tae226 and Carcinoma--Squamous-Cell

To study the effect of the focal adhesion kinase inhibitor TAE226 on epithelial-mesenchymal transition (EMT) in human oral squamous cell carcinoma (OSCC) cell line.. HSC-3 and HSC-4 cells were cultured with TAE226 under different concentrations (0, 1, 5, and 10 μmol·L⁻¹) for 24, 48, and 72 h. Real-time quantitative polymerase chain reaction was performed to detect the mRNA expressions of E-cadherin and Vimentin. The protein expressions of E-cadherin and Vimentin were determined by Western blot assay after 48 h of TAE226 treatment.. Real-time quantitative polymerase chain reaction showed that increasing the TAE226 dose and reaction time resulted in increased and decreased E-cadherin and Vimentin mRNA expressions, respectively (P<0.05). Western blot assays showed that increasing the TAE226 dose resulted in increased and decreased E-cadherin and Vimentin protein expressions, respectively (P<0.05).. TAE226, which is expected to be an effective drug for OSCC treatment, can effectively inhibit the EMT of the OSCC cell line.. 目的 探究黏着斑激酶抑制剂TAE226对人口腔鳞状细胞癌细胞上皮间质转化（EMT）过程的影响。方法 不同浓度（0、1、5、10 μmol·L⁻¹）的TAE226作用于人口腔鳞状细胞癌HSC-3和HSC-4细胞24、48、72 h后，实时定量聚合酶链反应检测EMT标志物E-钙黏蛋白（E-cadherin）、波形蛋白（Vimentin）的mRNA表达；蛋白质印迹法检测E-cadherin、Vimentin在TAE226作用48 h后的蛋白表达。结果 实时定量聚合酶链反应检测表明，随着TAE226作用时间和浓度的增加，E-cadherin mRNA的表达增加，Vimentin mRNA的表达降低（P<0.05）。蛋白质印迹法检测表明，随着TAE226浓度的增加，E-cadherin蛋白的表达增加，Vimentin蛋白的表达降低（P<0.05）。结论 TAE226能有效抑制人口腔鳞状细胞癌细胞株的EMT进程，有望成为治疗口腔鳞状细胞癌的有效药物之一。.

Topics: Cadherins; Carcinoma, Squamous Cell; Cell Line, Tumor; Epithelial-Mesenchymal Transition; Focal Adhesion Protein-Tyrosine Kinases; Humans; Morpholines; Mouth Neoplasms; Vimentin

Focal adhesion kinase (FAK) overexpression is frequently found in invasive and metastatic cancers, but its role in oral squamous cell carcinoma is not yet well understood. In order to seek therapies targeting oral squamous cell carcinoma, we developed the novel FAK Tyr(397) inhibitor TAE226 and investigated its anti-tumor effects and mechanisms.. Expression of phosphorylated FAK Tyr(397) was examined by immunohistochemical and immunoblot analysis. The effect of TAE226 on in vitro and in vivo studies were confirmed by proliferation, cell cycle, apoptosis and angiogenesis analysis.. We found that phosphorylated FAK was highly expressed in human tongue oral squamous cell carcinoma in patients. Importantly, TAE226 greatly suppressed the proliferation, migration and invasion of human oral squamous cell carcinoma SAS cells with an apparent structural change of actin fiber and a loss of cell adhesion. In addition, TAE226 inhibited the expression of phospho-FAK Tyr(397) and phospho AKT Ser(473), resulting in caspase-mediated apoptosis. Furthermore, oral administration of TAE226 in mice suppressed the growth and angiogenesis of oral squamous cell carcinoma xenografts in vivo.. Our results provide compelling evidence that FAK is critically involved in oral squamous cell carcinoma and that the FAK inhibitor TAE226 can potentially be effectively used for the treatment of oral squamous cell carcinoma.

