casein-kinase-ii and Laryngeal-Neoplasms

Laryngeal carcinoma is a common malignant neoplasm occurring in the head and neck, threatening human health. Protein casein kinase 2-alpha (CK2α) has been indicated to participate in the pathogenesis of this cancer; however, the underlying mechanisms still need to be elucidated. In this study, short hairpin RNA (shRNA)-mediated RNA interference (RNAi) technology was utilized to inhibit the CK2α expression in HEp-2 laryngeal carcinoma cells. Results showed that both mRNA and protein expression levels of endogenous CK2α were markedly decreased in HEp-2 cells transfected with CK2α specific shRNA. Transwell assays revealed that stable knockdown of CK2α significantly inhibited the migration and invasion of HEp-2 cells. As compared with cells treated with negative control shRNA, epithelial cadherin (E-cadherin) expression was increased, but snail, slug and vimentin were decreased in cells transfected with CK2α shRNA, indicating that inhibition of CK2α expression may suppress the epithelial-mesenchymal transition (EMT) process of laryngeal carcinoma in vitro. Moreover, suppression of CK2α was found to enhance the apoptosis induced by cisplatin in laryngeal carcinoma cells, probably through inhibition of permeability glycoprotein (P-glycoprotein) and multidrug-resistance protein (MRP1). In conclusion, our study may provide a promising therapeutic strategy for human laryngeal carcinoma by targeting CK2α.

Topics: Antigens, CD; Antineoplastic Agents; Apoptosis; Cadherins; Casein Kinase II; Cell Line, Tumor; Cell Movement; Cisplatin; Drug Screening Assays, Antitumor; Gene Expression; Gene Knockdown Techniques; Humans; Laryngeal Neoplasms; Neoplasm Invasiveness; RNA Interference; RNA, Small Interfering; Vimentin

The relationship between apoptosis and tumors is a major focus in cancer research. RNA interference (RNAi) technology has emerged as a very potent tool to generate cellular knockdown phenotypes of a desired gene. The aim of this study was to explore the effect of siRNA specific to the protein casein kinase 2α (CK2α) on apoptosis of laryngeal carcinoma cells and to explore possible mechanisms.. An siRNA expression plasmid specific to CK2α, psiRNA-hH1neo-CK2α, and a non-specific siRNA expression plasmid, psiRNA-hH1neo-cont, were constructed and transfected into Hep-2 cells by a lipofectamine method. The mRNA and protein levels of CK2α in transfected cells were determined by reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting analysis. Apoptosis was measured by Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) double-staining methods. The morphological changes to Hep-2 cells were observed under transmission electron microscopy (TEM). The levels of Bcl-2 and Bax proteins were measured by Western blotting analysis.. Levels of CK2α mRNA and protein were significantly decreased in the psiRNA-hH1neo-CK2α group compared to the other groups (P < 0.05). The apoptotic rate of the psiRNA-hH1neo-CK2α transfected group was significantly higher compared to that in the untransfected group and the siRNA-hH1neo-cont transfected group (25.66% ± 0.83%, 3.66% ± 0.43%, and 5.18% ± 0.22%) (P < 0.05). Compared with the untransfected group and the siRNA-hH1neo-cont transfected group, the psiRNA-hH1neo-CK2α transfected group presented with classical ultrastructural features of apoptosis, such as karyopyknosis, chromatic agglutination adjacent to the nuclear membrane, and apoptotic bodies. Compared with the other two groups, the level of Bcl-2 protein in the psiRNA-hH1neo-CK2α transfected group was decreased (0.20 ± 0.09 vs. 0.72 ± 0.16, 0.56 ± 0.11, P < 0.01), while the Bax protein level was increased (0.81 ± 0.17 vs. 0.26 ± 0.12, 0.33 ± 0.17, P < 0.01) and the ratio of Bcl-2 to Bax was decreased (0.25 ± 0.05 vs. 2.76 ± 0.21, 1.70 ± 0.22, P < 0.01).. The siRNA expression plasmid specific to CK2α could suppress CK2α expression and induce the apoptosis of laryngeal carcinoma cells. This is possibly by decreasing the Bcl-2/Bax ratio. CK2α may provide a potential therapeutic target against human laryngeal carcinoma.

