Biomedical subjects

H Attalla

Publications and source records attributed to H Attalla.

5 recordsLinked to original sources

PubMed1998

2-Methoxyestradiol-induced phosphorylation of Bcl-2: uncoupling from JNK/SAPK activation.

The natural estrogen metabolite 2-methoxyestradiol (2ME) is anti-angiogenic in vivo and a strong growth inhibitor in vitro. The growth inhibition is due to mitotic arrest and apoptosis. These effects are reminiscent of those induced by taxol, and appear to be mediated by inhibition of microtubule dynamics. Here we have studied the cellular response to 2ME in regard to potential mediators of the observed cellular changes. 2ME treatment increases the insoluble polymerized fraction of cellular tubulin similar to taxol, and in contrast to the microtubule depolymerizing drugs such as colcemid and vincristine. This stabilization following 2ME treatment is accompanied by phosphorylation and inactivation of Bcl-2 increasing gradually from 2-24 hours. To study the pathway leading to Bcl-2 phosphorylation we analyzed Raf-1 and JNK/SAPK kinases, both of which have been reported to be involved in Bcl-2 inactivation. Our results indicate that Raf-1 is phosphorylated in response to 2ME, but this occurs later than Bcl-2 phosphorylation suggesting that Raf-1 is not directly phosphorylating Bcl-2. JNK/SAPK was activated rapidly after 2ME treatment. However, this activation was transient and returned to undetectable levels by 2 hours of treatment, demonstrating that JNK/SAPK is not directly phosphorylating Bcl-2. Taken together with previous results indicating that overexpression of JNK/SAPK leads to Bcl-2 phosphorylation, our results would support a model where JNK/SAPK is indirectly phosphorylating Bcl-2.

2-Methoxyestradiol↗

PubMed1998

Cytogenetic chromosomal preparations using 2-methoxyestradiol.

The antimitotic drug 2-methoxyestradiol is an end metabolite of catechol estrogens. In vivo, it arrests cells in mitosis by interfering with the dynamics of the mitotic spindle without disrupting tubulin formation. It has a mitotic index similar to that of Colcemid in different cell lines. Here we report that 2-methoxyestradiol can be used for making cytogenetic preparations of comparable quality to that of colcemid. In addition, 2-methoxyestradiol is devoid of the toxicity associated with Colcemid, which may make 2-methoxyestradiol useful in slowly growing samples often found in primary solid tumor cultures where a sufficient number of mitotic cells is difficult to obtain.

2-Methoxyestradiol↗

PubMed1997

2,6-Bis((3,4-dihydroxyphenyl)-methylene)cyclohexanone (BDHPC)-induced apoptosis and p53-independent growth inhibition: synergism with genistein.

Estrogen binds to two classes of proteins in the cells, the high-affinity estrogen receptor (ER) as well as a low affinity estrogen type II binding site (EBS-II). Methyl p-hydroxyphenyllactate (MeHPLA) is an endogenous ligand for EBS-II. Binding of MeHPLA to EBS-II has a growth regulatory effect in estrogen-responsive cells, and levels of MeHPLA are decreased in breast cancer due to degradation by a specific esterase. 2,6-bis((3, 4-dihydroxyphenyl)-methylene) cyclohexanone (BDHPC) is an esterase-resistant analogue of MeHPLA which binds irreversibly to EBS-II and inhibits growth of breast cancer cells. In the present study, we analyzed the mechanism of growth inhibition by BDHPC. Treatment with BDHPC resulted in accumulation of cells in G1 phase and apoptosis. The G1 accumulation was not dependent on a functional p53 gene. The G1-specific growth inhibition by BDHPC was found to act synergistically with the G2/M-specific inhibition induced by the tyrosine kinase inhibitor genistein, suggesting this drug combination could be effectively used in cancer treatment.

Antineoplastic Agents↗

PubMed1996

2-Methoxyestradiol arrests cells in mitosis without depolymerizing tubulin.

The endogenous estrogen metabolite 2-methoxyestradiol (2-MeOE2) suppresses experimental tumor growth in vivo and inhibits angiogenesis activity in vitro. Moreover, 2-MeOE2 has been observed to block mitosis in cell cultures. As high concentrations of 2-MeOE2 prevent microtubule assembly in vitro, the mitotic arrest has been attributed to inhibition of tubulin polymerization. Here we report that concentrations of 2-MeOE2 that cause complete metaphasal arrest do not inhibit the assembly of mitotic spindles. In contrast to the chromosomal dispersal seen in cells arrested by the tubulin depolymerizing drug colcemid, the chromosomes of cells treated with 2-MeOE2 remained in the metaphasal plate indicating a functional defect of the mitotic spindle. The 2-MeOE2 arrest resembles those induced by compounds affecting microtubule dynamics such as taxol and vinblastine. The 2-MeOE2 block is also similar to that induced by several anti-calmodulin agents. Given that metaphase to anaphase transition is a calmodulin-dependent step and our observation that 2-MeOE2 inhibits calmodulin activity in vitro, we suggest that the 2-MeOE2 metaphasal arrest may occur via inhibition of calmodulin.

2-Methoxyestradiol↗

PubMed1995

Lignan and isoflavonoid conjugates in human urine.

Lignans and isoflavonoids are two groups of diphenolic phytoestrogens of plant origin which have gained increasing interest because of their possible cancer protective properties. High excretion of these compounds occur in populations at low risk of breast, prostate and colon cancer consuming either high amounts of whole-grain (lignans and some isoflavonoids) or soy products (isoflavonoids and some lignans). We determined the pattern of conjugation of the phytoestrogens in four urine samples from vegetarian or semivegetarian women and in two samples from men. Seven compounds were investigated: enterodiol, enterolactone, matairesinol, diadzein, equol, genistein and O-desmethylangolensin. The fractions quantified are the free fraction, mono- and disulfate, as well as the mono-, di- and sulfoglucuronide fractions. For the fractionation and purification we used ion-exchange chromatography and the determination of the concentrations of each compound in all fractions was done by isotope dilution gas chromatography-mass spectrometry (GLC-MS) using deuterated internal standards of all diphenols. More than 60% of all compounds determined, occurred in the monoglucuronide fraction. Daidzein, enterodiol and equol are excreted to a relatively high extent as sulfoglucuronides and genistein as diglucuronide. We conclude that the general pattern of lignan and isoflavonoid conjugates in urine is similar to that of endogenous estrogens.

Estrogens↗