Topics: Animals; Apoptosis; Carcinoma, Squamous Cell; Cell Cycle; Cell Proliferation; Cell Transformation, Neoplastic; Female; Focal Adhesion Protein-Tyrosine Kinases; Humans; Mice; Mice, Inbred BALB C; Mice, Nude; Morpholines; Mouth Neoplasms; Phosphorylation; Proto-Oncogene Proteins c-akt

Epidermal growth factor receptor (EGFR) and focal adhesion kinase (FAK) show frequent overexpression and hyperactivity in various human malignancies including head and neck squamous cell carcinomas (HNSCC). To examine effects of dual EGFR/FAK inhibition on cellular radiosensitivity of HNSCC cells in a more physiological environment, we employed a previously established laminin-rich extracellular matrix (lrECM) based three-dimensional (3D) cell culture model.. UTSCC15 and SAS HNSCC cell lines stably transfected with EGFR-CFP or CFP were used. Single or combined EGFR (Cetuximab, siRNA) and FAK (TAE226, siRNA) inhibition were accomplished prior to measuring clonogenic survival and protein expression and phosphorylation. Immunofluorescence enabled visualization of EGFR-CFP and FAK.. Cetuximab resulted in higher radiosensitization in EGFR-CFP overexpressing cell lines than CFP controls. Single EGFR or FAK inhibition mediated radiosensitization, while dual EGFR/FAK targeting further augmented this effect. Despite signaling alterations upon Cetuximab and siRNA knockdown, analysis of protein expression and phosphorylation indicates EGFR and FAK signaling coexistence without obvious overlap.. Combined EGFR/FAK targeting yielded stronger radiosensitization than either approach alone, which might be based on non-overlapping downstream signaling. Whether dual targeting of EGFR and FAK can reasonably be combined with radiotherapy and chemotherapy needs clarification.

Topics: Antibodies, Monoclonal; Antibodies, Monoclonal, Humanized; Antineoplastic Agents; Apoptosis; Blotting, Western; Carcinoma, Squamous Cell; Cell Culture Techniques; Cell Line, Tumor; Cell Movement; Cell Proliferation; Cell Survival; Cetuximab; Dose-Response Relationship, Drug; ErbB Receptors; Focal Adhesion Protein-Tyrosine Kinases; Head and Neck Neoplasms; Humans; Morpholines; Phosphorylation; Radiation-Sensitizing Agents; Signal Transduction; Squamous Cell Carcinoma of Head and Neck; Transfection; Tumor Cells, Cultured

Focal adhesion kinase (FAK), a main player in integrin signaling and survival, is frequently overexpressed in human cancers and therefore postulated as potential target in cancer therapy. The aim of this study was to evaluate the radiosensitizing potential of the FAK inhibitor TAE226 in three-dimensional (3D) tumor cell cultures.. Head and neck squamous cell carcinoma (HNSCC) cells (FaDu, UT-SCC15, UT-SCC45), lung cancer cells (A549), colorectal carcinoma cells (DLD-1, HCT-116) and pancreatic tumor cells (MiaPaCa2, Panc1) were treated with different concentrations of TAE226 (0-1mum; 1 or 24h) without or in combination with irradiation (0-6Gy, X-ray, single dose). Subsequently, 3D clonogenic survival assays (laminin-rich extracellular matrix) and Western blotting (expression/phosphorylation, e.g. FAK, Akt, ERK1/2) were performed.. All investigated 3D cell cultures showed a dose-dependent reduction in clonogenic survival by TAE226. Intriguingly, TAE226 only significantly radiosensitized 3D HNSCC cell cultures accompanied by a pronounced dephosphorylation of FAK, Akt and ERK1/2.. Our data demonstrate TAE226 as potent FAK inhibitor that enhances the cellular radiosensitivity particularly of HNSCC cells grown in a 3D cell culture model. Future in vitro and in vivo investigations will clarify, to which extent this approach might be clinically relevant for radiotherapy of HNSCC.

Topics: Apoptosis; Carcinoma, Squamous Cell; Cell Movement; Cell Proliferation; Cell Survival; Colorectal Neoplasms; Dose-Response Relationship, Drug; Focal Adhesion Protein-Tyrosine Kinases; Head and Neck Neoplasms; Humans; Lung Neoplasms; Morpholines; Pancreatic Neoplasms; Probability; Radiation Tolerance; Radiation, Ionizing; Tumor Cells, Cultured