Topics: Apoptosis; bcl-2-Associated X Protein; Blotting, Western; Casein Kinase II; Hep G2 Cells; Humans; Laryngeal Neoplasms; Microscopy, Electron, Transmission; Proto-Oncogene Proteins c-bcl-2; Reverse Transcriptase Polymerase Chain Reaction; RNA, Small Interfering

To investigate the effect of protein kinase CK2α on apoptosis and ultrastructure of human laryngeal carcinoma cells and its possible mechanism.. The siRNA expression plasmid psiRNA-hH1neo-CK2α specific to protein kinase CK2α and non-specific siRNA expression plasmid psiRNA-hH1neo-cont were transfected into Hep-2 cells respectively by lipofectamine method. Western blot was used to detect the expression of kinase CK2α protein. The apoptotic rate was measured by Annexin V-FITC/PI double-staining. The morphological changes of Hep-2 cells were observed under transmission electron microscope (TEM). The expressions of bcl-2 and Bax protein were measured by Western blot.. The expression of protein kinase CK2α protein significantly decreased in the Hep-2 cells transfected with psiRNA-hH1neo-CK2α (P < 0.01). Compared with the untransfected cells and siRNA-hH1neo-cont transfected group, psiRNA-hH1neo-CK2α transfected group presented with classical ultrastructural features of apoptosis, such as karyopyknosis, chromatic agglutination adjacent to nuclear membrane and apoptotic body. The apoptotic rate of psiRNA-hH1neo-CK2α transfected group was obviously higher than that in untransfected cells and siRNA-hH1neo-cont transfected group (25.66% ± 0.83% vs 3.66% ± 0.43%, 5.18% ± 0.22%, both P < 0.05). Compared with two other groups, the bcl-2 protein expression of psiRNA-hH1neo-CK2α transfected group decreased (0.20 ± 0.09 vs 0.72 ± 0.16, 0.56 ± 0.11, both P < 0.01), the Bax protein expression increased (0.81 ± 0.17 vs 0.26 ± 0.12, 0.33 ± 0.17, both P < 0.01) while the ratio of bcl-2 to Bax decreased (0.25 ± 0.05 vs 2.76 ± 0.21, 1.70 ± 0.22, both P < 0.01).. Protein kinase CK2a plays an important role in the apoptosis of human laryngeal carcinoma cells possibly by decreasing bcl-2/Bax. Protein kinase CK2a may provide a potential therapeutic target against human laryngeal carcinoma.

Topics: Apoptosis; Casein Kinase II; Cell Line, Tumor; Humans; Laryngeal Neoplasms; RNA, Small Interfering; Transfection

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

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

To assess the effect of small interfering RNA (siRNA) specific to protein kinase CK2a on proliferation and apoptosis of Hep-2 cell line.. siRNA expression plasmid psiRNA-hH1neo-CK2 specific to protein kinase CK2a and non-specific siRNA expression plasmid psiRNA-hH1neo-cont were constructed respectively, and then were transfected into Hep-2 cells by lipofectamine methods. Protein kinase CK2a mRNA and protein of the transfected cells were detected by reverse transcription polymerase chain reaction (RT-PCR) and Western Blot, respectively. Proliferation and apoptosis of the transfected cells were observed by methyl thiazolyl tetrazolium (MTT) method and flow cytometry (FCM), respectively.. Protein kinase CK2a mRNA and protein expressions were significantly decreased in the cells transfected with psiRNA-hH1neo-CK2 (P < 0.05). The Hep-2 cells grew slowly after transfected with psiRNA-hH1neo-CK2(P < 0.05). Obvious subdiploid peaks were found in the cells transfected with psiRNA-hH1neo-CK2 (P < 0.05).. siRNA expression plasmid specific to protein kinase CK2a suppressed the protein kinase CK2a expression and the proliferation of Hep-2, and induced apoptosis of Hep-2 cells.

Topics: Carcinoma; Casein Kinase II; Cell Line, Tumor; Cell Proliferation; Humans; Laryngeal Neoplasms; Plasmids; RNA, Messenger; RNA, Small Interfering; Transfection