Trichostatin A enhances acetylation as well as protein stability of ERα through induction of p300 protein

Introduction  

Trichostatin A (TSA) is a well-characterized histone deacetylase (HDAC) inhibitor. TSA modifies the balance between HDAC and histone acetyltransferase activities that is important in chromatin remodeling and gene expression. Although several previous studies have demonstrated the role of TSA in regulation of estrogen receptor alpha (ERα), the precise mechanism by which TSA affects ERα activity remains unclear.     

Antisense depletion of RIα subunit of protein kinase A induces apoptosis and growth arrest in human breast cancer cells

In recent years, several laboratories have explored the possibility of using antisense oligodeoxynucleotides for specific manipulation of gene expression leading to cancer treatment. The enhanced expression of the RI subunit of cyclic AMP-dependent protein kinase type I (PKA-I) has been correlated with cancer cell growth. In the present study, the effects of an antisense oligodeoxynucleotide targeted against RI subunit of PKA-I on growth inhibition and apoptosis in MDA-MB-231 human breast cancer cells were investigated. The growth inhibitory effects of RI antisense oligodeoxynucleotide correlated with a decrease in the RI mRNA and protein levels. The growth inhibition was accompanied by changes in the cell cycle phase distribution, cell morpbology, cleavage of poly (ADP-ribose) polymerase (PARP), and appearance of apoptotic nuclei. By comparison, mismatched control oligodeoxynucleotide had no effect. On the basis of these results, it can be suggested that the RI antisense oligodeoxynucleotide, which efficiently depletes the growth stimulatory RI and induces apoptosis/differentiation, could be used as a therapeutic agent for breast cancer treatment.     

ER−/ER+ breast cancer cell lines exhibited different resistance to paclitaxel through pulse selection

In this study, we established two PTX-resistant breast cancer cell lines, 231 TIM10 and MCF-7 TIM10, from ER-negative MDA-MB-231 cells and ER-positive MCF-7 cells by pulse selection, respectively. We found that 231 TIM10 variants acquired higher drug resistance than MCF-7 TIM10 variants by the pulse selection, although ER-positive MCF-7 cells were not as sensitive as ER-negative MDA-MB-231 to the initial pulses with PTX. 231 TIM10 had 11.9-fold greater resistance (RI = 11.9) than the parental MDA-MB-231 cells, while MCF-7 TIM10 got 5.5-fold resistance (RI = 5.5) when compared with the parental MCF-7 cells. In the presence of 5nM PTX, 231 TIM10 cells formed colonies, but no colony formed when MCF-7 TIM10 cells were cultured in the same condition. These data have two implications. First, the ER expression state might be an important determinant for the response of breast cancer cells to paclitaxel treatment. Second, ER-negative and ER-positive breast cancer cells develop drug-resistance phenotype with distinctive mechanisms. Our work not only established useful models for studying the paclitaxel resistance but also provides interesting clues to understand the mechanisms underlying the drug resistance of ER-negative and ER-positive breast cancer cells.     

Transactivation of ERα by Rosiglitazone induces proliferation in breast cancer cells

In the present study, we demonstrate that Rosiglitazone (Rosi), a thiazolidinedione and PPARγ agonist, induces ERE (Estrogen Receptor Response Element) reporter activity, pS2 (an endogenous ER gene target) expression, and proliferation of ER positive breast cancer (MCF-7) cells. By performing a dose–response assay, we determined that high concentrations of Rosi inhibit proliferation, while low concentrations of Rosi induce proliferation. Using the anti-estrogen ICI, ER negative breast cancer (MDA-MB-231) cells, and a prostate cancer cell line (22Rv1) deficient in both ERα and PPARγ, we determined that Rosiglitazone-induced ERE reporter activation and proliferation is through an ERα dependent mechanism. Rosiglitazone-induced ERE activation is also dependent on activation of the Extracellular Signal-Regulated Kinase–Mitogen Activated Protein Kinase (ERK–MAPK) pathway, since it is inhibited by co-treatment with U0126, a specific inhibitor of this pathway. We also demonstrate that when ERα and PPARγ are both present, they compete for Rosi, inhibiting each others transactivation. To begin to unravel the pharmacological mechanism of Rosi-induced ER activation, sub-maximally effective concentrations of E₂ were used in combination with increasing concentrations of Rosi in luciferase reporter assays. From these assays it appears that E₂ and Rosi both activate ERα via similar pharmacological mechanisms. Furthermore sub-maximally effective concentrations of E₂ and Rosi additively increase both ERE reporter activity and MCF-7 cell proliferation. The results of this study may have clinical relevancy for Rosi’s use both as an anti-diabetic in post-menopausal women and as an anti-cancer drug in women with ER positive breast cancer     

