Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:3.4.22.62 (caspase-9)
 7,507 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

RNA interference is an evolutionarily conserved cellular defense mechanism that protects cells from hostile genes and regulates the function of normal genes during growth and development. In this study, we established GFP-siFAK-DLC1 vector and transfect the vector into OVCAR-3 cells. RT-PCR and western blot analyses were performed for FAK, DLC1 mRNA, and protein expression in OVCAR-3 cells. ELISA method was used for caspase-3 and caspase-9 activities. These studies demonstrate that both recombinant pGFP-siFAK-DLC1 vector and pGFP-siCon-DLC1 vector may effectively promote DLC1 mRNA transcription and didn't affect siRNA effect. Recombinant vector (pGFP-siFAK-DLC1) may promote DLC1 gene expression, and effectively silence FAK gene expression. Silencing FAK mRNA expression and DLC1 mRNA expression may markedly enhance caspase-3 and caspase-9 activities in OVCAR-3 cells. These results showed that in ovarian cancer OVCAR-3 cell silencing FAK gene expression or / and increasing DLC-1 gene expression, could improve Caspase-3 and Caspase-9 protease activities. In the expression of DLC-1 and silence FAK expression group (double action group) effect was more significant as compared with the silence FAK gene group or expression of DLC-1 gene alone, difference was significant (p < 0.05).
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PMID:Effect of small interfering RNA transfection on FAK and DLC1 mRNA expression in OVCAR-3. 2276 Feb 57

Previous studies proposed that myosin-Va regulates apoptosis by sequestering pro-apoptotic Bmf to the actin cytoskeleton through dynein light chain-2 (DLC2). Adhesion loss or other cytoskeletal perturbations would unleash Bmf, allowing it to bind and inhibit pro-survival Bcl2 proteins. Here, we demonstrated that overexpression of a myosin-Va medial tail fragment (MVaf) harboring the binding site for DLC2 dramatically decreased melanoma cell viability. Morphological and molecular changes, including surface blebbing, mitochondrial outer membrane permeabilization, cytochrome-c and Smac release, as well as caspase-9/-3 activation and DNA fragmentation indicated that melanoma cells died of apoptosis. Immobilized MVaf interacted directly with DLCs, but complexed MVaf/DLCs did not interact with Bmf. Overexpression of DLC2 attenuated MVaf-induced apoptosis. Thus, we suggest that, MVaf induces apoptosis by sequestering DLC2 and DLC1, thereby unleashing the pair of sensitizer and activator BH3-only proteins Bmf and Bim. Murine embryonic fibroblasts (MEFs) lacking Bim and Bmf or Bax and Bak were less sensitive to apoptosis caused by MVaf expression than wild-type MEFs, strengthening the putative role of the intrinsic apoptotic pathway in this response. Finally, MVaf expression attenuated B16-F10 solid tumor growth in mice, suggesting that this peptide may be useful as an apoptosis-inducing tool for basic and translational studies.
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PMID:A myosin-Va tail fragment sequesters dynein light chains leading to apoptosis in melanoma cells. 2351 16