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Fusion of CCL21 non-migratory active breast epithelial and breast cancer cells give rise to CCL21 migratory active tumor hybrid cell lines.

Berndt B, Haverkampf S, Reith G, Keil S, Niggemann B, Zänker KS, Dittmar T - PLoS ONE (2013)

Bottom Line: Cell migration demonstrated that only M13HS hybrid cell lines, but not parental derivatives, responded to CCL21 stimulation with an increased migratory activity.Knockdown of CCR7 expression by siRNA completely abrogated the CCL21 induced migration of hybrid cell lines indicating the necessity of CCL21/CCR7 signaling.Because the CCL21/CCR7 axis has been linked to metastatic spreading of breast cancer to lymph nodes we conclude from our data that cell fusion could be a mechanism explaining the origin of metastatic cancer (hybrid) cells.

View Article: PubMed Central - PubMed

Affiliation: Institute of Immunology, Center for Biomedical Education and Research (ZBAF), Witten/Herdecke University, Witten, Germany.

ABSTRACT
The biological phenomenon of cell fusion has been linked to tumor progression because several data provided evidence that fusion of tumor cells and normal cells gave rise to hybrid cell lines exhibiting novel properties, such as increased metastatogenic capacity and an enhanced drug resistance. Here we investigated M13HS hybrid cell lines, derived from spontaneous fusion events between M13SV1-EGFP-Neo breast epithelial cells exhibiting stem cell characteristics and HS578T-Hyg breast cancer cells, concerning CCL21/CCR7 signaling. Western Blot analysis showed that all cell lines varied in their CCR7 expression levels as well as differed in the induction and kinetics of CCR7 specific signal transduction cascades. Flow cytometry-based calcium measurements revealed that a CCL21 induced calcium influx was solely detected in M13HS hybrid cell lines. Cell migration demonstrated that only M13HS hybrid cell lines, but not parental derivatives, responded to CCL21 stimulation with an increased migratory activity. Knockdown of CCR7 expression by siRNA completely abrogated the CCL21 induced migration of hybrid cell lines indicating the necessity of CCL21/CCR7 signaling. Because the CCL21/CCR7 axis has been linked to metastatic spreading of breast cancer to lymph nodes we conclude from our data that cell fusion could be a mechanism explaining the origin of metastatic cancer (hybrid) cells.

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Knockdown of CCR7 expression abolish the CCL21 induced migration of M13HS hybrid cell lines.The migratory activity was analyzed using the 3D collagen matrix migration assay combined with time-lapse video-microscopy. For a better comparison the locomotory activities of CCL21 stimulated cells were calculated in relation to untreated control cells, which were set to 100%. Cells were stimulated with 500 ng/ml CCL21. Shown are the mean±SD of n = 3 independent experiments. (A) HS578T-Hyg breast cancer cells, (B) M13SV1-EGFP-Neo breast epithelial cells exhibiting stem cell characteristics, (C) M13HS-2 hybrid cells, (D) M13HS-8 hybrid cells. Statistical significance was calculated using Student's t-test: ***  =  p<0.001.
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pone-0063711-g004: Knockdown of CCR7 expression abolish the CCL21 induced migration of M13HS hybrid cell lines.The migratory activity was analyzed using the 3D collagen matrix migration assay combined with time-lapse video-microscopy. For a better comparison the locomotory activities of CCL21 stimulated cells were calculated in relation to untreated control cells, which were set to 100%. Cells were stimulated with 500 ng/ml CCL21. Shown are the mean±SD of n = 3 independent experiments. (A) HS578T-Hyg breast cancer cells, (B) M13SV1-EGFP-Neo breast epithelial cells exhibiting stem cell characteristics, (C) M13HS-2 hybrid cells, (D) M13HS-8 hybrid cells. Statistical significance was calculated using Student's t-test: ***  =  p<0.001.

Mentions: Because the CCL21/CCR7 axis has been associated with lymph node metastasis of breast cancer (for review see: [34]) cell migration studies were conducted by using the 3D-collagen matrix migration assay. In addition to naïve cells cell migration studies were also performed with cells either transfected with specific CCR7 siRNA or scrambled control RNA (scRNA). CCR7 expression data are summarized in Figure 4A and show a reduced CCR7 expression in CCR7 siRNA transfected cells, whereas CCR7 levels of scRNA transfected cells were comparable to naïve cells (Figure 4A).


