seeded onto the confluent monolayer of mKate2-expressing IAR-2 cells. (A) A scheme of experimental design used in the present study: a glass bottom culture dish with a confluent IAR-2 monolayer (red) and transformed IAR cells (green) seeded sparsely onto the monolayer. (B) Selected frames from S2 Video with combined DIC and green channels. Asterisks indicate migration of an elongated fibroblast-like cell. Scale bar 40 μm. (C) Selected frames from S3 Video with combined red and green channels of the top confocal slices out of time lapse Z-stacks. A corresponding 450-min track (1 point/15 min) of the migrating IAR-6-1 cell is shown on Frame 6. Scale bar 20 μm.
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pone.0133578.g002: Transformed IAR-6-1 epithelial cells migrate over the monolayer of normal IAR-2 epithelial cells.EGFP-expressing IAR-6-1 cells were seeded onto the confluent monolayer of mKate2-expressing IAR-2 cells. (A) A scheme of experimental design used in the present study: a glass bottom culture dish with a confluent IAR-2 monolayer (red) and transformed IAR cells (green) seeded sparsely onto the monolayer. (B) Selected frames from S2 Video with combined DIC and green channels. Asterisks indicate migration of an elongated fibroblast-like cell. Scale bar 40 μm. (C) Selected frames from S3 Video with combined red and green channels of the top confocal slices out of time lapse Z-stacks. A corresponding 450-min track (1 point/15 min) of the migrating IAR-6-1 cell is shown on Frame 6. Scale bar 20 μm.

Mentions: Using time-lapse videomicroscopy, we investigated the behavior of transformed epithelial cells seeded onto the confluent monolayer of IAR-2 normal epithelial cells (Fig 2A). To distinguish between neoplastic and normal cells, we established stable lines of IAR-6-1, IAR1162, and IAR1170 cells expressing EGFP and IAR-2 cells expressing mKate2.


A Novel Role of E-Cadherin-Based Adherens Junctions in Neoplastic Cell Dissemination.

Rubtsova SN, Zhitnyak IY, Gloushankova NA - PLoS ONE (2015)

Transformed IAR-6-1 epithelial cells migrate over the monolayer of normal IAR-2 epithelial cells.EGFP-expressing IAR-6-1 cells were seeded onto the confluent monolayer of mKate2-expressing IAR-2 cells. (A) A scheme of experimental design used in the present study: a glass bottom culture dish with a confluent IAR-2 monolayer (red) and transformed IAR cells (green) seeded sparsely onto the monolayer. (B) Selected frames from S2 Video with combined DIC and green channels. Asterisks indicate migration of an elongated fibroblast-like cell. Scale bar 40 μm. (C) Selected frames from S3 Video with combined red and green channels of the top confocal slices out of time lapse Z-stacks. A corresponding 450-min track (1 point/15 min) of the migrating IAR-6-1 cell is shown on Frame 6. Scale bar 20 μm.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4514802&req=5

pone.0133578.g002: Transformed IAR-6-1 epithelial cells migrate over the monolayer of normal IAR-2 epithelial cells.EGFP-expressing IAR-6-1 cells were seeded onto the confluent monolayer of mKate2-expressing IAR-2 cells. (A) A scheme of experimental design used in the present study: a glass bottom culture dish with a confluent IAR-2 monolayer (red) and transformed IAR cells (green) seeded sparsely onto the monolayer. (B) Selected frames from S2 Video with combined DIC and green channels. Asterisks indicate migration of an elongated fibroblast-like cell. Scale bar 40 μm. (C) Selected frames from S3 Video with combined red and green channels of the top confocal slices out of time lapse Z-stacks. A corresponding 450-min track (1 point/15 min) of the migrating IAR-6-1 cell is shown on Frame 6. Scale bar 20 μm.
Mentions: Using time-lapse videomicroscopy, we investigated the behavior of transformed epithelial cells seeded onto the confluent monolayer of IAR-2 normal epithelial cells (Fig 2A). To distinguish between neoplastic and normal cells, we established stable lines of IAR-6-1, IAR1162, and IAR1170 cells expressing EGFP and IAR-2 cells expressing mKate2.

Bottom Line: Studying interactions of IAR-6-1 transformed cells stably expressing GFP-E-cadherin with the IAR-2 epithelial monolayer, we found that IAR-6-1 cells established E-cadherin-based adhesions with normal epithelial cells: dot-like dynamic E-cadherin-based adhesions in protrusions and large adherens junctions at the cell sides and rear.IAR-6-1DNE cells expressing a dominant-negative mutant form of E-cadherin with the mutation in the first extracellular domain practically lost the ability to adhere to IAR-2 cells and invade the IAR-2 epithelial monolayer.The ability of cancer cells to form E-cadherin-based AJs with the surrounding normal epithelial cells may play an important role in driving cancer cell dissemination in the body.

View Article: PubMed Central - PubMed

Affiliation: Institute of Carcinogenesis, N.N. Blokhin Russian Cancer Research Center, Moscow, Russia.

ABSTRACT
Using confocal microscopy, we analyzed the behavior of IAR-6-1, IAR1170, and IAR1162 transformed epithelial cells seeded onto the confluent monolayer of normal IAR-2 epithelial cells. Live-cell imaging of neoplastic cells stably expressing EGFP and of normal epithelial cells stably expressing mKate2 showed that transformed cells retaining expression of E-cadherin were able to migrate over the IAR-2 epithelial monolayer and invade the monolayer. Transformed IAR cells invaded the IAR-2 monolayer at the boundaries between normal cells. Studying interactions of IAR-6-1 transformed cells stably expressing GFP-E-cadherin with the IAR-2 epithelial monolayer, we found that IAR-6-1 cells established E-cadherin-based adhesions with normal epithelial cells: dot-like dynamic E-cadherin-based adhesions in protrusions and large adherens junctions at the cell sides and rear. A comparative study of a panel of transformed IAR cells that differ by their ability to form E-cadherin-based AJs, either through loss of E-cadherin expression or through expression of a dominant negative E-cadherin mutant, demonstrated that E-cadherin-based AJs are key mediators of the interactions between neoplastic and normal epithelial cells. IAR-6-1DNE cells expressing a dominant-negative mutant form of E-cadherin with the mutation in the first extracellular domain practically lost the ability to adhere to IAR-2 cells and invade the IAR-2 epithelial monolayer. The ability of cancer cells to form E-cadherin-based AJs with the surrounding normal epithelial cells may play an important role in driving cancer cell dissemination in the body.

No MeSH data available.


Related in: MedlinePlus