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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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CCR7 expression of parental cell lines and hybrid cell lines.CCR7 expression was detected by Western Blot analysis, whereby elf4E expression was used as an internal control. Shown are representative Western Blot data of n = 3 independent experiments.
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pone-0063711-g001: CCR7 expression of parental cell lines and hybrid cell lines.CCR7 expression was detected by Western Blot analysis, whereby elf4E expression was used as an internal control. Shown are representative Western Blot data of n = 3 independent experiments.

Mentions: M13HS-2 and M13HS-8 hybrid cell lines, which derived from spontaneous fusion events between M13SV1-EGFP-Neo human breast epithelial cells exhibiting stem cell characteristics and HS578T-Hyg human breast cancer cells, express the CCL21 receptor CCR7 (Figure 1). The CCR7 expression level of HS578T-Hyg breast cancer cells was similar to hybrid cell lines, whereas M13SV1-EGFP-Neo cells exhibited a weaker CCR7 expression (Figure 1). Because the interaction of CCL21 with its receptor CCR7 has been linked to lymph node metastasis of breast cancer and other tumor types (for review see: [34]), we first investigated whether hybrid cell lines respond to CCL21 with an activation of CCR7 specific signal transduction cascades including PI3K/AKT, MAPKp42/44 and PLC-β/γ signaling [27]. Suspension cells were stimulated for 1, 2, 5, and 10 minutes with 500 ng/ml CCL21 as well as Ly294002 (PI3K inhibitor; 500 nM) and PD98059 (MEK inhibitor; 500 nM) and subsequently analyzed for AKT and MAPKp42/44 phosphorylation (Figure 2). In addition to Figure 2 a densitometric analysis of Western Blot data has been performed, which is provided as supporting information (Figure S1).


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)

CCR7 expression of parental cell lines and hybrid cell lines.CCR7 expression was detected by Western Blot analysis, whereby elf4E expression was used as an internal control. Shown are representative Western Blot data of n = 3 independent experiments.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0063711-g001: CCR7 expression of parental cell lines and hybrid cell lines.CCR7 expression was detected by Western Blot analysis, whereby elf4E expression was used as an internal control. Shown are representative Western Blot data of n = 3 independent experiments.
Mentions: M13HS-2 and M13HS-8 hybrid cell lines, which derived from spontaneous fusion events between M13SV1-EGFP-Neo human breast epithelial cells exhibiting stem cell characteristics and HS578T-Hyg human breast cancer cells, express the CCL21 receptor CCR7 (Figure 1). The CCR7 expression level of HS578T-Hyg breast cancer cells was similar to hybrid cell lines, whereas M13SV1-EGFP-Neo cells exhibited a weaker CCR7 expression (Figure 1). Because the interaction of CCL21 with its receptor CCR7 has been linked to lymph node metastasis of breast cancer and other tumor types (for review see: [34]), we first investigated whether hybrid cell lines respond to CCL21 with an activation of CCR7 specific signal transduction cascades including PI3K/AKT, MAPKp42/44 and PLC-β/γ signaling [27]. Suspension cells were stimulated for 1, 2, 5, and 10 minutes with 500 ng/ml CCL21 as well as Ly294002 (PI3K inhibitor; 500 nM) and PD98059 (MEK inhibitor; 500 nM) and subsequently analyzed for AKT and MAPKp42/44 phosphorylation (Figure 2). In addition to Figure 2 a densitometric analysis of Western Blot data has been performed, which is provided as supporting information (Figure S1).

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