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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

Inhibition of CDK5 function through shRNA-mediated knockdown impairs in vivo tumorigenicity in orthotopic melanoma xenografts. (A) Growth of orthotopic SKMel-F10 xenografts was significantly inhibited upon induced CDK5 knockdown. (B) Sustained induced CDK5 knockdown was confirmed in explanted xenograft tumor tissues at the end of the experiment using Western blot analysis. Results from three representative doxycycline-induced and -uninduced mock-treated xenografts, respectively, are shown. (C) H&E stainings did not reveal any major discernible changes in the tumor microarchitecture upon induced CDK5 knockdown. Immunohistochemistry in harvested xenograft tumor tissues confirmed efficient knockdown of CDK5 at the protein level; proliferation as determined by Ki-67 immunolabeling was not significantly reduced after CDK5 depletion.
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f0020: Inhibition of CDK5 function through shRNA-mediated knockdown impairs in vivo tumorigenicity in orthotopic melanoma xenografts. (A) Growth of orthotopic SKMel-F10 xenografts was significantly inhibited upon induced CDK5 knockdown. (B) Sustained induced CDK5 knockdown was confirmed in explanted xenograft tumor tissues at the end of the experiment using Western blot analysis. Results from three representative doxycycline-induced and -uninduced mock-treated xenografts, respectively, are shown. (C) H&E stainings did not reveal any major discernible changes in the tumor microarchitecture upon induced CDK5 knockdown. Immunohistochemistry in harvested xenograft tumor tissues confirmed efficient knockdown of CDK5 at the protein level; proliferation as determined by Ki-67 immunolabeling was not significantly reduced after CDK5 depletion.

Mentions: To generate orthotopic xenografts, 2 × 106 SKMel-F10 melanoma cells were injected intradermally into the flanks of female NOD-SCID mice and allowed to grow for 2 weeks, at which point average tumor volumes reached approximately 80 mm3. Mice were then randomly assigned to either one of two study arms: The first group received doxycycline (200 μg/ml) and sucrose (2.5% w/v) in their drinking water continuously for the following 5 weeks, whereas the second group received only sucrose without addition of doxycycline. Of note, the group of mice in which CDK5 knockdown was induced (CDK5-F10 + dox) showed significantly inhibited tumor growth as compared with uninduced controls with preserved CDK5 expression (Figure 4A). Neither loss in body weight nor any other signs of distress or behavioral abnormalities were observed in mice from either of the two groups during the entire experiment. After 5 weeks, tumors were harvested, and persistent knockdown of CDK5 was confirmed by Western blot analysis (Figure 4B) as well as by immunohistochemistry (Figure 4C). Proliferation was assessed using Ki-67 labeling, and no discernible differences were found between the two arms, in line with in vitro data presented above. Also, no discernible histological differences were observed between xenografts derived from the CDK5 knockdown or control arm, respectively (Figure 4C).


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)

Inhibition of CDK5 function through shRNA-mediated knockdown impairs in vivo tumorigenicity in orthotopic melanoma xenografts. (A) Growth of orthotopic SKMel-F10 xenografts was significantly inhibited upon induced CDK5 knockdown. (B) Sustained induced CDK5 knockdown was confirmed in explanted xenograft tumor tissues at the end of the experiment using Western blot analysis. Results from three representative doxycycline-induced and -uninduced mock-treated xenografts, respectively, are shown. (C) H&E stainings did not reveal any major discernible changes in the tumor microarchitecture upon induced CDK5 knockdown. Immunohistochemistry in harvested xenograft tumor tissues confirmed efficient knockdown of CDK5 at the protein level; proliferation as determined by Ki-67 immunolabeling was not significantly reduced after CDK5 depletion.
© Copyright Policy - CC BY-NC-ND
Related In: Results  -  Collection

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

f0020: Inhibition of CDK5 function through shRNA-mediated knockdown impairs in vivo tumorigenicity in orthotopic melanoma xenografts. (A) Growth of orthotopic SKMel-F10 xenografts was significantly inhibited upon induced CDK5 knockdown. (B) Sustained induced CDK5 knockdown was confirmed in explanted xenograft tumor tissues at the end of the experiment using Western blot analysis. Results from three representative doxycycline-induced and -uninduced mock-treated xenografts, respectively, are shown. (C) H&E stainings did not reveal any major discernible changes in the tumor microarchitecture upon induced CDK5 knockdown. Immunohistochemistry in harvested xenograft tumor tissues confirmed efficient knockdown of CDK5 at the protein level; proliferation as determined by Ki-67 immunolabeling was not significantly reduced after CDK5 depletion.
Mentions: To generate orthotopic xenografts, 2 × 106 SKMel-F10 melanoma cells were injected intradermally into the flanks of female NOD-SCID mice and allowed to grow for 2 weeks, at which point average tumor volumes reached approximately 80 mm3. Mice were then randomly assigned to either one of two study arms: The first group received doxycycline (200 μg/ml) and sucrose (2.5% w/v) in their drinking water continuously for the following 5 weeks, whereas the second group received only sucrose without addition of doxycycline. Of note, the group of mice in which CDK5 knockdown was induced (CDK5-F10 + dox) showed significantly inhibited tumor growth as compared with uninduced controls with preserved CDK5 expression (Figure 4A). Neither loss in body weight nor any other signs of distress or behavioral abnormalities were observed in mice from either of the two groups during the entire experiment. After 5 weeks, tumors were harvested, and persistent knockdown of CDK5 was confirmed by Western blot analysis (Figure 4B) as well as by immunohistochemistry (Figure 4C). Proliferation was assessed using Ki-67 labeling, and no discernible differences were found between the two arms, in line with in vitro data presented above. Also, no discernible histological differences were observed between xenografts derived from the CDK5 knockdown or control arm, respectively (Figure 4C).

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