Fusion of CCL21 non-migratory active breast epithelial and breast cancer cells give rise to CCL21 migratory active tumor hybrid cell lines.

Berndt B, Haverkampf S, Reith G, Keil S, Niggemann B, Zänker KS, Dittmar T - PLoS ONE (2013)

Knockdown of CCR7 expression abolish the CCL21 induced migration of M13HS hybrid cell lines.The migratory activity was analyzed using the 3D collagen matrix migration assay combined with time-lapse video-microscopy. For a better comparison the locomotory activities of CCL21 stimulated cells were calculated in relation to untreated control cells, which were set to 100%. Cells were stimulated with 500 ng/ml CCL21. Shown are the mean±SD of n = 3 independent experiments. (A) HS578T-Hyg breast cancer cells, (B) M13SV1-EGFP-Neo breast epithelial cells exhibiting stem cell characteristics, (C) M13HS-2 hybrid cells, (D) M13HS-8 hybrid cells. Statistical significance was calculated using Student's t-test: ***  =  p<0.001.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3646822&req=5

pone-0063711-g004: Knockdown of CCR7 expression abolish the CCL21 induced migration of M13HS hybrid cell lines.The migratory activity was analyzed using the 3D collagen matrix migration assay combined with time-lapse video-microscopy. For a better comparison the locomotory activities of CCL21 stimulated cells were calculated in relation to untreated control cells, which were set to 100%. Cells were stimulated with 500 ng/ml CCL21. Shown are the mean±SD of n = 3 independent experiments. (A) HS578T-Hyg breast cancer cells, (B) M13SV1-EGFP-Neo breast epithelial cells exhibiting stem cell characteristics, (C) M13HS-2 hybrid cells, (D) M13HS-8 hybrid cells. Statistical significance was calculated using Student's t-test: ***  =  p<0.001.
Mentions: Because the CCL21/CCR7 axis has been associated with lymph node metastasis of breast cancer (for review see: [34]) cell migration studies were conducted by using the 3D-collagen matrix migration assay. In addition to naïve cells cell migration studies were also performed with cells either transfected with specific CCR7 siRNA or scrambled control RNA (scRNA). CCR7 expression data are summarized in Figure 4A and show a reduced CCR7 expression in CCR7 siRNA transfected cells, whereas CCR7 levels of scRNA transfected cells were comparable to naïve cells (Figure 4A).

Bottom Line: Cell migration demonstrated that only M13HS hybrid cell lines, but not parental derivatives, responded to CCL21 stimulation with an increased migratory activity.Knockdown of CCR7 expression by siRNA completely abrogated the CCL21 induced migration of hybrid cell lines indicating the necessity of CCL21/CCR7 signaling.Because the CCL21/CCR7 axis has been linked to metastatic spreading of breast cancer to lymph nodes we conclude from our data that cell fusion could be a mechanism explaining the origin of metastatic cancer (hybrid) cells.

View Article: PubMed Central - PubMed

Affiliation: Institute of Immunology, Center for Biomedical Education and Research (ZBAF), Witten/Herdecke University, Witten, Germany.

ABSTRACT
The biological phenomenon of cell fusion has been linked to tumor progression because several data provided evidence that fusion of tumor cells and normal cells gave rise to hybrid cell lines exhibiting novel properties, such as increased metastatogenic capacity and an enhanced drug resistance. Here we investigated M13HS hybrid cell lines, derived from spontaneous fusion events between M13SV1-EGFP-Neo breast epithelial cells exhibiting stem cell characteristics and HS578T-Hyg breast cancer cells, concerning CCL21/CCR7 signaling. Western Blot analysis showed that all cell lines varied in their CCR7 expression levels as well as differed in the induction and kinetics of CCR7 specific signal transduction cascades. Flow cytometry-based calcium measurements revealed that a CCL21 induced calcium influx was solely detected in M13HS hybrid cell lines. Cell migration demonstrated that only M13HS hybrid cell lines, but not parental derivatives, responded to CCL21 stimulation with an increased migratory activity. Knockdown of CCR7 expression by siRNA completely abrogated the CCL21 induced migration of hybrid cell lines indicating the necessity of CCL21/CCR7 signaling. Because the CCL21/CCR7 axis has been linked to metastatic spreading of breast cancer to lymph nodes we conclude from our data that cell fusion could be a mechanism explaining the origin of metastatic cancer (hybrid) cells.

Show MeSH
Related in: MedlinePlus