casein-kinase-ii and Carcinoma--Squamous-Cell

Protein kinase CK2 (previously known as casein kinase II) is a protein serine/threonine kinase that has been implicated in cell growth and proliferation. The focus of this review is on the apparent role of CK2 in cancer. Studies from several laboratories have shown a dysregulated expression of the kinase in tumors. Nuclear matrix and chromatin appear to be key sites for signaling of the CK2 activity in relation to cell growth. Several types of growth stimuli produce a common downstream response in CK2 by enhancing its nuclear shuttling. The neoplastic change is also associated with changes in intracellular localization of the kinase so that a higher nuclear localization is observed in tumor cells compared with normal cells. Experimental studies suggest that dysregulated expression of the alpha subunit of CK2 imparts an oncogenic potential in the cells such that in cooperation with certain oncogenes it produces a profound enhancement of the tumor phenotype. Recent studies have provided evidence that overexpression of CK2 in tumor cells is not simply a reflection of tumor cell proliferation alone but additionally may reflect the pathobiological characteristics of the tumor. Of considerable interest is the possibility that CK2 dysregulation in tumors may influence the apoptotic activity in those cells. Approaches to interfering with the CK2 signal may provide a useful means for inducing tumor cell death.

Topics: Adenocarcinoma; Animals; Apoptosis; Carcinoma, Squamous Cell; Casein Kinase II; Cell Nucleus; Chromatin; Head and Neck Neoplasms; Humans; Immunohistochemistry; Male; Neoplasms; Nuclear Matrix; Prostatic Neoplasms; Protein Serine-Threonine Kinases; Signal Transduction

Protein kinase CK2 (CK2) is a highly promising target for cancer therapy, and anti-CK2 gene expression therapy has shown effectiveness in rodent models of human head and neck cancer (HNC). To date, there has been no large-animal model of cancer in which to further explore anti-CK2 therapies. Feline oral squamous cell carcinoma (FOSCC) has been proposed as a large-animal model for human HNC, and we have previously shown that CK2 is a rational target in FOSCC. Here we have tested the hypothesis that a novel tenfibgen-coated tumor-specific nanocapsule carrying RNA interference (RNAi) oligonucleotides targeting feline CK2α and CK2α' (TBG-RNAi-fCK2αα') would be safe in cats with FOSCC; assessment of target inhibition and tumor response were secondary aims. Nine cats were enrolled and treated at two dose levels in a 3+3 escalation. Cats received a total of six treatments with TBG-RNAi-fCK2αα'. Pre- and posttreatment, tumor and normal oral mucosa biopsies were collected to assess CK2 expression, using immunohistochemistry (IHC) preparations evaluated by light microscopy. Toxicity and tumor response were assessed on the basis of standard criteria. The most common adverse events were grade 1 or 2 weight loss and anorexia. Grade 3 tissue necrosis was seen in association with tumor response in one cat, asymptomatic grade 4 elevations in aspartate transaminase and creatine phosphokinase in one cat, and asymptomatic grade 3 hypokalemia in one cat. Of six cats with evaluable biopsies, two had a reduction in CK2 IHC score in tumors after treatment. Four cats had progressive disease during the study period, three had stable disease, one had partial response, and response could not be evaluated in one cat. We conclude that the drug appeared safe and that there is some evidence of efficacy in FOSCC. Further investigation regarding dosing, schedule, target modulation, toxicity, and efficacy in a larger group of cats is warranted and may inform future clinical studies in human head and neck cancer.

Topics: Animals; Anorexia; Carcinoma, Squamous Cell; Casein Kinase II; Cats; Cells, Cultured; Female; Humans; Hypokalemia; Male; Mouth Mucosa; Mouth Neoplasms; RNA, Small Interfering; RNAi Therapeutics; Weight Loss

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.

