Taxol transport in Taxus baccata cell suspension cultures

Fornalé, Silvia; Esposti, Davide Degli; Navia-Osorio, Alberto; Cusidó, Rosa M.; Palazón, Javier; Piñol, M. Teresa; Bagni, Nello

doi:10.1016/s0981-9428(01)01332-8

Cited by 28 publications

(19 citation statements)

References 33 publications

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“…These results suggest that the stationary phase was extremely short, as cell death rapidly followed exponential growth phase. These results do not agree with those reported by Fornalè et al (2002) for Taxus baccata, that suggest that despite the decrease in cell division during the stationary phase, cell viability should remain high. During the growth of P. pinaster suspension cell cultures, changes in medium pH were observed (Fig.…”

Section: Resultscontrasting

confidence: 87%

“…suspension cultures (Fornalè et al 2002). Sugar consumption in P. pinaster suspension cell cultures is depicted in Fig.…”

Section: Sugar Consumptionmentioning

confidence: 99%

“…Even though work with conifer species is scarce when compared to angiosperms, gymnosperm cell suspensions have been successfully used in many studies addressing the biochemistry and molecular biology of primary (Ishii and Teasdale 1997) and secondary metabolism (Fornalè et al 2002), biotic stress (Cooney et al 2000) and abiotic stress (Chiron et al 2000). More often, suspension cell cultures are the first step in the establishment of plant regeneration systems (Ingram and Mavituna 2000).…”

Section: Introductionmentioning

confidence: 99%

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Establishment and characterization of Pinus pinaster suspension cell cultures

Azevedo

Dias

Tavares

2008

Plant Cell Tiss Organ Cult

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We report the establishment of a Pinus pinaster (Ait.) cell suspension culture in a modified MS medium supplemented with 2 mg ml -1 2,4-D and 1 mg ml -1 BA. Calli were obtained from seedling root segments and established a friable isodiametric cell suspension, suitable for in vitro studies of maritime pine at the cellular level. Growth (dry weight), cell viability, pH, and nutrient consumption: carbon source (sucrose, fructose and glucose), nitrogen source (ammonia and nitrate) and phosphate were monitored over 24 h. Suspension cells exhibited a 15-day exponential growth stage, during which a biphasic consumption profile was observed for all nutrients. Phosphate was the first limiting nutrient and preferable consumption was observed for glucose over fructose and nitrate over ammonium.

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Section: Resultscontrasting

confidence: 87%

“…suspension cultures (Fornalè et al 2002). Sugar consumption in P. pinaster suspension cell cultures is depicted in Fig.…”

Section: Sugar Consumptionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Establishment and characterization of Pinus pinaster suspension cell cultures

Azevedo

Dias

Tavares

2008

Plant Cell Tiss Organ Cult

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“…39) In contrast, Fornalè reported that the exogenous addition of taxol results in its accumulation in the vacuoles rather than the cell wall of T. baccata cell suspension. (3) In addition, an export mechanism of taxol may be present in Taxus species, 40) and (4) a taxol may undergo a structural modification, which could reduce its toxicity and create conditions in which the microtubule is not affected by the toxin cocktail. All of these mechanisms would proceed in combination with each other and contribute to the selfresistance of T. cuspidata.…”

Section: Discussionmentioning

confidence: 99%

Expression of recombinant alpha and beta tubulins from the yew Taxus cuspidata and analysis of the microtubule assembly in the presence of taxol

Kudo

Abe

Ito

et al. 2014

Bioscience, Biotechnology, and Biochemistry

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Taxol was originally isolated from the yew Taxus brevifolia. Because taxol inhibits the depolymerization of microtubules, the presence of a self-resistance mechanism in Taxus spp. was hypothesized. The cloning of the cDNA for alpha and beta tubulins from Taxus cuspidata and those from the human embryonic kidney cell line HEK293T revealed that the 26Asp, 359Arg, and 361Leu residues in the human beta tubulin, which are important for taxol binding, were replaced with Glu, Trp, and Met in the beta tubulin of T. cuspidata, respectively. The microtubule assembly of the recombinant alpha and beta tubulins was monitored turbidimetrically, and the results clearly demonstrated that the microtubule from T. cuspidata is less sensitive to taxol than that from HEK293T cells. The Taxus microtubule composed of the wild-type alpha tubulin and the beta tubulin with the E26D mutation restored the sensitivity to taxol. We thus postulated that the mutation identified in the beta tubulin of T. cuspidata plays a role in the self-resistance of this species against taxol.

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“…In some cases, a targeted engineering approach aimed at a specific secondary metabolite transporter has proven to be successful [26], but this has been relatively rare due to the lack of knowledge regarding most transport systems as well as difficulty in engineering some plant systems, particularly medicinal plant species such as Taxus. There have been several studies aimed at paclitaxel transport in Taxus systems, including uptake studies utilizing [ 14 -C]-paclitaxel as a tracer, which suggested there was an active, energy-dependent paclitaxel uptake mechanism [27], as well as studies utilizing a spin probe which indicated a decreased membrane permeability in Taxus x media upon elicitation with jasmonic acid [28]. The identification of a specific transport mechanism for paclitaxel has yet to be demonstrated.…”

Section: Introductionmentioning

confidence: 99%

Paclitaxel uptake and transport in Taxus cell suspension cultures

Naill

Kolewe

Roberts

2012

Biochemical Engineering Journal

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The transport of paclitaxel in Taxus canadensis suspension cultures was studied with a fluorescence analogue of paclitaxel (Flutax-2®) in combination with flow cytometry detection. Experiments were carried out using both isolated protoplasts and aggregated suspension cell cultures. Flutax-2® was shown to be greater than 90% stable in Taxus suspension cultures over the required incubation time (24 hours). Unlabeled paclitaxel was shown to inhibit the cellular uptake of Flutax-2®, although structurally similar taxanes such as cephalomannine, baccatin III, and 10-deacetylbaccatin III did not inhibit Flutax-2® uptake. Saturation kinetics of Flutax-2® uptake was demonstrated. These results indicate the presence of a specific transport system for paclitaxel. Suspension cells elicited with methyl jasmonate accumulated 60% more Flutax-2® than unelicited cells, possibly due to an increased cellular storage capacity following methyl jasmonate elicitation. The presence of a specific mechanism for paclitaxel transport is an important first result that will provide the basis of more detailed studies as well as the development of targeted strategies for increased paclitaxel secretion to the extracellular medium.

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Taxol transport in Taxus baccata cell suspension cultures

Cited by 28 publications

References 33 publications

Establishment and characterization of Pinus pinaster suspension cell cultures

Establishment and characterization of Pinus pinaster suspension cell cultures

Expression of recombinant alpha and beta tubulins from the yew Taxus cuspidata and analysis of the microtubule assembly in the presence of taxol

Paclitaxel uptake and transport in Taxus cell suspension cultures

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