1. Induction of apoptosis against cancer cell lines by four ascomycetes (endophytes) from Malaysian rainforest
Kalavathy Ramasamy, Anthony L J Cole, Siong Meng Lim, Abu Bakar Abdul Majeed, Nurul Aqmar Mohamad Nor Hazalin Phytomedicine . 2012 May 15;19(7):609-17. doi: 10.1016/j.phymed.2012.01.007.
Endophytic fungi have been shown to be a promising source of biologically active natural products. In the present study, extracts of four endophytic fungi isolated from plants of the National Park, Pahang were evaluated for their cytotoxic activity and the nature of their active compounds determined. Those extracts exhibiting activity with IC(50) values less than 17 μg/ml against HCT116, MCF-7 and K562 cell lines were shown to induce apoptosis in these cell lines. Molecular analysis, based on sequences of the rDNA internal transcribed spacers ITS1 and ITS4, revealed all four endophytic fungi to be ascomycetes: three sordariomycetes and a dothideomycete. Six known compounds, cytochalasin J, dechlorogriseofulvin, demethylharzianic-acid, griseofulvin, harzianic acid and 2-hexylidene-3-methyl-succinic acid were identified from a rapid dereplication technique for fungal metabolites using an in-house UV library. The results from the present study suggest the potential of endophytic fungi as cytotoxic agents, and there is an indication that the isolates contain bioactive compounds that mainly kill cancer cells by apoptosis.
2. Redistribution of motor proteins by Cytochalasin J treatment
Judith A Snyder, Rollin W Robinson Cell Biol Int . 2003;27(8):665-73. doi: 10.1016/s1065-6995(03)00123-9.
In this study we extend our analysis of the effect of Cytochalasin J (CJ) on mitotic and interphase cells by the use of immunocytochemical techniques to localize antigens to anti-beta-tubulin, anti-dynein heavy chain (HC), anti-dynein intermediate chain (IC), and anti-kinesin antibodies following CJ treatment. Anti-dynein IC and HC staining of CJ treated cells showed a significant reduction in anti-dynein staining in the nuclear region of interphase cells. Monolayer cultures of PtK(1)cells treated with 10 microg/ml CJ for 10 min showed a significant reduction in pixel luminosity of fluorescence staining using anti-dynein IC and HC antibodies (P<0.05). Cytochalasin J treatment reorganized anti-dynein staining from a cytoplasmic punctate staining with greatest intensity in the perinuclear region, to a more uniform staining throughout the cytoplasm.
3. Cytochalasin J affects chromosome congression and spindle microtubule organization in PtK1 cells
L Cohen, J A Snyder Cell Motil Cytoskeleton . 1995;32(4):245-57. doi: 10.1002/cm.970320402.
PtK1 cells were treated with 10 micrograms/ml cytochalasin J (CJ) for 15 min at various stages of mitosis. When applied at nuclear envelope breakdown (NEB) chromosome congression was blocked or substantially slowed, and chromosomes failed to show organization patterns typical of prometaphase. Spindle microtubule (MT) numbers appeared unaffected as judged by the pattern of birefringent retardation. However, ultrastructural analysis showed MTs to be reorganized within the spindle domain with some exhibiting fragmentation and others failing to interact with poorly defined kinetochore laminae. The spindle domain took on a curved, almost banana-like shape, as related to the position of the centrosomes and lack of orientation of chromosomes. Serial section analysis of kinetochore regions showed reduced contour length and maturation of the kinetochore plate with few MTs associated with this structure. Cells similarly treated with 10 micrograms/ml CJ at NEB for 15 min and then released into conditioned medium for 15 min showed the most chromosomes resumed congression to the metaphase plate. Ultrastructural analysis revealed a more normal organization of spindle MTs, but kinetochore structure remained affected. CJ treatment of cells in prometaphase slightly affected chromosomes congression with most chromosomes aligning at the metaphase plate after 10-15 min of treatment. Ultrastructural analysis showed that astral MTs were disrupted and spindle MTs were fragmented; few MTs coursed from kinetochore to pole. Kinetochore structure was also affected with only small numbers of short MTs seen associated with kinetochores. Application of CJ at anaphase onset had little effect on anaphase A and B, but cytokinesis failed to occur. Anti-tubulin staining of a monolayer of cells treated with 10 micrograms/ml CJ for 15 min showed that over 60% of mitotic figures exhibited changes in MT organization. Cells showing the greatest effect of treatment had several foci of bundles of MTs, as if the spindle were multipolar. Chromosomes were arranged near the periphery of the spindle which could be a result of abnormalities of kinetochore structure. Improper association of spindle MTs with kinetochores and, thus, changes in kinetochore position could account for these changes in spindle architecture.