Topics: Animals; Antineoplastic Agents; Apoptosis; Blotting, Western; Carcinoma, Squamous Cell; Casein Kinase II; Cat Diseases; Cats; Cell Line; Down-Regulation; Drug Delivery Systems; Male; Mouth Mucosa; RNA, Small Interfering; Testis

N-α-acetyltransferase D (NatD) mediates N-α-terminal acetylation (Nt-acetylation) of histone H4 known to be involved in cell growth. Here we report that NatD promotes the migratory and invasive capabilities of lung cancer cells in vitro and in vivo. Depletion of NatD suppresses the epithelial-to-mesenchymal transition (EMT) of lung cancer cells by directly repressing the expression of transcription factor Slug, a key regulator of EMT. We found that Nt-acetylation of histone H4 antagonizes histone H4 serine 1 phosphorylation (H4S1ph), and that downregulation of Nt-acetylation of histone H4 facilitates CK2α binding to histone H4 in lung cancer cells, resulting in increased H4S1ph and epigenetic reprogramming to suppress Slug transcription to inhibit EMT. Importantly, NatD is commonly upregulated in primary human lung cancer tissues where its expression level correlates with Slug expression, enhanced invasiveness, and poor clinical outcomes. These findings indicate that NatD is a crucial epigenetic modulator of cell invasion during lung cancer progression.NatD is an acetyltransferase responsible for N-α-terminal acetylation of the histone H4 and H2A and has been linked to cell growth. Here the authors show that NatD-mediated acetylation of histone H4 serine 1 competes with the phosphorylation by CK2α at the same residue thus leading to the upregulation of Slug and tumor progression.

Topics: A549 Cells; Adenocarcinoma; Animals; Carcinoma, Squamous Cell; Casein Kinase II; Cell Movement; China; Epithelial-Mesenchymal Transition; Female; HEK293 Cells; Histones; Humans; Lung Neoplasms; Male; Mice; Middle Aged; N-Terminal Acetyltransferase D; Neoplasm Invasiveness; Phosphorylation; Snail Family Transcription Factors

Improved survival for patients with head and neck cancers (HNC) with recurrent and metastatic disease warrants that cancer therapy is specific, with protected delivery of the therapeutic agent to primary and metastatic cancer cells. A further objective should be that downregulation of the intracellular therapy target leads to cell death without compensation by an alternate pathway. To address these goals, we report the utilization of a sub-50-nm tenfibgen (s50-TBG) nanocapsule that delivers RNAi oligonucleotides directed against the essential survival signal protein kinase CK2 (RNAi-CK2) in a cancer cell-specific manner. We have evaluated mechanism and efficacy of using s50-TBG-RNAi-CK2 nanocapsules for therapy of primary and metastatic head and neck squamous cell carcinoma (HNSCC). s50-TBG nanocapsules enter cancer cells via the lipid raft/caveolar pathway and deliver their cargo (RNAi-CK2) preferentially to malignant but not normal tissues in mice. Our data suggest that RNAi-CK2, a unique single-stranded oligonucleotide, co-opts the argonaute 2/RNA-induced silencing complex pathway to target the CK2αα' mRNAs. s50-TBG-RNAi-CK2 inhibited cell growth corresponding with reduced CK2 expression in targeted tumor cells. Treatment of three xenograft HNSCC models showed that primary tumors and metastases responded to s50-TBG-RNAi-CK2 therapy, with tumor shrinkage and 6-month host survival that was achieved at relatively low doses of the therapeutic agent without any adverse toxic effect in normal tissues in the mice. We suggest that our nanocapsule technology and anti-CK2 targeting combine into a therapeutic modality with a potential of significant translational promise.

Topics: Animals; Argonaute Proteins; Base Sequence; Carcinoma, Squamous Cell; Casein Kinase II; Female; Gene Knockdown Techniques; Head and Neck Neoplasms; Humans; Membrane Microdomains; Mice, Inbred BALB C; Mice, Nude; Nanocapsules; Neoplasm Transplantation; Peptide Fragments; RNA Interference; RNA, Small Interfering; Splenic Neoplasms; Tenascin; Tissue Distribution; Transfection

