Evaluation of protein kinase CK2 as a therapeutic target for squamous cell carcinoma of cats

Claire M. Cannon, Janeen H. Trembley, Betsy T. Kren, Gretchen M. Unger, M. Gerard O’Sullivan, Ingrid Cornax, Jaime F. Modiano, Khalil Ahmed

Research output: Contribution to journalArticle

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Abstract

OBJECTIVE To investigate protein kinase CK2 (CK2) expression in squamous cell carcinoma (SCC) of cats and to examine effects of CK2 downregulation on in vitro apoptosis and viability in SCC. SAMPLE Biopsy specimens of oral mucosa and testis and blood samples from clinically normal cats, biopsy specimens of oral SCC from cats, and feline SCC (SCCF1) and mammary gland carcinoma (K12) cell lines. PROCEDURES Immunohistochemical labeling for CK2α was performed on biopsy specimens. Sequences of the CK2α subunit gene and CK2α′ subunit gene in feline blood and feline cancer cell lines were determined by use of PCR and reverse-transcription PCR assays followed by direct Sanger sequencing. Specific small interfering RNAs (siRNAs) were developed for feline CK2α and CK2α′. The SCCF1 cells were treated with siRNA and assessed 72 hours later for CK2α and CK2α′ expression and markers of apoptosis (via western blot analysis) and for viability (via 3-[4,5-dimethylthiazol-2-yl]-5-[3-carboxymethoxyphenyl]-2-[4-sulfophenyl]-2H-tetrazolium assays). RESULTS CK2α was expressed in all feline oral mucosa samples and 7 of 8 oral SCC samples. Expression of CK2α and CK2α′ was successfully downregulated in SCCF1 cells by use of siRNAs, which resulted in decreased viability and induction of apoptosis. CONCLUSIONS AND CLINICAL RELEVANCE In this study, CK2 appeared to be a promising therapeutic target for SCCs of cats. A possible treatment strategy for SCCs of cats would be RNA interference that targets CK2.

Original languageEnglish (US)
Pages (from-to)946-955
Number of pages10
JournalAmerican journal of veterinary research
Volume78
Issue number8
DOIs
StatePublished - Aug 1 2017

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Casein Kinase II
Felidae
squamous cell carcinoma
protein kinases
Squamous Cell Carcinoma
Cats
cats
therapeutics
Small Interfering RNA
Mouth Mucosa
Apoptosis
Biopsy
Down-Regulation
small interfering RNA
digestive tract mucosa
Therapeutics
biopsy
Cell Line
Polymerase Chain Reaction
apoptosis

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Evaluation of protein kinase CK2 as a therapeutic target for squamous cell carcinoma of cats. / Cannon, Claire M.; Trembley, Janeen H.; Kren, Betsy T.; Unger, Gretchen M.; Gerard O’Sullivan, M.; Cornax, Ingrid; Modiano, Jaime F.; Ahmed, Khalil.

In: American journal of veterinary research, Vol. 78, No. 8, 01.08.2017, p. 946-955.

Research output: Contribution to journalArticle

Cannon, Claire M. ; Trembley, Janeen H. ; Kren, Betsy T. ; Unger, Gretchen M. ; Gerard O’Sullivan, M. ; Cornax, Ingrid ; Modiano, Jaime F. ; Ahmed, Khalil. / Evaluation of protein kinase CK2 as a therapeutic target for squamous cell carcinoma of cats. In: American journal of veterinary research. 2017 ; Vol. 78, No. 8. pp. 946-955.
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abstract = "OBJECTIVE To investigate protein kinase CK2 (CK2) expression in squamous cell carcinoma (SCC) of cats and to examine effects of CK2 downregulation on in vitro apoptosis and viability in SCC. SAMPLE Biopsy specimens of oral mucosa and testis and blood samples from clinically normal cats, biopsy specimens of oral SCC from cats, and feline SCC (SCCF1) and mammary gland carcinoma (K12) cell lines. PROCEDURES Immunohistochemical labeling for CK2α was performed on biopsy specimens. Sequences of the CK2α subunit gene and CK2α′ subunit gene in feline blood and feline cancer cell lines were determined by use of PCR and reverse-transcription PCR assays followed by direct Sanger sequencing. Specific small interfering RNAs (siRNAs) were developed for feline CK2α and CK2α′. The SCCF1 cells were treated with siRNA and assessed 72 hours later for CK2α and CK2α′ expression and markers of apoptosis (via western blot analysis) and for viability (via 3-[4,5-dimethylthiazol-2-yl]-5-[3-carboxymethoxyphenyl]-2-[4-sulfophenyl]-2H-tetrazolium assays). RESULTS CK2α was expressed in all feline oral mucosa samples and 7 of 8 oral SCC samples. Expression of CK2α and CK2α′ was successfully downregulated in SCCF1 cells by use of siRNAs, which resulted in decreased viability and induction of apoptosis. CONCLUSIONS AND CLINICAL RELEVANCE In this study, CK2 appeared to be a promising therapeutic target for SCCs of cats. A possible treatment strategy for SCCs of cats would be RNA interference that targets CK2.",
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AU - Cannon, Claire M.

