Low concentrations of 17ß-estradiol exacerbate tamoxifen resistance in breast cancer treatment through membrane estrogen receptor-mediated signaling pathways.

The present study aims to discover the influences of tamoxifen and 17ß-estradiol (E2) on tamoxifen-resistant (TamR) patients in vitro. Herein, we established a stabilized TamR MCF-7 cell line at 1 µM via gradient concentrations of tamoxifen cultivation. The expression changes of four ER subtypes (ERα66, ERß, ERα36 and GPR30) were found to bring about tamoxifen resistance. Moreover, the generation of tamoxifen resistance involved in apoptosis escape via a reactive oxygen species-regulated p53 signaling pathway. Interestingly, E2 at environmental concentrations (0.1-10 nM) could activate the expression of both ERα36 and GPR30, and then stimulate the phosphorylation of ERK1/2 and Akt, resulting in cell growth promotion. Cell migration and invasion promotion, apoptosis inhibition, and cell cycle G1-S progression are involved in such proliferative effects. Conversely, the application of specific antagonists of ERα36 and GPR30 could restore tamoxifen's sensitivity as well as partially offset E2-mediated proliferation. In short, overexpression of ERα36 and GPR30 not only ablate tamoxifen responsiveness but also could promote tumor progression of TamR breast cancer under estrogen conditions. These results provided novel insights into underlying mechanisms of tamoxifen resistance and the negative effects of steroid estrogens at environmental concentrations on TamR MCF-7 cells, thus generating new thoughts for future management of ER-positive breast cancer.

The estrogenic proliferative effects of two alkylphenols and a preliminary mechanism exploration in MCF-7 breast cancer cells.

Bisphenol A (BPA) and 4-cumylphenol (4-CP), as estrogen-like chemicals, are ubiquitous in the environment media and associated with the occurrence and development of hormone-dependent tumors. However, the combinatorial effects of these two structurally similar alkylphenols are not well informed. In the present study, the classic breast cancer cell line MCF-7 was used as in vitro model to estimate the estrogenic proliferative effects of BPA and 4-CP. MTT assay, reactive oxygen species, cell apoptosis, cell cycle, and real-time fluorescent quantitative Step One Plus Real-time PCR System (Applied Biosystems, CA, USA) were applied to explore their proliferative mechanisms. MTT results showed that both BPA and 4-CP ranging from 10-9 to 10-5 M stimulated cell proliferation in a nonmonotonic dose-response manner. Along with the proliferative effects, cell cycle was progressed from G0/G1 to S and G2/M phase. Meanwhile, the expression levels of ERα, pS2, and Bcl-2 mRNA were also upregulated. In contrast, 4-CP and BPA at high dose (10-4 M) obviously displayed antiproliferative effects in MCF-7 cells via inducing cell apoptosis and blocking cell cycle in G0/G1 phase. As expected, the relative expression levels of ERα, pS2, and Bcl-2 mRNA were decreased, whereas Bax mRNA was increased. Interestingly, the proliferative or antiproliferative effects of 4-CP were higher than that of BPA. Moreover, coexposure of lower concentrations BPA and 4-CP significantly induced cell proliferation in a synergistic manner. These findings indicated that the potential environmental risks of coexposure of BPA and 4-CP were greater than either of them.

Quercetin exerts bidirectional regulation effects on the efficacy of tamoxifen in estrogen receptor-positive breast cancer therapy: An in vitro study.

Tamoxifen was widely applied in the therapy of estrogen receptor (ER)-positive breast cancer. With the purpose of determining the potential impacts of quercetin on its effectiveness, MCF-7 cells were selected as the in vitro model and several cellular biological behaviors (ie, cell proliferation, migration, invasion, cycle, apoptosis, and oxidative stress) were investigated. As results, quercetin showed contrasting dose-response to cellular behaviors dependent on the ROS-regulated p53 signaling pathways. In detail, quercetin promoted cell proliferation and inhibited cell apoptosis at low concentrations, whereas high-concentration resulted in apoptosis induction. Moreover, quercetin at a low concentration significantly inhibited tamoxifen-induced antiproliferation in MCF-7 cells, whereas high concentrations enhanced cell apoptosis in a synergetic manner. The real-time quantitative polymerase chain reaction analysis further implied that quercetin exerted its dual roles in tamoxifen-induced antiproliferative effects by regulated the gene expression involved in cell metastasis, cycle, and apoptosis through the ER pathways. Our present study provides a considerable support to the combination of quercetin and tamoxifen on human ER-positive breast carcinoma management.

How does the neurotoxin ß-N-methylamino-L-alanine exist in biological matrices and cause toxicity?