Cancer stem cells (CSC) and genes have been linked to cancer development and therapeutic resistance, but the signaling mechanisms regulating CSC genes and phenotype are incompletely understood. CK2 has emerged as a key signal serine/threonine kinase that modulates diverse signal cascades regulating cell fate and growth. We previously showed that CK2 is often aberrantly expressed and activated in head and neck squamous cell carcinomas (HNSCC), concomitantly with mutant (mt) tumor suppressor TP53, and inactivation of its family member, TAp73. Unexpectedly, we observed that classical stem cell genes Nanog, Sox2, and Oct4, are overexpressed in HNSCC with inactivated TAp73 and mtTP53. However, the potential relationship between CK2, TAp73 inactivation, and CSC phenotype is unknown. We reveal that inhibition of CK2 by pharmacologic inhibitors or siRNA inhibits the expression of CSC genes and side population (SP), while enhancing TAp73 mRNA and protein expression. Conversely, CK2 inhibitor attenuation of CSC protein expression and the SP by was abrogated by TAp73 siRNA. Bioinformatic analysis uncovered a single predicted CK2 threonine phosphorylation site (T27) within the N-terminal transactivation domain of TAp73. Nuclear CK2 and TAp73 interaction, confirmed by co-immunoprecipitation, was attenuated by CK2 inhibitor, or a T27A point-mutation of this predicted CK2 threonine phospho-acceptor site of TAp73. Further, T27A mutation attenuated phosphorylation, while enhancing TAp73 function in repressing CSC gene expression and SP cells. A new CK2 inhibitor, CX-4945, inhibited CSC related SP cells, clonogenic survival, and spheroid formation. Our study unveils a novel regulatory mechanism whereby aberrant CK2 signaling inhibits TAp73 to promote the expression of CSC genes and phenotype.

Topics: Benzimidazoles; Carcinoma, Squamous Cell; Casein Kinase II; Cell Line, Tumor; DNA-Binding Proteins; Enzyme Inhibitors; Gene Expression Regulation, Neoplastic; Head and Neck Neoplasms; Homeodomain Proteins; Humans; Mutation; Nanog Homeobox Protein; Neoplastic Stem Cells; Nuclear Proteins; Octamer Transcription Factor-3; Phosphorylation; RNA, Small Interfering; SOXB1 Transcription Factors; Squamous Cell Carcinoma of Head and Neck; Threonine; Tumor Protein p73; Tumor Suppressor Protein p53; Tumor Suppressor Proteins

To investigate the expression of casein kinase 2β in esophageal carcinoma tissues and analyze its clinical significance.. The expression of CK2β in a tissue chip containing 60 normal esophageal mucosa and 60 colorectal cancer specimens were detected immunohistochemically. Ten human esophageal carcinoma and adjacent normal tissues were examined for the expression of CK2β protein and mRNA using Western blotting and real-time quantitative PCR, respectively.. A strong expression of CK2β was found in 85.71% of the esophageal cancer tissues; 1.79% of the cancer tissues were negative for CK2β expression, and 1.79% and 10.71% of the cancer tissues were weakly and moderately positive, respectively. In the normal mucosal tissues, 96.67% of the tissues were negative for CK2β and 3.33% showed weak CK2β expression, showing a significant difference in the distribution of CK2β between normal and esophageal carcinoma tissues (P<0.001). The expression level of CK2β in esophageal cancers was associated with the pathological stage (TNM) (P=0.010). Western blotting and real-time quantitative PCR also confirmed an increased CK2β expression in the esophageal cancer tissues.. The high expression of protein kinase CK2β is closely related to the carcinogenesis and malignancy of esophageal cancer.

Topics: Adult; Aged; Carcinoma, Squamous Cell; Casein Kinase II; Esophageal Neoplasms; Female; Gene Expression Regulation, Neoplastic; Humans; Male; Middle Aged; Neoplasm Staging

Squamous cell carcinoma of the head and neck (SCCHN) is the seventh most common cancer worldwide. Unfortunately, the survival of patients with SCCHN has not improved in the last 40 years, and thus new targets for therapy are needed. Recently, elevations in serum level of interleukin 6 (IL-6) and expression of Twist in tumor samples were found to be associated with poor clinical outcomes in multiple types of cancer, including SCCHN. Although Twist has been proposed as a master regulator of epithelial-mesenchymal transition and metastasis in cancers, the mechanisms by which Twist levels are regulated post-translationally are not completely understood. Tumor progression is characterized by the involvement of cytokines and growth factors and Twist induction has been connected with a number of these signaling pathways including IL-6. Since many of the effects of IL-6 are mediated through activation of protein phosphorylation cascades, this implies that Twist expression must be under a tight control at the post-translational level in order to respond in a timely manner to external stimuli.. Our data show that IL-6 increases Twist expression via a transcription-independent mechanism in many SCCHN cell lines. Further investigation revealed that IL-6 stabilizes Twist in SCCHN cell lines through casein kinase 2 (CK2) phosphorylation of Twist residues S18 and S20, and that this phosphorylation inhibits degradation of Twist. Twist phosphorylation not only increases its stability but also enhances cell motility. Thus, post-translational modulation of Twist contributes to its tumor-promoting properties.. Our study shows Twist expression can be regulated at the post-translational level through phosphorylation by CK2, which increases Twist stability in response to IL-6 stimulation. Our findings not only provide novel mechanistic insights into post-translational regulation of Twist but also suggest that CK2 may be a viable therapeutic target in SCCHN.

