Vinblastine and Griseofulvin Reversibly Disrupt the Living Mitotic Spindle

Malawista, Stephen E.; Sato, Hidemi; Bensch, Klaus G.

doi:10.1126/science.160.3829.770

Cited by 261 publications

(57 citation statements)

References 12 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This action of colchicine was considerably enhanced by cold which is known to cause the reversible breakdown of microtubules into subunits (30,31). In the present study we have found that two other drugs, demecolcine and vinblastine, which disrupt microtubules (32,33) also inhibit the release of histamine from leukocytes. At the same time D20 which is known to favor the formation of microtubules in other cells (8,9) markedly potentiates antigeninduced histamine release.…”

Section: Resultsmentioning

confidence: 75%

Histamine release from human leukocytes: studies with deuterium oxide, colchicine, and cytochalasin B

Gillespie¹,

Lichtenstein²

1972

J. Clin. Invest.

177

View full text Add to dashboard Cite

A B S T R A C T Agents known to interact with either microtubules or microfilaments influenced the antigen-induced release of histamine from the leukocytes of allergic individuals. Deuterium oxide (D20) which stabilizes microtubules and thereby favors their formation enhanced histamine release markedly. Concentrations as low as 5% increased antigen-induced release somnewhat while concentrations as high as 75% had no effect on release in the absence of antigen. Enhancement occurred over a wide range of antigen concentrations and was also seen when release was initiated by antibody to IgE or IgG. When the release process was divided into two stages a D20 activity could be demonstrated only in the second stage. However, when D20 was present in the first stage together with agents which raise cyclic AMP levels and thereby inhibit release it partially reversed this inhibition. Colchicine, demecolcine, and vinblastine, compounds known to disaggregate microtubules, i.e., have an effect opposite to that of D20, inhibited the release of histamine and counteracted the effects of D20. The inhibitory action of colchicine was greater if cells were treated with colchicine before rather than after activation with antigen. Cytochalasin B, a compound which causes the disappearance of microfilaments, had variable effects on histamine release. The most frequently seen response was slight enhancement. Neither D20 nor cytochalasin B altered cyclic AMP levels in leukocytes. These observations support and strengthen the view that an intact and functioning microtubule system is directly important for the secretion of histamine from leukocytes and suggest that microfilaments might have multiple indirect effects.

show abstract

Section: Resultsmentioning

confidence: 75%

Histamine release from human leukocytes: studies with deuterium oxide, colchicine, and cytochalasin B

Gillespie¹,

Lichtenstein²

1972

J. Clin. Invest.

177

View full text Add to dashboard Cite

show abstract

“…cal (1,12,16,23), biochemical, and physiological changes (5,(17)(18)(19)24) seen apparently as a direct consequence of this affinity between tubulin and the c-mitotic agents such as colchicine and the vinca alkaloids, vinblastine and vincristine. At the morphological level, a prominent manifestation of this affinity between some of the c-mitotic drugs and tubulin is seen in the destruction of the mitotic spindle and accumulation of cells in mitosis.…”

Section: Discussionmentioning

confidence: 99%

Cytofluorometric studies on the action of podophyllotoxin and epipodophyllotoxins (VM-26, VP-16-213) on the cell cycle traverse of human lymphoblasts.

Krishan¹,

Paika

Frei

1975

The Journal of Cell Biology

125

View full text Add to dashboard Cite

Flow microfluorometric analysis of human lymphoid cells exposed in vitro to cytostatic concentrations of podophyllotoxin (0.01-5 #g/ml for 24 h) shows that a major part of this population (40-60%) has the DNA content of cells in the G~-M part of the cell cycle, and that approximately 60% of these cells are arrested in mitosis. Although a similar pattern of DNA distribution is seen in cultures exposed to cytostatic concentrations of VM-26 (0.01 ~tg/ml) and VP-16-213 (0.1 ug/ml), no mitotic cells are seen in these cultures. Exposure to higher concentrations of VM-26 (0.1 ~g/ml) and VP-16-213 (1.0 ~g/ml) inhibits cell cycle traverse, and after 24 h of exposure a major part of the population is arrested with the DNA content of cells in the S part of the cell cycle. Exposure to higher drug concentrations leads to a reduction in the number of cells with the late S-G2 DNA content.Whereas the cell cycle block induced by cytostatic concentrations of podophyllotoxin (0.01 t~g/ml) is readily reversible by reincubation of cells in drug-free medium, cells blocked by VM-26 and VP-16-213 are unable to resume cell-cycle traverse under similar conditions. Medicinal properties of root extracts fromPodophyllum sp. have been known for more than a century. However, interest in the possible pharmacological and cancer chemotherapeutic properties of the various podophyllin derivatives has been currently revived by observations that topical application of podophyllin will cure condyloma acuminatum (9). A number of active ingredients have been isolated from podophyllin and include podophyllotoxin and 4-demethyl podophyllotoxin (10). An excellent review of the biochemical, pharmacological, and clinical literature on podophyllin published up to the year 1954 has been compiled by Kelly and Hartwell (10).Some of the earlier cytological studies have shown the colchicine-like effect of podophyllotoxin on the mitotic and meiotic spindles of both plant and animal cells (22). In eggs, destruction of the mitotic and meiotic spindles by exposure to podophyllotoxin, picropodophyllotoxin, and podophyllic acid has been reported (3). A number of subsequent reports have confirmed these observations and also described podophyllotoxin-induced nuclear fragmentation and necrosis (10).

