Conjugated linoleic acid inhibits Caco-2 cell growth via ERK-MAPK signaling pathway.

Conjugated linoleic acid (CLA) is a naturally occurring compound found in dairy and beef products. In recent years, it has received considerable attention because several studies showed a lower incidence of certain cancers in animals fed CLA-supplemented diets. In vitro studies further showed growth inhibitory activity on tumor cell proliferation, the CLA being effective above all against colon cancer cells. The aim of the present work was to investigate the growth inhibitory effect of CLA on Caco-2 cell line. Under our experimental conditions, CLA repressed Caco-2 cell proliferation, and the growth-inhibitory action increased by repeating treatments. However, in Caco-2 cells, CLA was unable to induce apoptosis, as revealed by cell-cycle analysis and Western blot studies. To determine the mechanism by which CLA inhibits cell growth, we studied its effect on extracellular-regulated kinase signaling. Conjugated linoleic acid reduced expression levels of Raf-1 and phosphorylation of ERK1/2, which was accompanied by a decrease in the expression of the downstream transcription factor c-myc. Our data suggest that CLA is dependent, at least in part, on the ERK kinase pathway for its ability to inhibit the growth of Caco-2 cancer cells.

Extracellular signal-regulated kinase 1/2 and protein phosphatase 2A are involved in the antiproliferative activity of conjugated linoleic acid in MCF-7 cells.

Conjugated linoleic acid (CLA) has protective properties in breast cancer. Here, we studied the mechanisms underlying the effects of CLA on MCF-7 breast cancer cell proliferation, especially in correlation with the involvement of the extracellular signal-regulated kinase 1/2 (ERK1/2) pathway and protein phosphatase 2A (PP2A). CLA inhibits MCF-7 cell growth in a concentration- and time-dependent manner, without triggering apoptosis. In assessing expression levels of proteins that play obligatory roles in the ERK cascade, we evidenced that CLA down-regulated Raf-1 and decreased levels of phospho-ERK1/2, as well as c-myc expression. Increase in PP2A expression rates were additionally observed after CLA treatment of MCF-7 cells. The above effects, as well as CLA-induced inhibition of cell growth, were reversed by okadaic acid, a specific inhibitor of PP2A. Thus, PP2A likely participates in deactivation of ERK1/2, and its up-regulation may represent a novel mechanism for CLA-induced inhibition of cell proliferation.

Conjugated linoleic acid induces apoptosis in MDA-MB-231 breast cancer cells through ERK/MAPK signalling and mitochondrial pathway.

We investigated the molecular mechanisms involved in the anti-proliferative activity exerted by conjugated linoleic acid (CLA) on the estrogen unresponsive MDA-MB-231 human breast cancer cell line. The effects on cell proliferation, cell cycle progression and induction of apoptosis were examined. CLA caused the reduction of cell proliferation along with the accumulation of cells in the S phase of the cycle. The occurrence of apoptosis in these cells was indicated by flow cytometry data and further confirmed by the onset of cells with morphological features typical of apoptosis. ERK1/2 reduction and upregulation of pro-apoptotic protein Bak were induced. These events were associated with: (a) reduced levels of the anti-apoptotic protein Bcl-x(L), (b) the translocation of cytochrome c from the mitochondria to the cytosol, (c) the cleavage of pro-caspase-9 and pro-caspase-3. From the above data, we are induced to think that CLA may trigger apoptosis in the estrogen unresponsive MDA-MB-231 cell line via mechanisms involving above all the mitochondrial pathway.

Microtubule-interfering activity of parthenolide.

Parthenolide is an active sesquiterpene lactone present in a variety of medicinal herbs, well known as anti-inflammatory drug. It has recently been proposed as a chemotherapeutic drug, but the pharmacological pathways of its action have not yet been fully elucidated. Firstly, we explored whether the anticancer properties of parthenolide may be related to a tubulin/microtubule-interfering activity. We additionally compared bioactivities of parthenolide with those checked after combined treatments with paclitaxel in human breast cancer MCF-7 cells. Parthenolide exerted in vitro stimulatory activity on tubulin assembly, by inducing the formation of well-organized microtubule polymers. Light microscopy detections showed that parthenolide-induced alterations of either microtubule network and nuclear morphology happened only after combined exposures to paclitaxel. In addition, the growth of MCF-7 cells was significantly inhibited by parthenolide, which enhanced paclitaxel effectiveness. In conclusion, the antimicrotubular and antiproliferative effects of parthenolide, well known microtubule-stabilizing anticancer agent, may influence paclitaxel activity. The tubulin/microtubule system may represent a novel molecular target for parthenolide, to be utilized in developing new combinational anticancer strategies.

Insulin can modulate MCF-7 cell response to paclitaxel.