Topics: Carcinoma, Squamous Cell; Casein Kinase II; Cell Line, Tumor; Enzyme Activation; Gene Expression Regulation, Enzymologic; Gene Expression Regulation, Neoplastic; Head and Neck Neoplasms; Humans; Interleukin-6; Neoplasm Metastasis; Nuclear Proteins; Phosphorylation; Protein Processing, Post-Translational; Reverse Transcriptase Polymerase Chain Reaction; Twist-Related Protein 1; Wound Healing

The aim of this study is to investigate the expression of CK2 subunits and CK2 effects on NF-kappaB-mediated and TP53-mediated signal activation and gene expression, the malignant phenotype, and chemosensitivity in head and neck squamous cell carcinoma (HNSCC) in vitro and in vivo.. Protein expression of CK2 subunits was investigated by Western blot and immunohistochemistry. CK2 subunits were knocked down by small interfering RNA, and NF-kappaB activation was examined using DNA binding, Western blot, and luciferase reporter assays. Gene expression was measured by quantitative reverse transcription-PCR. Cell growth, survival, motility, and sensitivity to cisplatin were measured by MTT, flow cytometry, and migration assays. In vivo targeting of CK2alpha/alpha' in HNSCC xenograft models was achieved using anti-CK2alpha/alpha' oligodeoxynucleotide encapsulated in sub-50-nm tenfibgen nanocapsules.. CK2 subunit proteins were overexpressed in HNSCC lines and tissues. Knockdown of CK2 subunits differentially inhibited IkappaBalpha degradation, NF-kappaB nuclear localization, phosphorylation, DNA binding, and reporter activity. CK2 subunits modulated gene expression and the malignant phenotype involved in cell cycle and migration, whereas CK2alpha is critical to promote proliferation, antiapoptosis, and cisplatin resistance in vitro. Furthermore, in vivo delivery of anti-CK2alpha/alpha' oligodeoxynucleotide nanocapsules significantly suppressed tumor growth in HNSCC xenograft models, in association with modulation of CK2 and NF-kappaB regulated molecules, TP53 family proteins, and induction of apoptosis.. Our study reveals a novel role of CK2 in coregulating NF-kappaB activation, TP53/p63 expression, and downstream gene expression. Downregulation of CK2 in HNSCC models in vitro and in vivo shows antitumor effects as well as sensitization to cisplatin.

Topics: Animals; Apoptosis; Blotting, Western; Carcinoma, Squamous Cell; Casein Kinase II; Cell Cycle; Cell Movement; Cell Proliferation; Head and Neck Neoplasms; Humans; Luciferases; Male; Mice; Mice, Inbred BALB C; Mice, Nude; Nanocapsules; NF-kappa B; Oligonucleotides, Antisense; Phenotype; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; RNA, Small Interfering; Tumor Cells, Cultured; Tumor Suppressor Protein p53; Wound Healing

To investigate the expression of protein kinase CK2 and its relationship with the development, progress, invasion and metastasis of squamous cell carcinoma of larynx (LSCC).. Immunohistochemical SP staining method was used to assess the expression of CK2 in 18 cases of normal laryngeal mucosa, 14 cases of polyp of vocal cord, 11 cases of larynx papilloma and 50 cases of LSCC patients. And RT-PCR was used to detect the expression of CK2alpha mRNA and CK2beta mRNA in 50 cases of LSCG patients. The relationship between CK2alpha and CK2p was evaluated.. The positive expression rate of CK2alpha and CK2beta in normal laryngeal mucosa, polyp of vocal cord and tissues close to carcinoma by 1.0 cm, nonmetastatic lymph nodes were lower than that in tissues close to carcinoma by 0.5 cm and laryngeal papilloma. The positive expression rate of CK2alpha and CK2beta in laryngeal carcinoma and metastatic lymph nodes were the highest among the groups. The expression rate of CK2alpha and CK2beta in tissues of laryngeal carcinoma and metastatic lymph nodes of neck was significantly higher than that of laryngeal papilloma and tissues close to carcinoma by 0.5 cm (P < 0.05). In the group of LSCC, the expression of CK2alpha in G2 and in G3 was significantly higher than that in G1 (P < 0.05). While the age of the patients, TNM stage and lymphatic metastasis didn't change in the expression of CK2alpha obviously. The expression of CK2beta correlates to the differentiation grading and lymphatic metastasis in LSCC patients, but not to the age and TNM stage. The result from RT-PCR was highly consistent with that from immunohistochemical SP staining. There was a positive correlation between the expression of CK2alpha in LSCC patients and that of CK2beta.. The over expression of protein kinase CK2 may be an accelerator to the formation and development of LSCC. Protein kinase CK2 may be one of the predictors for the malignant grade of LSCC. To inhibit the over expression might be new therapeutic methods for LSCC.

