Inactivation of wild-type p53 by a dominant negative mutant renders MCF-7 cells resistant to tubulin-binding agent cytotoxicity

Galmarini, Carlos M.; Falette, Nicole; Tabone, Eric; Levrat, Christiane; Britten, Richard A.; Voorzanger-Rousselot, Nathalie; Roesch-Gateau, O; Vanier-Viornery, A; Puisieux, Alain; Dumontet, Charles

doi:10.1054/bjoc.2001.2017

Cited by 37 publications

(33 citation statements)

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“…We have previously shown that MDD2 cells displayed 31.5-fold resistance to paclitaxel (MN-1 IC 50 : 7.6 nM; MDD2 IC 50 : 240 nM) and 50-fold resistance to vinblastine (MN-1 IC 50 : 6.8 nM; MDD2 IC 50 : 340 nM) in comparison with MN-1 cells (Galmarini et al, 2001). …”

Section: Cell Linesmentioning

confidence: 97%

“…HCT116 p53À/À is a cell line derived from colorectal carcinoma in which two promoterless targeting vectors were used to disrupt sequentially the two p53 alleles (Bunz et al, 1998). All cell lines were cultured as previously described (Galmarini et al, 2001).…”

Section: Cell Linesmentioning

confidence: 99%

mentioning

confidence: 99%

“…In cancer cells, alterations of the p53 tumour suppressor gene confer resistance to microtubule-targeted drugs (Wu and El-Diery, 1996;Galmarini et al, 2001). We recently reported that chemoresistance present in p53-deficient cells might be attributed to defective apoptotic pathways (Galmarini et al, 2001). However, p53 also associates with microtubules in vitro and in vivo (Giannakakou et al, 2000b), and has been reported to regulate negatively the putative microtubule-stabilising protein microtubule-associated protein 4 (MAP4), the microtubule-related inhibitor of apoptosis survivin (Mirza et al, 2002), and the microtubule-destabilising protein stathmin/OP18 (Johnsen et al, 2000;Murphy et al, 1999).…”

mentioning

confidence: 99%

“…In this study, we used cell lines derived from the human breast adenocarcinoma MCF-7 containing either wild type (wt)-p53 or a dominant-negative variant of the p53 gene (previously shown to be resistant to microtubule-targeted drugs, Galmarini et al, 2001), as a model to investigate the possible relation between p53 expression, dynamics and composition of microtubules and drug resistance. …”

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confidence: 99%

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Drug resistance associated with loss of p53 involves extensive alterations in microtubule composition and dynamics

et al. 2003

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In the present study, we compared the dynamics and composition of microtubules in cell lines derived from the human breast adenocarcinoma MCF-7 containing either the wild-type p53 (wt-p53; MN1) or a dominant-negative variant of p53 gene (mut-p53; MDD2). Mut-p53 cells were significantly resistant to the cytotoxicity of the microtubule-targeted drugs (vinca alkaloids and taxanes), as compared with wt-p53 cells. Studies by high-resolution time-lapse fluorescence microscopy in living cells indicated that the dynamics of microtubules of mut-p53 cells were altered in complex ways and were significantly increased as compared with microtubules in wt-p53 cells. The percentage of time microtubules spent in growing and shortening phases increased significantly, their catastrophe frequency increased, and their overall dynamicity increased by 33%. In contrast, their shortening rate and the mean length shortened decreased. Cells containing mut-p53 displayed increased polymerisation of tubulin, increased protein levels of the class IV b-tubulin isotype, STOP and survivin, and reduced protein levels of class II b-tubulin isotype, MAP4 and FHIT. We conclude that p53 protein may contribute to the regulation of microtubule composition and function, and that alterations in p53 function may generate complex microtubule-associated mechanisms of resistance to tubulin-binding agents.

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Section: Cell Linesmentioning

confidence: 97%

Section: Cell Linesmentioning

confidence: 99%

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confidence: 99%

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confidence: 99%

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Drug resistance associated with loss of p53 involves extensive alterations in microtubule composition and dynamics

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Resveratrol analog (Z)‐3,5,4′‐trimethoxystilbene is a potent anti‐mitotic drug inhibiting tubulin polymerization