The neurotoxin ß-N-methylamino-L-alanine (BMAA) has been deemed as a risk factor for some neurodegenerative diseases such as amyotrophic lateral sclerosis/parkinsonism dementia complex (ALS/PDC). This possible link has been proved in some primate models and cell cultures with the appearance that BMAA exposure can cause excitotoxicity, formation of protein aggregates, and/or oxidative stress. The neurotoxin BMAA extensively exists in the environment and can be transferred through the food web to human beings. In this review, the occurrence, toxicological mechanisms, and characteristics of BMAA were comprehensively summarized, and proteins and peptides were speculated as its possible binding substances in biological matrices. It is difficult to compare the published data from previous studies due to the inconsistent analytical methods and components of BMAA. The binding characteristics of BMAA should be focused on to improve our understanding of its health risk to human health in the future.

ß-N-methylamino-L-alanine production, photosynthesis and transcriptional expression in a possible mutation strain and a wild strain of Thalassiosira minima.

The neurotoxin ß-N-methylamino-L-alanine (BMAA) produced by marine diatoms has been implicated as an important environmental trigger of neurodegenerative diseases in humans. However, the biosynthesis mechanism of BMAA in marine diatoms is still unknown. In the present study, the strain of diatom Thalassiosira minima almost lost the biosynthesis ability for BMAA after a long-term subculture in our laboratory. The production of BMAA-containing proteins in the mutant strain of T. minima reduced to 18.2 % of that in the wild strain, meanwhile the cell size decreased but pigment content increased in the mutant strain. Take consideration of our previous transcriptional data on the mixed diatom and cyanobacterium cultures, the current transcriptome analysis showed four identical and highly correlated KEGG pathways associated with the accumulation of misfolded proteins in diatom, including ribosome, proteasome, SNARE interactions in vesicle transport, and protein processing in the endoplasmic reticulum. Analysis of amino acids and transcriptional information suggested that amino acid synthesis and degradation are associated with the biosynthesis of BMAA-containing proteins. In addition, a reduction in the precision of ubiquitination-mediated protein hydrolysis and vesicular transport by the COPII system will exacerbate the accumulation of BMAA-containing proteins in diatoms.

Cell cycle of microalga Isochrysis galbana arrested by neurotoxin ß-N-methylamino-l-alanine and corresponding molecular mechanisms.

The phycotoxin ß-N-methylamino-l-alanine (BMAA) has attracted attention due to its risks to marine organisms and human health. In this study, approximately 85 % of synchronized cells of the marine microalga Isochrysis galbana were arrested at the cell cycle G1 phase by BMAA at 6.5 µM for a 24-h exposure. The concentration of chlorophyll a (Chl a) gradually decreased, while the maximum quantum yield of PSII (Fv/Fm), the maximum relative electron transport rate (rETRmax), light utilization efficiency (α) and half-saturated light irradiance (Ik) reduced early and recovered gradually in I. galbana exposed to BMAA in 96-h batch cultures. Transcriptional expression of I. galbana analyzed at 10, 12, and 16 h disclosed multiple mechanisms of BMAA to suppress the microalgal growth. Production of ammonia and glutamate was limited by the down-regulation of nitrate transporters, glutamate synthase, glutamine synthetase, cyanate hydrolase, and formamidase. Diverse extrinsic proteins related to PSII, PSI, cytochrome b6f complex, and ATPase were influenced by BMAA at transcriptional level. Suppression of the DNA replication and mismatch repair pathways increased the accumulation of misfolded proteins, which was reflected by the up-regulated expression of proteasome to accelerate proteolysis. This study improves our understanding of the chemical ecology impacts of BMAA in marine ecosystems.

Double-dose responses of Scenedesmus capricornus microalgae exposed to humic acid.

Dissolved organic matter (DOM) has been found to attenuate the ecotoxicity of various environmental pollutants, but research on its own toxic effects in aquatic ecosystems has been very limited. Herein, the toxic effects of humic acid (HA), a represent DOM typically found in natural waters, on the freshwater alga Scenedesmus capricornus were investigated. As result, HA exerted a double-dose effect on the growth of Scenedesmus capricornus. At HA concentrations below 2.0 mgC/L, the growth of Scenedesmus capricornus was slightly promoted, as was the synthesis of chlorophyll and macromolecules in the algae. Moreover, S. capricornus can maintain its growth by secreting fulvic acid as a nutrient carbon source. However, the growth of Scenedesmus capricornus was significantly inhibited when HA was beyond 2.0 mgC/L. The main mechanisms of humic acid's toxicity were membrane damage and oxidative stress. Particularly, when the oxidative stress exceeds the algae's carrying capacity, the synthesis of EPS is greatly inhibited and HA damage results. Taken together, DOM may have both positive and negative effects on aquatic ecosystems.

Modified humic acids mediate efficient mineralization in a photo-bio-electro-Fenton process.