Topics: Adult; Aged; Carcinoma, Squamous Cell; Casein Kinase II; Female; Humans; Laryngeal Mucosa; Laryngeal Neoplasms; Male; Middle Aged; Neoplasm Staging; Precancerous Conditions

We showed previously that the signal transcription factor nuclear factor-kappaB (NF-kappaB) is aberrantly activated and that inhibition of NF-kappaB induces cell death and inhibits tumorigenesis in head and neck squamous cell carcinomas (HNSCC). Thus, identification of specific kinases underlying the activation of NF-kappaB could provide targets for selective therapy. Inhibitor-kappaB (IkappaB) kinase (IKK) is known to activate NF-kappaB by inducing NH(2)-terminal phosphorylation and degradation of its endogenous inhibitor, IkappaB. Casein kinase 2 (CK2) was previously reported to be overexpressed in HNSCC cells and to be a COOH-terminal IKK, but its relationship to NF-kappaB activation in HNSCC cells is unknown. In this study, we examined the contribution of IKK and CK2 in the regulation of NF-kappaB in HNSCC in vitro. NF-kappaB activation was specifically inhibited by kinase-dead mutants of the IKK1 and IKK2 subunits or small interfering RNA targeting the beta subunit of CK2. CK2 and IKK kinase activity, as well as NF-kappaB transcriptional activity, was shown to be serum responsive, indicating that these kinases mediate aberrant activation of NF-kappaB in response to serum factor(s) in vitro. Recombinant CK2alpha was shown to phosphorylate recombinant IKK2 as well as to promote immunoprecipitated IKK complex from HNSCC to phosphorylate the NH(2)-terminal S32/S36 of IkappaBalpha. We conclude that the aberrant NF-kappaB activity in HNSCC cells in response to serum is partially through a novel mechanism involving CK2-mediated activation of IKK2, making these kinases candidates for selective therapy to target the NF-kappaB pathway in HNSCC.

Topics: Carcinoma, Squamous Cell; Casein Kinase II; Cell Line, Tumor; Culture Media, Serum-Free; Head and Neck Neoplasms; Humans; I-kappa B Kinase; I-kappa B Proteins; NF-kappa B; Phosphorylation; RNA, Small Interfering; Serum; Transfection

To investigate the expression of Protein kinase CK2alpha in squamous cell carcinoma of larynx (LSCC) and to evaluate its clinical significance.. The expressions of protein kinase CK2alpha protein in 50 cases of LSCC tissues and 10 cases of normal mucous membrane of palatoglossal pillar were studied by using immunohistochemical SP staining method and quantitative analysis of image.. The positive rate of Protein kinase CK2alpha staining was 64% (32/50) in LSCC tissues,while only 30% (3/10) in normal mucous membrane of palatoglossal pillar (P <0.05). A significant difference was observed for the average absorbance value (A value) of protein kinase CK2alpha positive expression in two groups (P <0.05). No correlation was found between A value of protein kinase CK2alpha positive expression and sex, age, tumor sites, TNM stage and lymphatic metastasis, but there was significant correlation between A value of protein kinase CK2alpha positive expression and differentiation grading (G1-->G2 +G3) (P <0.05).. The over expression of Protein kinase CK2alpha is closely related to the formation and development of LSCC. Protein kinase CK2alpha may be one of the predictors for the malignant grade of LSCC. To inhibit the over expression might be new therapeutic methods for LSCC.

