ABSTRACT
Decursin is a major biological active component of Angelicagigas Nakai and is known to induce apoptosis of metastatic prostatic cancer cells. However, the apoptotic mechanism of decursin using primary malignant tumor (RC-58T/h/SA#4)-derived human prostate cells is not known. In the present study, we show that treatment of prostate cancer cells with decursin inhibited cell proliferation in a dose-dependent manner. Decursin also induced apoptosis in RC-58T/h/SA#4 cells, as determined by flow cytometry, Hoechst 33258 staining, and DNA fragmentation. Decursin caused activation of caspases-8, -9, and -3 and promoted the apoptotic action of caspase-8-mediated Bid cleavage. Decursin increased the protein levels of Bax and cytosolic cytochrome c as well as cleavage of PARP while decreasing the protein levels of Bcl-2. Furthermore, the caspase-independent mitochondrial apoptosis factor, apoptosis-inducing factor (AIF), was upregulated by treatment with decursin. Taken together, these findings indicate that decursin inhibited the proliferation of RC-58T/h/SA#4 cells through induction of apoptosis, which is mediated by both caspase-dependent and -independent apoptotic pathways.
Subject(s)
Angelica/chemistry , Apoptosis/drug effects , Benzopyrans/pharmacology , Butyrates/pharmacology , Caspases/metabolism , Prostatic Neoplasms/drug therapy , Bisbenzimidazole/chemistry , Cell Line, Tumor , Cell Survival/drug effects , DNA Fragmentation , Enzyme Activation/drug effects , Humans , Male , Microscopy, Fluorescence , Mitochondria/drug effects , Mitochondria/enzymology , Neoplasms, Hormone-Dependent/drug therapy , Neoplasms, Hormone-Dependent/metabolism , Neoplasms, Hormone-Dependent/pathology , Plant Roots/chemistry , Prostatic Neoplasms/metabolism , Prostatic Neoplasms/pathologyABSTRACT
Oat beta-glucosidase in plastid exists as a long fibrillar structure of AsGlu1 homomultimer (type I) and heteromultimer of AsGlu1 and AsGlu2 (type II). In spite of the high amino acid sequence homology of AsGlu1 and AsGlu2, AsGlu1 assembles into the fibrillar multimers but AsGlu2 forms a dimer when expressed in E. coli. A swapping analysis of AsGlu2 cDNA with AsGlu1 cDNA indicated that the C-terminal segment of AsGlu1 was critical for the fibrillar multimerization. A single substitution of glutamic acid-495 of AsGlu2 in the C-terminal region with lysine, an AsGlu1 counterpart amino acid for the glutamic acid-495, assembled the AsGlu2 into fibrillar homomultimers. The mutant AsGlu2 homomultimer was highly stable and had relatively faster electric mobility in native gel than the AsGlu1 homomultimer. Multimerization increased enzyme affinity to substrates.