Human breast cancer-associated fibroblasts enhance cancer cell proliferation through increased TGF-α cleavage by ADAM17

We demonstrate here increased expression of ADAM17 protein in cancer-associated fibroblasts (CAFs) extracted from human breast carcinomas compared with donor-matched normal fibroblasts, and TGF-α secretion positively correlates with ADAM17 expression in these cells. In SK-BR-3 cells co-cultured with CAFs, CAF-secreted TGF-α promotes cell proliferation by activation of EGFR, Akt, and ERK, but it does not promote cell migration. Furthermore, anti-TGF-α neutralizing antibodies antagonize the CAF-dependent increase in proliferation and activation of EGFR, Akt and ERK. Thus, pharmacologic inhibition of ADAM17 and TGF-α may have therapeutic potential for the treatment of breast cancer when fibroblast-directed therapy is considered.     

Chronic acidosis rewires cancer cell metabolism through PPARα signaling

The mechanisms linking tumor microenvironment acidosis to disease progression are not understood. Here, we used mammary, pancreatic, and colon cancer cells to show that adaptation to growth at an extracellular pH (pH_e) mimicking acidic tumor niches is associated with upregulated net acid extrusion capacity and elevated intracellular pH at physiological pH_e, but not at acidic pH_e. Using metabolic profiling, shotgun lipidomics, imaging and biochemical analyses, we show that the acid adaptation-induced phenotype is characterized by a shift toward oxidative metabolism, increased lipid droplet-, triacylglycerol-, peroxisome content and mitochondrial hyperfusion. Peroxisome proliferator-activated receptor-α (PPARA, PPARα) expression and activity are upregulated, at least in part by increased fatty acid uptake. PPARα upregulates genes driving increased mitochondrial and peroxisomal mass and β-oxidation capacity, including mitochondrial lipid import proteins CPT1A, CPT2 and SLC25A20, electron transport chain components, peroxisomal proteins PEX11A and ACOX1, and thioredoxin-interacting protein (TXNIP), a negative regulator of glycolysis. This endows acid-adapted cancer cells with increased capacity for utilizing fatty acids for metabolic needs, while limiting glycolysis. As a consequence, the acid-adapted cells exhibit increased sensitivity to PPARα inhibition. We conclude that PPARα is a key upstream regulator of metabolic changes favoring cancer cell survival in acidic tumor niches.     

Inhibition of novel protein kinase Cɛ augments TRAIL-induced cell death in A549 lung cancer cells

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) has great potential for cancer treatment since it provokes cell death in most tumor cells while leaving most normal cells unscathed. Some cancers, however, show resistance to TRAIL, indicating that TRAIL alone may be insufficient for cancer therapy. Here we studied whether the apoptotic susceptibility of A549 non-small cell lung cancer cells could be modulated by inhibiting protein kinase C (PKC). We show that an inhibitor with preference for novel PKC isozymes, NPC 15437, significantly augmented TRAIL sensitivity of A549 cells, as judged by assessing cell death and mitochondrial membrane potential. Likewise, NPC 15437 also significantly potentiated the responsiveness of DAOY medulloblastoma cells to TRAIL. In contrast, an inhibitor with preference for conventional PKC isozymes, Gö6976, did not augment TRAIL sensitivity of A549 cells. To further specify the PKC isozyme responsible for TRAIL sensitization, we used a peptide inhibitor with selectivity for the novel PKC isozyme ?, myr-PKC? V1-2. The inhibition of PKC? resulted in a significant amplification of the cytotoxic activity of TRAIL in A549 cells. Altogether, our study provides evidence for a considerable role of PKC? in the apoptotic responsiveness of A549 lung cancer cells, and possibly other malignancies, to TRAIL.     

