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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

Expression of CDK5 and its activator protein CDK5R1/p35 in human melanoma tissues and cell lines. (A) CDK5R1/p35 was found to be overexpressed in primary melanoma as well as in metastatic lesions as compared with normal skin using immunohistochemistry. (B) In tissue microarrays, intermediate (2 +) or high (3 +) p35 immunolabeling was more commonly observed in malignant melanoma tissue as compared with benign skin. (C) Western blot analysis of CDK5, p35, and p25 steady-state expression levels in five human melanoma cell lines.
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f0005: Expression of CDK5 and its activator protein CDK5R1/p35 in human melanoma tissues and cell lines. (A) CDK5R1/p35 was found to be overexpressed in primary melanoma as well as in metastatic lesions as compared with normal skin using immunohistochemistry. (B) In tissue microarrays, intermediate (2 +) or high (3 +) p35 immunolabeling was more commonly observed in malignant melanoma tissue as compared with benign skin. (C) Western blot analysis of CDK5, p35, and p25 steady-state expression levels in five human melanoma cell lines.

Mentions: CDK5 has been described as ubiquitously expressed in mammalian tissues including melanoma cells [19], with its functional activity being regulated by and strictly dependent on its association with either one of its two activator proteins, CDK5R1/p35 or CDK5R2/p39, respectively. In other words, expression of either CDK5R1/p35 or CDK5R2/p39 is equivalent to CDK5 activation throughout various tissues [9,20–22]. Therefore, we evaluated p35 expression in human melanoma cells and primary tissue samples. Firstly, tissue microarrays (US Biomax, Inc., Rockville, MD) representing 86 cases of primary malignant melanoma lesions, 50 samples of melanoma metastases, and 27 control samples of normal skin tissue, with up to 3 replicates for each of the total 163 cases, were stained for p35 expression using immunohistochemistry. Each sample was scored as 0 (negative), 1 +, 2 +, or 3 + based on cytoplasmatic p35 immunolabeling; in cases with different values among replicate samples, the highest value was counted. Overexpression of p35 as compared with normal skin samples was found in 70% of melanoma tissues tested, including both primary and metastatic lesions. Thirty-six percent (49/136 cases) of melanomas (34 of 86 primary and 15 out of 50 metastatic lesions, respectively) showed high protein expression scored as “3+” as compared with basal immunolabeling (0 to 1 +) observed in 100% (27 of 27 informative cases) of normal skin tissues. Intermediate (scored as “2+”) expression was found in 34% (46/136 cases) of melanoma samples (31/86 primary and 15/50 metastatic melanoma lesions, respectively) but in none of the non-neoplastic normal skin control samples (Figure 1, A and B).


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)

Expression of CDK5 and its activator protein CDK5R1/p35 in human melanoma tissues and cell lines. (A) CDK5R1/p35 was found to be overexpressed in primary melanoma as well as in metastatic lesions as compared with normal skin using immunohistochemistry. (B) In tissue microarrays, intermediate (2 +) or high (3 +) p35 immunolabeling was more commonly observed in malignant melanoma tissue as compared with benign skin. (C) Western blot analysis of CDK5, p35, and p25 steady-state expression levels in five human melanoma cell lines.
© Copyright Policy - CC BY-NC-ND
Related In: Results  -  Collection

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

f0005: Expression of CDK5 and its activator protein CDK5R1/p35 in human melanoma tissues and cell lines. (A) CDK5R1/p35 was found to be overexpressed in primary melanoma as well as in metastatic lesions as compared with normal skin using immunohistochemistry. (B) In tissue microarrays, intermediate (2 +) or high (3 +) p35 immunolabeling was more commonly observed in malignant melanoma tissue as compared with benign skin. (C) Western blot analysis of CDK5, p35, and p25 steady-state expression levels in five human melanoma cell lines.
Mentions: CDK5 has been described as ubiquitously expressed in mammalian tissues including melanoma cells [19], with its functional activity being regulated by and strictly dependent on its association with either one of its two activator proteins, CDK5R1/p35 or CDK5R2/p39, respectively. In other words, expression of either CDK5R1/p35 or CDK5R2/p39 is equivalent to CDK5 activation throughout various tissues [9,20–22]. Therefore, we evaluated p35 expression in human melanoma cells and primary tissue samples. Firstly, tissue microarrays (US Biomax, Inc., Rockville, MD) representing 86 cases of primary malignant melanoma lesions, 50 samples of melanoma metastases, and 27 control samples of normal skin tissue, with up to 3 replicates for each of the total 163 cases, were stained for p35 expression using immunohistochemistry. Each sample was scored as 0 (negative), 1 +, 2 +, or 3 + based on cytoplasmatic p35 immunolabeling; in cases with different values among replicate samples, the highest value was counted. Overexpression of p35 as compared with normal skin samples was found in 70% of melanoma tissues tested, including both primary and metastatic lesions. Thirty-six percent (49/136 cases) of melanomas (34 of 86 primary and 15 out of 50 metastatic lesions, respectively) showed high protein expression scored as “3+” as compared with basal immunolabeling (0 to 1 +) observed in 100% (27 of 27 informative cases) of normal skin tissues. Intermediate (scored as “2+”) expression was found in 34% (46/136 cases) of melanoma samples (31/86 primary and 15/50 metastatic melanoma lesions, respectively) but in none of the non-neoplastic normal skin control samples (Figure 1, A and B).

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