Humic acids (HA) are common mediators in redox reactions in the aquatic environment. The structures and properties of HA are greatly influenced by environmental factors such as external electrons. In this study, qualitative changes in electron-modified HA and the underlying mechanisms were reported, which not only contribute to understanding the fate of HA and their impact on organic pollutants, but could facilitate their potential use for water purification. The photochemical activity and electron-donating capacity of HA were improved due to the increase of phenolic and carboxyl components via the reduction modification by electrons, creating a novel and efficient photo-bio-electro-Fenton system mediated by HA under neutral conditions without the use of hydrogen peroxide (H2O2). The in-situ continuous production of H2O2 ensured an adequate supply of hydroxyl radicals in this coupled system, achieving mineralization (92%) of HA and 17α-ethinylestradiol (EE2), a common synthetic estrogen with high estrogenic potency. Two degradation pathways with five degradation intermediates of EE2 were identified in our study. Effluents from the coupled system showed decreased endocrine-disrupting activity. Our findings demonstrated a new approach for the in-situ modification and potential use of HA for water treatment and particularly the concurrent degradation of HA and organic pollutants through a photo-bioelectrochemical system mediated by HA.

A critical review on the applications and potential risks of emerging MoS2 nanomaterials.

Molybdenum disulfide (MoS2) nanomaterials have been widely used in various fields such as energy store and transformation, environment protection, and biomedicine due to their unique physicochemical properties. Unfortunately, such large-scale production and use of MoS2 nanomaterials would inevitably release into the environmental system and then potentially increase the risks of wildlife/ecosystem and human beings as well. In this review, we first introduce the physicochemichemical properties, synthetic methods and environmental behaviors of MoS2 nanomaterials and their typical functionalized materials, then summarize their environmental and biomedical applications, next assess their potential health risks, covering in vivo and in vitro studies, along with the underlying toxicological mechanisms, and last point out some special phenomena about the balance between applications and potential risks. This review aims to provide guidance for harm predication induced by MoS2 nanomaterials and to suggest prevention measures based on the recent research progress of MoS2' applications and exerting toxicological data.

17ß-estradiol at low concentrations attenuates the efficacy of tamoxifen in breast cancer therapy.

Tamoxifen has been applied widely in the treatment of estrogen receptor (ER)-positive breast cancer. The impact of low concentrations of 17ß-estradiol (E2) (a pervasive environmental pollutant) on its effectiveness was studied in vitro using an MCF-7 cell line. Cell proliferation, migration, invasion, and apoptosis were studied along with cell cycle progression, reactive oxygen species generation and mitochondrial membrane potentials repression. The signaling pathways involved were identified. Typical concentrations of E2 in the environment (10-10 to 10-8 M) were observed to promote cell growth and protect MCF-7 cells from tamoxifen's cytotoxicity. Cell migration, invasion, cell cycle progression and apoptosis all involved in reducing tamoxifen's cytotoxicity. E2 at environmental concentrations induced PI3K/Akt and MAPK/ERK signal transduction through the estrogen receptor pathways to affect cell proliferation. Taken together, the results explain how E2 in the environment may attenuate the efficacy of tamoxifen in ER-positive breast cancer therapy. They provide considerable support for E2's adverse effects on human health and cancer management.

Combinatorial anti-proliferative effects of tamoxifen and naringenin: The role of four estrogen receptor subtypes.

Breast cancer is the most diagnosed diseases and the second-leading cause of death in females, among which the estrogen receptor positive (ER+) patients are more common of all cases. In present study, we selected MCF-7 as an in vitro model and investigated the combinatorial anti-proliferative effects of tamoxifen and naringenin on ER+ breast cancer, and then explored the potential mechanisms involved in mitochondrial dysfunction and oxidative stress mediated by several estrogen receptor subtypes. Six assessment endpoints including cell viability, cell migration, cell cycle, cell apoptosis, mRNA, and protein expression were estimated. Tamoxifen and naringenin were shown to inhibit the cell growth of MCF-7 cells at higher concentrations, and co-exposure with them significantly inhibited cell proliferation in an additive manner. Results from a wound healing assay indicated that the combined treatment of tamoxifen and naringenin markedly suppressed cell migration compared with the single exposure by downregulating the expression of MMP-9 and MMP-2. Flow cytometry analysis revealed that the combined treatment of tamoxifen and naringenin blocked cell cycle in G2/M phase through suppressing the transcription of cell cycle regulation proteins. Simultaneously, co-treatment with them also induced cell apoptosis by regulating the expression of mitochondrial apoptotic proteins as well as by simulating the production of reactive oxygen species (ROS). Real-time quantitative polymerase chain reaction (RT-qPCR) and western blotting (WB) analysis results further demonstrated that the two nuclear estrogen receptors-ERα66 and ERß, as well as the two membrane estrogen receptors-ERα36 and GPR30 were downregulated when cells were exposed to single tamoxifen, whereas naringenin treatment group not only downregulated the expression of ERα66 and GPR30 but also upregulated ERß and ERα36. Interestingly, co-treatment group resulted in significant downregulation of ERα66, ERα36 and GPR30 but upregulation of ERß. Taken together, co-treatment of tamoxifen and naringenin could inhibit cell proliferation more effectively in ER+ breast cancer cells, which was associated with a modulation of the expression levels of several estrogen receptors.