Schneider

Chabert

Stutzmann

et al. 2003

Intl Journal of Cancer

107

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Resveratrol (3,5,4Ј-trihydroxystilbene) is a natural polyphenol present in a variety of medicinal plants and in grapes. 1 Its function is to protect plants against pathogenic attack and environmental stress. 2,3 Because of its production by grape skin, significant amounts are found in red wines. 4 First evidence of the beneficial effects of resveratrol for human health was shown by its ability to protect against cardiovascular diseases. Resveratrol exhibits strong antioxydant activity 5,6 and inhibits platelet aggregation. 7,8 More recently, numerous cancer chemopreventive properties of resveratrol have been demonstrated . 9 -12 Ptero-stilbene, a derivative of resveratrol that is methylated at positions 3 and 5, exhibits greater antifungal activity than resveratrol. 13,14 However, this molecule is only present in very low concentrations in grapes and seems to be a minor effector of the plant defensive response. It has been shown that methylated derivatives of flavonoïds exhibit higher antiproliferative potency on cancer cells than their hydroxylated counterparts. 15 This ability may be related to the increased lipophilic properties of the methoxyflavonoids and their increased uptake through the cell membrane. On this basis we have developed a methylated derivative of resveratrol (R3: Z-3,5,4Ј-trimethoxystilbene; Fig. 1) with the aim to evaluate its antiproliferative effects on the human colon cancer cell line Caco-2.Previously resveratrol was shown to inhibit the proliferation of Caco-2 cells through the accumulation of cells in the S phase of the cell cycle and by inhibiting polyamine biosynthesis. 16 The effects of resveratrol were not cytotoxic but mainly cytostatic and reversible. In the present study, we show that R3 exerts a 100-fold higher inhibitory effect on colon cancer cell growth. The growth supressive effects induced by R3 were totally different from those observed with resveratrol and were related to a selective blockade of cells at the G2/M phase of the cell cycle, and to the disruption of the microtubule network. MATERIAL AND METHODS Synthesis of (Z)-3,5,4.ЈЈ-trimethoxystilbene (R3)The compound was synthesized by a Wittig-Horner reaction from 4-methoxybenzyl-diethyl phosphonate and 3,5-dimethoxybenzaldehyde 17,18 (Fig. 2). All reagents were commercially available and used without further purification.Melting points were measured without correction in capillary tubes on a Büchi apparatus.1 H and 13 C NMR spectra were recorded on a Bruker AC 200 MHz spectrometer. Mass spectra were established on a Fisons spectrometer at 70 eV chamber voltage, using a direct inlet tube. Silica gel 60 (40 -63 mesh, Merck Eurolab SA, Strasbourg, France) and thin layer chromatography with aluminium sheets 20 ϫ 20 cm 60 F 254 (Merck Eurolab SA, Strasbourg, France) were used. Dimethylformamide was distilled over 0.4 nm molecular sieves under reduced pressure prior to its use; 4-Methoxybenzyl alcohol (1) was purchased from Acros (Noisy le Grand, France), 3,5-dimethoxybenzaldehyde from Avocado (La Tour du Pin, France) an...

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A p21/WAF1 mutation favors the appearance of drug resistance to paclitaxel in human noncancerous epithelial mammary cells

Galmarini¹,

Bouchet²,

Audoynaud³

et al. 2006

Intl Journal of Cancer

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We investigated the mechanisms responsible for paclitaxel resistance in HME-1 cells (human mammary epithelial cells immortalized with hTERT). These cells were exposed to paclitaxel (10 pM for 7 days) and 20 cellular surviving populations (PSP) were obtained. PSP demonstrated high levels of resistance to paclitaxel cytotoxicity as compared with HME-1 cells. Activation of mdr-1 gene expression was observed in 2 PSP. Protein expression analysis using a C-terminal targeted antibody showed that 13 PSP were negative for p21/WAF1 expression after ionizing radiation (6 Gy) or doxorubicin (100 nM) treatment. Sequencing of the 3 exons of the CDKN1A gene revealed that 13 PSP contained a point mutation in exon 2. This mutation consisted in a T insertion at codon 104 leading to a premature STOP codon appearance. Mismatch amplification mutation assay and RFLP-PCR confirmed the presence of the mutation in 16 PSP. Western blot using an N-terminal targeted antibody demonstrated that the C-terminal-truncated p21/WAF1 protein (14 kDa) was indeed expressed in the 13 PSP. Our data suggest that p21/WAF1 inactivation may confer a strong resistance to paclitaxel in noncancerous breast epithelial cells harboring a p21/WAF1 mutant. ' 2006 Wiley-Liss, Inc.Key words: paclitaxel; cyclin-dependent kinase inhibitor p21; antineoplastic agent resistance Paclitaxel is one of the most effective anticancer drugs used in the clinic to treat a variety of solid tumors. In breast cancer patients, administration of paclitaxel in combination or sequentially with other chemotherapy drugs for the treatment of metastatic disease demonstrated to have more anticancer activity than previous nonpaclitaxel chemotherapy regimens. 1 Moreover, recent clinical trials also demonstrated that the use of paclitaxel as adjuvant therapy in early stage breast cancer patients helps to prevent relapse after the initial primary surgical treatment. 2,3 Despite paclitaxel increasing use, especially in therapy of curable cancers, no data are currently available on its effects on noncancerous human breast epithelial cells. This prompted us to expose hTERT-immortalized human mammary epithelial cells to a stringent selection by this agent in a 7-day drug exposure. We then analyzed the emergence of surviving cells in parallel populations. In contrast to prevalent cellular models of drug resistance derived by multiple stepwise selections, the PSP in our model have experienced a single paclitaxel treatment at concentration levels and time exposure similar than those used in clinical practice. In these experiments, mdr-1 expression was found to be positive by RT-PCR in 2 of 20 PSP (10%). Given the fact that the mdr-1-negative PSP displayed resistance to paclitaxel, we sought to determine whether other alterations existed in these PSP. Our findings indicate that mutations of the p21/WAF1 gene are involved in resistance to this agent. Material and methods ReagentsPaclitaxel was obtained from Bristol-Myers Squibb (Paris, France). Antibodies against C-terminal-p21/WAF1 (SX-118), p53 (DO7...

show abstract

Inactivation of wild-type p53 by a dominant negative mutant renders MCF-7 cells resistant to tubulin-binding agent cytotoxicity

Cited by 37 publications

References 26 publications

Drug resistance associated with loss of p53 involves extensive alterations in microtubule composition and dynamics

Drug resistance associated with loss of p53 involves extensive alterations in microtubule composition and dynamics

Resveratrol analog (Z)‐3,5,4′‐trimethoxystilbene is a potent anti‐mitotic drug inhibiting tubulin polymerization

A p21/WAF1 mutation favors the appearance of drug resistance to paclitaxel in human noncancerous epithelial mammary cells

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