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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 knockdown prevents metastatic melanoma spread in murine dissemination models. (A) Formation of lung and liver metastases after intravenous injection of SKMel28 melanoma cells in immunodeficient HPRT NOD-SCID mice was determined. In this model system, doxycycline-induced shRNA-mediated CDK5 knockdown led to a dramatic reduction in the average number and size of pulmonary lesions as compared with uninduced controls (left panel). Liver metastases were completely abolished upon induced CDK5 knockdown, whereas they were readily observed in uninduced controls (right panel). The figures show representative H&E stains of FFPE tissue sections. (B) Injection of B16 murine melanoma cells into the tail veins of C57BL/6 wild-type mice was applied as a third, syngenic in vivo model system to validate the effect of CDK5 abrogation on melanoma metastasis. One of five CDK5-specific shRNA-clones (“CDK5#1”) showed reproducible knockdown of CDK5 in B16 murine melanoma cells as compared with scrambled controls using Western blot analysis. (C) When this clone CDK5#1 was used in a tail vein injection model, a dramatic decrease in both the amount and average size of lung colonies representing pulmonary metastases was observed upon CDK5 knockdown as compared to mock-transfected cells (macroscopic image from two representative animals).
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f0025: CDK5 knockdown prevents metastatic melanoma spread in murine dissemination models. (A) Formation of lung and liver metastases after intravenous injection of SKMel28 melanoma cells in immunodeficient HPRT NOD-SCID mice was determined. In this model system, doxycycline-induced shRNA-mediated CDK5 knockdown led to a dramatic reduction in the average number and size of pulmonary lesions as compared with uninduced controls (left panel). Liver metastases were completely abolished upon induced CDK5 knockdown, whereas they were readily observed in uninduced controls (right panel). The figures show representative H&E stains of FFPE tissue sections. (B) Injection of B16 murine melanoma cells into the tail veins of C57BL/6 wild-type mice was applied as a third, syngenic in vivo model system to validate the effect of CDK5 abrogation on melanoma metastasis. One of five CDK5-specific shRNA-clones (“CDK5#1”) showed reproducible knockdown of CDK5 in B16 murine melanoma cells as compared with scrambled controls using Western blot analysis. (C) When this clone CDK5#1 was used in a tail vein injection model, a dramatic decrease in both the amount and average size of lung colonies representing pulmonary metastases was observed upon CDK5 knockdown as compared to mock-transfected cells (macroscopic image from two representative animals).

Mentions: Next, we aimed to further evaluate these promising in vivo data using a complementary second in vivo model system, this time specifically mimicking the formation of systemic metastases. For this end, SKMel-F10 cells were injected into the tail veins of NOD-SCID mice. CDK5 knockdown was then induced in vivo by addition of doxycycline in drinking water, whereas uninduced mice served as controls. Interestingly, in this scenario, CDK5 blockade almost completely abrogated formation of lung colonies: uninduced SKMel-F10 cells formed multiple pulmonary lesions in all animals that were readily identified by gross inspection or histopathological examination, whereas doxycycline-induced CDK5-depleted melanoma cells yielded significantly fewer lesions of minimal size (Figure 5A). Again, these observations were reproducible in several sets of experiments and in line with in vitro data presented above.


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 knockdown prevents metastatic melanoma spread in murine dissemination models. (A) Formation of lung and liver metastases after intravenous injection of SKMel28 melanoma cells in immunodeficient HPRT NOD-SCID mice was determined. In this model system, doxycycline-induced shRNA-mediated CDK5 knockdown led to a dramatic reduction in the average number and size of pulmonary lesions as compared with uninduced controls (left panel). Liver metastases were completely abolished upon induced CDK5 knockdown, whereas they were readily observed in uninduced controls (right panel). The figures show representative H&E stains of FFPE tissue sections. (B) Injection of B16 murine melanoma cells into the tail veins of C57BL/6 wild-type mice was applied as a third, syngenic in vivo model system to validate the effect of CDK5 abrogation on melanoma metastasis. One of five CDK5-specific shRNA-clones (“CDK5#1”) showed reproducible knockdown of CDK5 in B16 murine melanoma cells as compared with scrambled controls using Western blot analysis. (C) When this clone CDK5#1 was used in a tail vein injection model, a dramatic decrease in both the amount and average size of lung colonies representing pulmonary metastases was observed upon CDK5 knockdown as compared to mock-transfected cells (macroscopic image from two representative animals).
© Copyright Policy - CC BY-NC-ND
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4562979&req=5

f0025: CDK5 knockdown prevents metastatic melanoma spread in murine dissemination models. (A) Formation of lung and liver metastases after intravenous injection of SKMel28 melanoma cells in immunodeficient HPRT NOD-SCID mice was determined. In this model system, doxycycline-induced shRNA-mediated CDK5 knockdown led to a dramatic reduction in the average number and size of pulmonary lesions as compared with uninduced controls (left panel). Liver metastases were completely abolished upon induced CDK5 knockdown, whereas they were readily observed in uninduced controls (right panel). The figures show representative H&E stains of FFPE tissue sections. (B) Injection of B16 murine melanoma cells into the tail veins of C57BL/6 wild-type mice was applied as a third, syngenic in vivo model system to validate the effect of CDK5 abrogation on melanoma metastasis. One of five CDK5-specific shRNA-clones (“CDK5#1”) showed reproducible knockdown of CDK5 in B16 murine melanoma cells as compared with scrambled controls using Western blot analysis. (C) When this clone CDK5#1 was used in a tail vein injection model, a dramatic decrease in both the amount and average size of lung colonies representing pulmonary metastases was observed upon CDK5 knockdown as compared to mock-transfected cells (macroscopic image from two representative animals).
Mentions: Next, we aimed to further evaluate these promising in vivo data using a complementary second in vivo model system, this time specifically mimicking the formation of systemic metastases. For this end, SKMel-F10 cells were injected into the tail veins of NOD-SCID mice. CDK5 knockdown was then induced in vivo by addition of doxycycline in drinking water, whereas uninduced mice served as controls. Interestingly, in this scenario, CDK5 blockade almost completely abrogated formation of lung colonies: uninduced SKMel-F10 cells formed multiple pulmonary lesions in all animals that were readily identified by gross inspection or histopathological examination, whereas doxycycline-induced CDK5-depleted melanoma cells yielded significantly fewer lesions of minimal size (Figure 5A). Again, these observations were reproducible in several sets of experiments and in line with in vitro data presented above.

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