Reference - Detail
|Author||Bai T, Tanaka T, Yukawa K.|
|Title||Targeted knockdown of death-associated protein kinase expression induces TRAIL-mediated apoptosis in human endometrial adenocarcinoma cells.|
|Journal||Int J Oncol|
Death-associated protein kinase (DAPK) is a serine/threonine kinase that participates in the modulation of apoptosis and tumor suppression. Our previous study revealed high levels of DAPK protein expression in differentiated endometrial adenocarcinoma cells. To clarify the role of DAPK in human endometrial adenocarcinomas, we down-regulated endogenous DAPK expression in HHUA cells, a well-differentiated endometrial adenocarcinoma cell line, using specific small-interfering RNAs (siRNAs). The suppression of endogenous DAPK expression triggered apoptosis in HHUA cells, as evidenced by an increase in the sub-G1 DNA content in flow cytometric analyses. The apoptosis induced by the DAPK siRNA transfections was caspase-dependent, as characterized by the activations of caspase-3, -8 and -9. RNase protection assays detected higher levels of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), DR4 and DR5 transcripts in the DAPK siRNA-transfected HHUA cells than in the control siRNA-transfected cells. Consistent with these findings, enzyme-linked immunosorbent assays revealed that the DAPK siRNA transfections significantly increased the secretion of TRAIL protein from the cells. Treatment with recombinant human TRAIL protein dose-dependently suppressed the cell viability of HHUA cells. The present findings reveal that down-regulation of endogenous DAPK expression in HHUA cells induces caspase-dependent apoptosis, possibly through increased TRAIL, DR4 and DR5 signaling, thereby suggesting that DAPK expression is essential for HHUA cell survival. Consequently, endogenous DAPK mRNA may represent a potential candidate for molecularly targeted anticancer therapies.
|MeSH||Antineoplastic Agents / administration & dosage Antineoplastic Agents / pharmacology Apoptosis* / drug effects Apoptosis* / genetics Apoptosis Regulatory Proteins / antagonists & inhibitors Apoptosis Regulatory Proteins / genetics* Apoptosis Regulatory Proteins / physiology Calcium-Calmodulin-Dependent Protein Kinases / antagonists & inhibitors Calcium-Calmodulin-Dependent Protein Kinases / genetics* Calcium-Calmodulin-Dependent Protein Kinases / physiology Carcinoma, Endometrioid / drug therapy Carcinoma, Endometrioid / genetics Carcinoma, Endometrioid / pathology* Death-Associated Protein Kinases Drug Evaluation, Preclinical Drug Synergism Endometrial Neoplasms / drug therapy Endometrial Neoplasms / genetics Endometrial Neoplasms / pathology* Female Gene Expression Regulation, Enzymologic / drug effects Gene Expression Regulation, Enzymologic / physiology Gene Expression Regulation, Neoplastic / drug effects Gene Expression Regulation, Neoplastic / physiology Gene Knockdown Techniques Gene Targeting Humans RNA, Small Interfering / administration & dosage RNA, Small Interfering / pharmacology TNF-Related Apoptosis-Inducing Ligand / administration & dosage TNF-Related Apoptosis-Inducing Ligand / pharmacology* Transfection Tumor Cells, Cultured|
|Human and Animal Cells||HHUA(RCB0658)|