Topics: Aged; Carcinoma, Squamous Cell; Case-Control Studies; Casein Kinase II; Female; Humans; Laryngeal Neoplasms; Larynx; Lymphatic Metastasis; Male; Middle Aged; Neoplasm Staging

Gene expression profiling has been shown to be a valuable tool for prognostication and identification of cancer-associated genes in human malignancies. We aimed to identify potential prognostic marker(s) in non-small cell lung cancers using global gene expression profiles.. Twenty-one previously untreated patients with non-small cell lung cancer were analyzed using the Affymetrix GeneChip high-density oligonucleotide array and comparative genomic hybridization. Identified candidate genes were validated in an independent cohort of 45 patients using quantitative real-time reverse transcription-PCR and Western blot analyses. Follow-up data for these patients was collected and used to assess outcome correlations.. Hierarchical clustering analysis yielded three distinct subgroups based on gene expression profiling. Cluster I consisted of 4 patients with adenocarcinoma and 1 with squamous cell carcinoma (squamous cell carcinoma); clusters II and III consisted of 6 and 10 patients with squamous cell carcinoma, respectively. Outcome analysis was performed on the cluster groups containing solely squamous cell carcinoma, revealing significant differences in disease-specific survival rates. Moreover, patients having a combination of advanced Tumor-Node-Metastasis stage and assigned to the poor prognosis cluster group (cluster II) had significantly poorer outcomes. Comparative genomic hybridization analysis showed recurrent chromosomal losses at 1p, 3p, 17, 19, and 22 and gains/amplifications at 3q, 5p, and 8q, which did not vary significantly between the cluster groups. We internally and externally validated a subset of 11 cluster II (poor prognosis)-specific genes having corresponding chromosomal aberrations identified by comparative genomic hybridization as prognostic markers in an independent cohort of patients with lung squamous cell carcinoma identifying CSNK2A1 and C1-Inh as independent predictors of outcome.. CSNK2A1 and C1-Inh are independent predictors of survival in lung squamous cell carcinoma patients and may be useful as prognostic markers.

Topics: Aged; Biomarkers, Tumor; Blotting, Western; Carcinoma, Squamous Cell; Casein Kinase II; Chromosome Aberrations; Cluster Analysis; Cohort Studies; Complement C1 Inactivator Proteins; Complement C1 Inhibitor Protein; Cysteine Proteinase Inhibitors; Female; Follow-Up Studies; Gene Expression Profiling; Humans; Lung Neoplasms; Male; Nucleic Acid Hybridization; Oligonucleotide Array Sequence Analysis; Predictive Value of Tests; Prognosis; Protein Subunits; Survival Rate

Human squamous cell carcinomas of the head and neck (SCCHN) overexpress the protein kinase CK2, and elevated CK2 activity correlates with aggressive tumor behavior and poor clinical outcome. We therefore investigated whether interference with CK2 expression would inhibit SCCHN cell growth in vitro.. We targeted the catalytic (alpha) subunit of CK2 using an antisense oligodeoxynucleotide (ODN) strategy. Human Ca9-22 cells derived from SCCHN were transfected with CK2-alpha sense, nonsense, or antisense ODN; CK2 activity was measured; and the effect on CK2 activity and on cell growth was determined.. Transfection of Ca9-22 cells with antisense CK2-alpha ODN resulted in significantly decreased CK2 kinase activity associated with nuclear chromatin and in dose-dependent growth inhibition of Ca9-22 cells in vitro.. Interference with the protein kinase CK2 signal in SCCHN cells may offer a novel anticancer strategy for this malignancy.

Topics: Base Sequence; Carcinoma, Squamous Cell; Casein Kinase II; Cell Division; Codon, Nonsense; Dose-Response Relationship, Drug; Down-Regulation; Gene Expression; Gingival Neoplasms; Head and Neck Neoplasms; Humans; Oligonucleotides, Antisense; Probability; Protein Serine-Threonine Kinases; Reference Values; Sensitivity and Specificity; Transfection; Tumor Cells, Cultured

