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Cyclin-Dependent Kinase 5 (CDK5) Controls Melanoma Cell Motility, Invasiveness, and Metastatic Spread-Identification of a Promising Novel therapeutic target.

Bisht S, Nolting J, Schütte U, Haarmann J, Jain P, Shah D, Brossart P, Flaherty P, Feldmann G - Transl Oncol (2015)

Bottom Line: In vivo, CDK5 knockdown inhibited growth of orthotopic xenografts as well as formation of lung and liver colonies in xenogenic injection models mimicking systemic metastases.CDK5 knockdown was accompanied by dephosphorylation and overexpression of caldesmon, and concomitant caldesmon knockdown rescued cell motility and proinvasive phenotype.Finally, it was found that pharmacological inhibition of CDK5 activity by means of roscovitine as well as by a novel small molecule CDK5-inhibitor, N-(5-isopropylthiazol-2-yl)-3-phenylpropanamide, similarly caused marked inhibition of invasion/migration, colony formation, and anchorage-independent growth of melanoma cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Internal Medicine 3, Center of Integrated Oncology (CIO) Cologne-Bonn, University Hospital of Bonn, Germany.

No MeSH data available.


Related in: MedlinePlus

CDK5 affects melanoma invasiveness and migration through regulation of caldesmon. (A) shRNA-mediated knockdown of CDK5 causes upregulation and dephosphorylation of caldesmon in SKMel-F10 and SKMel-B7 cells in the presence of doxycycline as compared with uninduced or mock-transduced cells as demonstrated using Western blot analysis. (B) Successful siRNA-mediated knockdown of caldesmon is demonstrated in CDK5 knockdown cells. Note that CDK5 protein levels are not affected by caldesmon knockdown. (C) Caldesmon knockdown rescues invasiveness and migration of CDK5-depleted SKMel human melanoma cells. The upper panel shows membranes from modified Boyden chamber assays; loss of invasion/migration upon doxycycline-induced CDK5 knockdown is rescued by concomitant siRNA-mediated knockdown of caldesmon, whereas no effect is found in scrambled siRNA controls. Relative cell counts as means and standard deviations are shown in the lower panel. *P < .05. (D) In wound-healing assays, concomitant caldesmon knockdown by means of siRNA transfection leads to increased cell motility, thus rescuing the observed reduction in wound healing upon downregulation of CDK5 as shown above.
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f0030: CDK5 affects melanoma invasiveness and migration through regulation of caldesmon. (A) shRNA-mediated knockdown of CDK5 causes upregulation and dephosphorylation of caldesmon in SKMel-F10 and SKMel-B7 cells in the presence of doxycycline as compared with uninduced or mock-transduced cells as demonstrated using Western blot analysis. (B) Successful siRNA-mediated knockdown of caldesmon is demonstrated in CDK5 knockdown cells. Note that CDK5 protein levels are not affected by caldesmon knockdown. (C) Caldesmon knockdown rescues invasiveness and migration of CDK5-depleted SKMel human melanoma cells. The upper panel shows membranes from modified Boyden chamber assays; loss of invasion/migration upon doxycycline-induced CDK5 knockdown is rescued by concomitant siRNA-mediated knockdown of caldesmon, whereas no effect is found in scrambled siRNA controls. Relative cell counts as means and standard deviations are shown in the lower panel. *P < .05. (D) In wound-healing assays, concomitant caldesmon knockdown by means of siRNA transfection leads to increased cell motility, thus rescuing the observed reduction in wound healing upon downregulation of CDK5 as shown above.

Mentions: Of interest, it was found that doxycycline-induced depletion of CDK5 in either SKMel-F10 or -B7 cells led to dephosphorylation of caldesmon while at the same time causing an increase of total caldesmon as observed using Western blot analysis. No change in phosphorylation status was found in uninduced SKMel-F10 or -B7 cells or in mock-transduced controls (Figure 6A).