show abstract

“…No enhancement in binding is detected with the DEAE-filter disc method of labeled colchicine. Vinblastine is thought to stabilize the native configuration of the protein and preserve colchicine-binding activity without directly affecting the binding site (19)(20)(21)(22). When the decay of colchicine-binding activity was monitored at 370 by fluorescence, it was observed that the protein lost only 18'% of its initial binding in the presence of 0.2 mM vinblastine sulfate, whereas without vinblastine, 76% of binding activity was lost over the same 7-hr interval.…”

Section: Methodsmentioning

confidence: 99%

Promotion of Fluorescence upon Binding of Colchicine to Tubulin

Bhattacharyya

Wolff

1974

Proc. Natl. Acad. Sci. U.S.A.

223

166

View full text Add to dashboard Cite

Colchicine, which does not fluoresce in aqueous media and organic solvents, exhibits marked fluorescence on combination with brain tubulin, with a corrected excitation maximum at 362 nm, an emission maximum at 435 nm, and a quantum yield of about 0.03.From fluorescence measurements it was found that rat brain tubulin binds 0.83 moles of colchicine per dimer (molecular weight 110,000) with an association constant of 3.2 ,M-1 at pH 7.0 A large body of evidence supports the view that microtubules from various sources are largely, if not completely, composed of tubulin, a dimeric protein of molecular weight 110,000-120,000 composed of two similar, but not identical, monomers (1). One of the most critical characteristics of tubulin is its ability to bind colchicine, an agent that leads to disaggregation of the polymerized form of this protein. The binding reaction of colchicine to tubulin has generally been studied by the use of labeled colchicine and subsequent separation of bound from free ligand by Sephadex gel filtration or ion exchange separation on DEAE-cellulose impregnated filter disc (2). Arai and Okuyama (3) recently pointed out that colchicine becomes fluorescent when bound to tubulin. We report here that fluorescence measurements offer a convenient method for measuring the colchicine tubulin interaction, which is based on the fact that colchicine fluoresces only in the bound form and not in the free state so that separation is not required. Thus, kinetic and thermodynamic parameters are easily obtained under equilibrium conditions. METHODS AND MATERIALSTubulin was prepared from rat brains in 10 mM Na phosphate (pH 7.0), 10 mM MgCl2, and by the procedure of Weisenberg et al. (2) except that DEAE-cellulose was used in place of DEAE-Sephadex. The purified tubulin solution was rapidly frozen in small aliquots and stored at -20°. The protein used gave a single band in sodium dodecyl sulfate-polyacrylamide gels (4), and its amino-acid analysis agreed well with that of Weisenberg et al. (2). The concentration of protein was determined by the method of Lowry et al. (5) spectrofluorometer, which gives corrected spectra in quanta band width. The quantum yields of colchicine-tubulin complexes were calculated by comparing the absorbances, at the exciting wavelength and the area of the emission spectra, of the colchicine-tubulin complex with quinine sulfate in 1.0 M H2SO4 whose quantum yield was taken as 0.546 at 250 (8).For convenience, routine analyses of tubulin-colchicine fluorescence were done with the Perkin-Elmer fluorometer. Calculation of Binding Data. Binding constants were obtained from fluorescence data by standard Scatchard analysis: r = (F,/FO) (CO/PO) where r = moles of colchicine boutd per mole of tubulin, Fc is the fluorescence of a given solution of colchicine-tubulin complex, and Fo is the fluorescence of an equal concentration of colchicine in excess tubulin, such that all the colchicine is bound, CO = the total colchicine concentration, and PO = the total protein concentration. From the ...

show abstract

Vinblastine and Griseofulvin Reversibly Disrupt the Living Mitotic Spindle

Cited by 261 publications

References 12 publications

Histamine release from human leukocytes: studies with deuterium oxide, colchicine, and cytochalasin B

Histamine release from human leukocytes: studies with deuterium oxide, colchicine, and cytochalasin B

Cytofluorometric studies on the action of podophyllotoxin and epipodophyllotoxins (VM-26, VP-16-213) on the cell cycle traverse of human lymphoblasts.

Promotion of Fluorescence upon Binding of Colchicine to Tubulin

Contact Info

Product

Resources

About