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      Rotenoids mediate potent cancer chemopreventive activity through transcriptional regulation of ornithine decarboxylase.

      Nature medicine

      antagonists & inhibitors, 9,10-Dimethyl-1,2-benzanthracene, Animals, Antineoplastic Agents, Phytogenic, pharmacology, Cell Differentiation, drug effects, Cells, Cultured, Female, Gene Expression Regulation, Enzymologic, HL-60 Cells, cytology, Humans, Mice, Mice, Inbred BALB C, Neoplasms, Experimental, enzymology, pathology, prevention & control, Organ Culture Techniques, Ornithine Decarboxylase, genetics, Precancerous Conditions, Protein Kinase C, metabolism, RNA, Messenger, Rotenone, analogs & derivatives, Skin Neoplasms, chemically induced, Tetradecanoylphorbol Acetate

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          Abstract

          For the discovery of new cancer chemopreventive agents, we have studied the potential of plant extracts to inhibit phorbol ester-induced ornithine decarboxylase (ODC) activity in cell culture. Four active rotenoids were obtained from the African plant Mundulea sericea (Leguminosae). These isolates were highly potent when evaluated for inhibition of chemically induced preneoplastic lesions in mammary organ culture and inhibition of papillomas in the two-stage mouse skin model, and they appear to function by a unique mechanism at the level of ODC messenger RNA expression. Based on our findings, rotenoids can be regarded as promising new chemopreventive or anticancer agents.

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          Most cited references 8

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          Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction.

          A new method of total RNA isolation by a single extraction with an acid guanidinium thiocyanate-phenol-chloroform mixture is described. The method provides a pure preparation of undegraded RNA in high yield and can be completed within 4 h. It is particularly useful for processing large numbers of samples and for isolation of RNA from minute quantities of cells or tissue samples.
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            Ornithine decarboxylase activity is critical for cell transformation.

            The enzyme ornithine decarboxylase is the key regulator of the synthesis of polyamines which are essential for cell proliferation. Expression of this enzyme is transiently increased upon stimulation by growth factors, but becomes constitutively activated during cell transformation induced by carcinogens, viruses or oncogenes. To test whether ornithine decarboxylase could be a common mediator of transformation and oncogenic itself, we transfected NIH3T3 cells with expression vectors carrying the complementary DNA encoding human ornithine decarboxylase in sense and antisense orientations. The increased expression of the enzyme (50-100-times endogenous levels) induced not only cell transformation, but also anchorage-independent growth in soft agar and increased tyrosine phosphorylation of a protein of M(r) 130K. Expression of ornithine decarboxylase antisense RNA was associated with an epithelioid morphology and reduced cell proliferation. Moreover, blocking the endogenous enzyme using specific inhibitor or synthesizing antisense RNA prevented transformation of rat fibroblasts by temperature-sensitive v-src oncogene. Our results imply that the gene encoding ornithine decarboxylase is a proto-oncogene central for regulation of cell growth and transformation.
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              • Record: found
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              Rotenone inhibition of tubulin self-assembly.

              Rotenone effectively inhibits the in vitro formation of microtubules from tubulin containing or lacking microtubule-associated proteins. In both cases a concentration of rotenone equal to that of tubulin present completely blocks assembly. The inhibition can be reversed by the addition of dimethylsulfoxide or by removing rotenone with charcoal.
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                7585044

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