AU - Trembley, Janeen H.

AU - Kren, Betsy T.

AU - Unger, Gretchen M.

AU - Gerard O’Sullivan, M.

AU - Cornax, Ingrid

AU - Modiano, Jaime F.

AU - Ahmed, Khalil

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N2 - OBJECTIVE To investigate protein kinase CK2 (CK2) expression in squamous cell carcinoma (SCC) of cats and to examine effects of CK2 downregulation on in vitro apoptosis and viability in SCC. SAMPLE Biopsy specimens of oral mucosa and testis and blood samples from clinically normal cats, biopsy specimens of oral SCC from cats, and feline SCC (SCCF1) and mammary gland carcinoma (K12) cell lines. PROCEDURES Immunohistochemical labeling for CK2α was performed on biopsy specimens. Sequences of the CK2α subunit gene and CK2α′ subunit gene in feline blood and feline cancer cell lines were determined by use of PCR and reverse-transcription PCR assays followed by direct Sanger sequencing. Specific small interfering RNAs (siRNAs) were developed for feline CK2α and CK2α′. The SCCF1 cells were treated with siRNA and assessed 72 hours later for CK2α and CK2α′ expression and markers of apoptosis (via western blot analysis) and for viability (via 3-[4,5-dimethylthiazol-2-yl]-5-[3-carboxymethoxyphenyl]-2-[4-sulfophenyl]-2H-tetrazolium assays). RESULTS CK2α was expressed in all feline oral mucosa samples and 7 of 8 oral SCC samples. Expression of CK2α and CK2α′ was successfully downregulated in SCCF1 cells by use of siRNAs, which resulted in decreased viability and induction of apoptosis. CONCLUSIONS AND CLINICAL RELEVANCE In this study, CK2 appeared to be a promising therapeutic target for SCCs of cats. A possible treatment strategy for SCCs of cats would be RNA interference that targets CK2.

AB - OBJECTIVE To investigate protein kinase CK2 (CK2) expression in squamous cell carcinoma (SCC) of cats and to examine effects of CK2 downregulation on in vitro apoptosis and viability in SCC. SAMPLE Biopsy specimens of oral mucosa and testis and blood samples from clinically normal cats, biopsy specimens of oral SCC from cats, and feline SCC (SCCF1) and mammary gland carcinoma (K12) cell lines. PROCEDURES Immunohistochemical labeling for CK2α was performed on biopsy specimens. Sequences of the CK2α subunit gene and CK2α′ subunit gene in feline blood and feline cancer cell lines were determined by use of PCR and reverse-transcription PCR assays followed by direct Sanger sequencing. Specific small interfering RNAs (siRNAs) were developed for feline CK2α and CK2α′. The SCCF1 cells were treated with siRNA and assessed 72 hours later for CK2α and CK2α′ expression and markers of apoptosis (via western blot analysis) and for viability (via 3-[4,5-dimethylthiazol-2-yl]-5-[3-carboxymethoxyphenyl]-2-[4-sulfophenyl]-2H-tetrazolium assays). RESULTS CK2α was expressed in all feline oral mucosa samples and 7 of 8 oral SCC samples. Expression of CK2α and CK2α′ was successfully downregulated in SCCF1 cells by use of siRNAs, which resulted in decreased viability and induction of apoptosis. CONCLUSIONS AND CLINICAL RELEVANCE In this study, CK2 appeared to be a promising therapeutic target for SCCs of cats. A possible treatment strategy for SCCs of cats would be RNA interference that targets CK2.

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