Insulin regulates metabolism through homologous receptor tyrosine kinases, and plays a role in proliferation of breast cancer cells. Our research studied whether insulin, administered separately or in combination with paclitaxel, interferes with paclitaxel-mediated biological activity in human breast cancer cells. Not only did insulin influence paclitaxel-mediated cell microtubule reorganization, but it also influenced MCF-7 cell sensitivity to paclitaxel. Furthermore, combined administrations of insulin and paclitaxel affected MAPK pathway, Raf-1 activation and p53 expression levels. Our findings indicate that insulin seems to modulate MCF-7 cell response to paclitaxel; consequently, elevated levels of insulin could influence tumor cell resistance.

A sesquiterpene lactone, costunolide, interacts with microtubule protein and inhibits the growth of MCF-7 cells.

Costunolide is an active sesquiterpene lactone of medicinal herbs with anti-inflammatory and potential anti-cancer activity. Nevertheless, the pharmacological pathways of costunolide have not yet been fully elucidated. In this study we showed that costunolide exerts a dose-dependent antiproliferative activity in the human breast cancer MCF-7 cells. In addition, light microscopy observations indicated that costunolide affected nuclear organization and reorganized microtubule architecture. The antiproliferative and antimicrotubular effects of costunolide were not influenced by paclitaxel, well-known microtubule-stabilizing anticancer agent. The microtubule-interacting activity of costunolide was confirmed by in vitro studies on purified microtubular protein. In fact, costunolide demonstrated polymerizing ability, by inducing the formation of well organized microtubule polymers. Our data suggest an interaction of costunolide with microtubules, which may represent a new intracellular target for this drug.

Biological properties of jatrophane polyesters, new microtubule-interacting agents.

PURPOSE: The biological activities of macrocyclic jatrophane polyesters 1-3 from the Sardinian endemism Euphorbia semiperfoliata Viv. have not been evaluated in depth. We investigated the microtubule-interacting and antiproliferative activities of these drugs and the molecular mechanisms underlying their effects. METHODS: We tested jatrophanes for their interaction with purified bovine brain tubulin by an in vitro polymerization assay and by electron microscopy. At a cellular level, the effects of jatrophanes on microtubular architecture, nuclear morphology, cell viability, cell cycle perturbations, and p53 and Raf-1/Bcl-2 involvement were investigated. RESULTS: Jatrophanes exhibited microtubule-interacting activity. They stimulated purified tubulin assembly in vitro, and induced paclitaxel-like microtubules, as revealed by electron microscopy. In the cells, rearrangement of microtubule architecture was in contrast to the bundling produced by paclitaxel. Jatrophanes inhibited the growth of some human cancer cell lines without inducing cell cycle arrest in the G2/M phase. Moreover, they influenced p53 expression and Raf-1/Bcl-2 activation. CONCLUSIONS: Despite their structural difference from paclitaxel and other microtubule-interacting agents, jatrophanes may represent a new type of tubulin binder.

Microtubule-interacting activity and cytotoxicity of the prenylated coumarin ferulenol.

In this study we investigated biological activity of ferulenol, a prenylated 4-hydroxycoumarin from Ferula communis. Ferulenol stimulates tubulin polymerization in vitro, and inhibits the binding of radiolabeled colchicine to tubulin. It rearranges cellular microtubule network into short fibres, and alters nuclear morphology. Remarkably, ferulenol exerts a dose-dependent cytotoxic activity against various human tumor cell lines. Abbreviations. DAPI:4,6-diamidino-2-phenylindole DMEM:Dulbecco's modified Eagle's medium EGTA:ethylenebis(oxyethylenenitrilo)tetraacetic acid FBS:fetal bovine serum FITC:fluoresceine isothiocyanate GTP:guanosine 5'-triphosphate MES:2-[N-morpholino]ethane-sulfonic acid MTP:microtubule protein SDS:sodium dodecyl sulfate

Comparative studies on biological activity of certain microtubule-interacting taxanes.

We have compared the nature of interaction of certain taxanes with microtubular protein, and the mechanism of action underlying cytotoxic activity. Taxanes induced tubulin assembly in vitro, but only taxanes bearing side chain were capable of inducing the formation of stable tubulin polymers. Electron microscopy detections showed that taxane-induced polymers are structurally similar to microtubules formed by paclitaxel, with differences in length. Otherwise, light microscopy views have shown that intracellular microtubule network is deeply reorganized by taxanes into short fibers, unlike paclitaxel-bundled microtubules. Taxanes inhibited the growth of various human tumor cell lines, but cell cycle analysis did not always indicate a block in the G2/M phase. These agents alter some apoptotic signal transduction pathways, probably by a mechanism distinct from microtubule interaction. Briefly, the effectiveness of taxanes is closely related to their chemical structure, and depends on their interaction with microtubular protein. By virtue of this mechanism, some of these taxanes may provide usefulness for therapeutic improvements.