Cyclin-Dependent Kinase 5 (CDK5) Controls Melanoma Cell Motility, Invasiveness, and Metastatic Spread-Identification of a Promising Novel therapeutic target.

Bisht S, Nolting J, Schütte U, Haarmann J, Jain P, Shah D, Brossart P, Flaherty P, Feldmann G - Transl Oncol (2015)

CDK5 affects melanoma invasiveness and migration through regulation of caldesmon. (A) shRNA-mediated knockdown of CDK5 causes upregulation and dephosphorylation of caldesmon in SKMel-F10 and SKMel-B7 cells in the presence of doxycycline as compared with uninduced or mock-transduced cells as demonstrated using Western blot analysis. (B) Successful siRNA-mediated knockdown of caldesmon is demonstrated in CDK5 knockdown cells. Note that CDK5 protein levels are not affected by caldesmon knockdown. (C) Caldesmon knockdown rescues invasiveness and migration of CDK5-depleted SKMel human melanoma cells. The upper panel shows membranes from modified Boyden chamber assays; loss of invasion/migration upon doxycycline-induced CDK5 knockdown is rescued by concomitant siRNA-mediated knockdown of caldesmon, whereas no effect is found in scrambled siRNA controls. Relative cell counts as means and standard deviations are shown in the lower panel. *P < .05. (D) In wound-healing assays, concomitant caldesmon knockdown by means of siRNA transfection leads to increased cell motility, thus rescuing the observed reduction in wound healing upon downregulation of CDK5 as shown above.
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f0030: CDK5 affects melanoma invasiveness and migration through regulation of caldesmon. (A) shRNA-mediated knockdown of CDK5 causes upregulation and dephosphorylation of caldesmon in SKMel-F10 and SKMel-B7 cells in the presence of doxycycline as compared with uninduced or mock-transduced cells as demonstrated using Western blot analysis. (B) Successful siRNA-mediated knockdown of caldesmon is demonstrated in CDK5 knockdown cells. Note that CDK5 protein levels are not affected by caldesmon knockdown. (C) Caldesmon knockdown rescues invasiveness and migration of CDK5-depleted SKMel human melanoma cells. The upper panel shows membranes from modified Boyden chamber assays; loss of invasion/migration upon doxycycline-induced CDK5 knockdown is rescued by concomitant siRNA-mediated knockdown of caldesmon, whereas no effect is found in scrambled siRNA controls. Relative cell counts as means and standard deviations are shown in the lower panel. *P < .05. (D) In wound-healing assays, concomitant caldesmon knockdown by means of siRNA transfection leads to increased cell motility, thus rescuing the observed reduction in wound healing upon downregulation of CDK5 as shown above.
Mentions: Of interest, it was found that doxycycline-induced depletion of CDK5 in either SKMel-F10 or -B7 cells led to dephosphorylation of caldesmon while at the same time causing an increase of total caldesmon as observed using Western blot analysis. No change in phosphorylation status was found in uninduced SKMel-F10 or -B7 cells or in mock-transduced controls (Figure 6A).

Bottom Line: In vivo, CDK5 knockdown inhibited growth of orthotopic xenografts as well as formation of lung and liver colonies in xenogenic injection models mimicking systemic metastases.CDK5 knockdown was accompanied by dephosphorylation and overexpression of caldesmon, and concomitant caldesmon knockdown rescued cell motility and proinvasive phenotype.Finally, it was found that pharmacological inhibition of CDK5 activity by means of roscovitine as well as by a novel small molecule CDK5-inhibitor, N-(5-isopropylthiazol-2-yl)-3-phenylpropanamide, similarly caused marked inhibition of invasion/migration, colony formation, and anchorage-independent growth of melanoma cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Internal Medicine 3, Center of Integrated Oncology (CIO) Cologne-Bonn, University Hospital of Bonn, Germany.

No MeSH data available.


Related in: MedlinePlus