CK2 is a messenger-independent protein serine/threonine kinase that has been implicated in cell growth and proliferation. Our recent analysis of squamous cell carcinomas of the head and neck (SCCHN) revealed a significant elevation in CK2 activity in these tumor cells relative to normal mucosa of the upper aerodigestive tract and suggested a correlation with aggressive tumor behavior and poor clinical outcome. In order to further define the distribution of CK2 in these tissues, we have examined the immunohistochemical staining pattern of surgical specimens of both SCCHN tumors and normal upper aerodigestive tract mucosa using a monoclonal antibody directed against the catalytic subunit CK2-alpha of the kinase, and have compared these data with the subcellular distribution of CK2 activity in these same tissues. These measurements showed that CK2 is predominantly localized to the nuclei of the tumor cells, which agreed closely with the immunohistochemical staining pattern of CK2-alpha in tumor cells. The chiefly nuclear distribution of CK2-alpha immunostaining found consistently in SCCHN tumor cells and tumor-infiltrating lymphocytes contrasted with a relatively more predominant cytosolic staining pattern exhibited by various cellular constituents of normal oropharyngeal mucosa. The immunostaining pattern of CK2-alpha revealed that staining was observed in the cells stained for the proliferation-marker Ki-67; however, strong distinct immunostaining for CK2-alpha was also observed in large numbers of other cells in these same tumors, suggesting that CK2 elevation in these tumors is not a reflection of proliferative activity alone, but may also relate to the pathobiological behavior of the tumor.

Topics: Carcinoma, Squamous Cell; Casein Kinase II; Cell Division; DNA-Binding Proteins; Head and Neck Neoplasms; Humans; Immunohistochemistry; Protein Serine-Threonine Kinases; Tumor Cells, Cultured

We hypothesized that malignant transformation of normal mucosa of the upper aerodigestive tract to squamous cell carcinoma of the head and neck (SCCHN) might be associated with altered CK2 activity in the chromatin compartment of these tumors. We measured CK2 activity in the cytosol and chromatin of 7 surgical specimens of SCCHN, and 5 specimens of normal oropharyngeal mucosa from non-smokers/non-drinkers. CK2 activity in SCCHN tumors was significantly elevated in both the nuclear chromatin (P < 0.0005) and cytosolic (P <0.04) compartments relative to normal mucosa. These data suggest that activation of dysregulation of the chromatin-associated CK2 signal may play a role in the pathobiology od SCCHN.

Topics: Amino Acid Sequence; Base Sequence; Biomarkers, Tumor; Carcinoma, Squamous Cell; Casein Kinase II; Cell Nucleus; Cell Transformation, Neoplastic; Chromatin; Cytosol; Head and Neck Neoplasms; Humans; Molecular Sequence Data; Mouth Mucosa; Oligopeptides; Protein Serine-Threonine Kinases; Reference Values

Protein kinase CK2 (also known as casein kinase 2) is a messenger-independent protein serine/threonine kinase ubiquitously distributed in eukaryotes. CK2 has been found to phosphorylate a wide variety of cytosolic and nuclear substrates which are intimately involved in regulation of DNA, RNA, and protein synthesis, and differentiation. We therefore addressed the hypothesis that malignant transformation of upper aerodigestive tract mucosa to squamous cell carcinoma of the head and neck (SCCHN) might be associated with altered CK2 activity.. To this end, we subjected surgical specimens of SCCHN tumors and of normal oropharyngeal mucosa to subcellular fractionation. We then quantitated CK2 activity in cytosol and nuclei of these specimens using a CK2-specific peptide substrate (Arg-Arg-Arg-Glu-Glu-Glu-Thr-Glu-Glu-Glu).. We found that CK2 activity was significantly elevated in both nuclear (p < 0.0005) and cytosolic (p < 0.0034) compartments of SCCHN tumors, relative to normal oropharyngeal mucosa. Moreover, CK2 activity in the cellular cytosolic fraction of SCCHN tumors was associated with less differentiated histologic grade (p < 0.037), positive nodal metastatic status (p < 0.056), and a poor clinical outcome (p < 0.028). Kaplan-Meier cumulative survival analysis revealed greatly reduced survival in the high-CK2 activity patient group, with high statistical significance (p < 0.023).. These preliminary data reveal that malignant transformation of the upper aerodigestive tract mucosa is associated with altered CK2 activity. The results further suggest that dysregulation of this protein kinase may play a significant role in the pathobiology of SCCHN, and that CK2 activity may be a prognostic indicator in this malignancy.

Topics: Adult; Aged; Amino Acid Sequence; Biomarkers, Tumor; Carcinoma, Squamous Cell; Casein Kinase II; Cell Nucleus; Cell Transformation, Neoplastic; Cytosol; Female; Head and Neck Neoplasms; Humans; Lymphatic Metastasis; Male; Middle Aged; Molecular Sequence Data; Mucous Membrane; Oligopeptides; Oropharynx; Prognosis; Protein Serine-Threonine Kinases; Reference Values; Substrate Specificity; Survival Rate

Many of the effects of growth factors or hormones are mediated through the activation of protein kinase cascades. In this regard, it is well established that the activity of several protein kinases can be dramatically increased when cells are treated with a variety of stimuli. Since 1987, there have been several reports demonstrating that the activity of casein kinase II (CKII) can be acutely increased by hormones or growth factors. However, these are a number of discrepancies regarding the activation of CKII. In this study, we have examined CKII activities in extracts prepared from cells following treatment with stimuli that had been previously shown to elicit dramatic increases in CKII activity. Human WI.38 diploid lung fibroblasts were stimulated with serum or a variety of other stimuli including insulin, platelet-derived growth factor, fibroblast growth factor, epidermal growth factor, or phorbol myristate acetate. Human A431 epidermal carcinoma cells were similarly treated with epidermal growth factor. No reproducible increases in CKII activity were observed in response to any of these treatments. By comparison, a dramatic increase in kinase activity towards a synthetic peptide based on phosphorylation sites within the ribosomal S6 protein was consistently measured. Our observations indicate that CKII is not regulated in a similar manner by growth factors as are the protein kinases of the MAP kinase cascade, e.g., MAP kinase itself or ribosomal protein S6 kinase.

Topics: Amino Acid Sequence; Animals; Bombesin; Carcinoma, Squamous Cell; Casein Kinase II; Cattle; Cell Line; Cyclic AMP-Dependent Protein Kinases; Enzyme Activation; Enzyme Induction; Epidermal Growth Factor; Fetal Blood; Fibroblast Growth Factors; Fibroblasts; Growth Substances; Humans; Insulin; Ionomycin; Lung; Molecular Sequence Data; Neoplasm Proteins; Peptide Fragments; Phosphorylation; Platelet-Derived Growth Factor; Protein Processing, Post-Translational; Protein Serine-Threonine Kinases; Ribosomal Protein S6; Ribosomal Protein S6 Kinases; Ribosomal Proteins; Signal Transduction; Tetradecanoylphorbol Acetate; Tumor Cells, Cultured

In human epidermal carcinoma A431 cells, the beta subunit of casein kinase II is phosphorylated at an autophosphorylation site and at serine 209 which can be phosphorylated in vitro by p34cdc2 (Litchfield, D. W., Lozeman, F. J., Cicirelli, M. F., Harrylock, M., Ericsson, L. H., Piening, C. J., and Krebs, E. G. (1991) J. Biol. Chem. 266, 20380-20389). Given the importance of p34cdc2 in the regulation of cell cycle events, we were interested in examining the phosphorylation of casein kinase II during different stages of the cell cycle. In this study it is demonstrated that the extent of phosphorylation of serine 209 in the beta subunit is significantly increased relative to phosphorylation of the autophosphorylation site when chicken bursal lymphoma BK3A cells are arrested at mitosis by nocodazole treatment. This result suggests that serine 209 is a likely physiological target for p34cdc2. In addition, the alpha subunit of casein kinase II also undergoes dramatic phosphorylation with an associated alteration in its electrophoretic mobility when BK3A cells or human Jurkat cells are arrested with nocodazole. Phosphopeptide mapping studies indicate that p34cdc2 can phosphorylate in vitro the same peptides on the alpha subunit that are phosphorylated in cells arrested at mitosis. These phosphorylation sites were localized to serine and threonine residues in the carboxyl-terminal domain of alpha. Taken together, the results of this study indicate that casein kinase II is a probable physiological substrate for p34cdc2 and suggest that its functional properties could be affected in a cell cycle-dependent manner.

Topics: Animals; Antibodies; Carcinoma, Squamous Cell; Casein Kinase II; Cattle; CDC2 Protein Kinase; Cell Line; Chickens; Chromatography, Ion Exchange; Humans; Lymphoma; Macromolecular Substances; Male; Mitosis; Peptide Mapping; Phosphopeptides; Phosphorylation; Protein Serine-Threonine Kinases; Testis; Trypsin