Association of promoter methylation of ERα and ERβ with sporadic breast cancer—a study from North India

Methylations in estrogen receptor (ER) α and ERβ are known to be involved in the pathogenesis of breast cancer. Here, we explore the role of promoter methylation of estrogen receptors, ERα and ERβ, in sporadic breast cancer cases from a North Indian population. To this end, association between ERα and ERβ methylation status along with different clinicopathological parameters and its correlation with protein expression was examined. Four hundred eighty paired breast cancer tissue samples and adjacent normal controls from 240 sporadic breast cancer patients were included, and their clinical and demographic profiles were recorded. ERα and ERβ methylation was determined by methylation-specific polymerase (MSP) chain reaction. Our findings demonstrate that methylation of ERα and ERβ occurs in high frequency and appears to be a mechanism of gene silencing in our population. Furthermore, on performing stratified analysis, we observed strong associations between ERα/ERβ methylation and ER, PR, and HER2 status, tumor size, clinical stage, and triple negative tumors. Thus, our study not only highlights the role of ERα/ERβ methylation in breast cancer but also suggests the ERα/ERβ methylation pattern as a biomarker for assessing breast cancer risk.     

A novel taspine derivative, HMQ1611, inhibits breast cancer cell growth via estrogen receptor α and EGF receptor signaling pathways

Breast cancer is a common cancer with a leading cause of cancer mortality in women. Currently, the chemotherapy for breast cancer is underdeveloped. Here, we report a novel taspine derivative, HMQ1611, which has anticancer effects using in vitro and in vivo breast cancer models. HMQ1611 reduced cancer cell proliferation in four human breast cancer cell lines including MDA-MB-231, SK-BR-3, ZR-75-30, and MCF-7. HMQ1611 more potently reduced growth of estrogen receptor α (ERα)-positive breast cancer cells (ZR-75-30 and MCF-7) than ERα-negative cells (MDA-MB-231 and SK-BR-3). Moreover, HMQ1611 arrested breast cancer cell cycle at S-phase. In vivo tumor xenograft model, treatment of HMQ1611 significantly reduced tumor size and weight compared with vehicles. We also found that HMQ1611 reduced ERα expression and inhibited membrane ERα-mediated mitogen-activated protein kinase (MAPK) signaling following the stimulation of cells with estrogen. Knockdown of ERα by siRNA transfection in ZR-75-30 cells attenuated HMQ1611 effects. In contrast, overexpression of ERα in MDA-MB-231 cells enhanced HMQ1611 effects, suggesting that ERα pathway mediated HMQ1611's inhibition of breast cancer cell growth in ERα-positive breast cancer. HMQ1611 also reduced phosphorylation of EGF receptor (EGFR) and its downstream signaling players extracellular signal-regulated kinase (ERK)1/2 and AKT activation both in ZR-75-30 and MDA-MB-231 cells. These results showed that the novel compound HMQ1611 had anticancer effects, and partially via ERα and/or EGFR signaling pathways, suggesting that HMQ1611 may be a potential novel candidate for human breast cancer intervention.     

Disruption of ERα signalling pathway by PPARγ agonists: evidences of PPARγ-independent events in two hormone-dependent breast cancer cell lines

Peroxisome proliferator-activated receptor γ (PPARγ) is a nuclear receptor that can be activated by natural ligands such as 15-deoxy-delta(12,14)-prostaglandin J2 (15d-PGJ(2)) as well as synthetic drugs such as thiazolidinediones. The treatment of human breast cancer cell lines with PPARγ agonists is known to have antiproliferative effects but the role of PPARγ activation in the process remains unclear. In the present study, we investigated the effects of four PPARγ agonists, Rosiglitazone (RGZ), Ciglitazone (CGZ), Troglitazone (TGZ) and the natural agonist 15d-PGJ(2), on estrogen receptor alpha (ERα) signalling pathway in two hormone-dependent breast cancer cell lines, MCF-7 and ZR-75-1. In both of them, TGZ, CGZ and 15d-PGJ(2) induced an inhibition of ERα signalling associated with the proteasomal degradation of ERα. ZR-75-1 cells were more sensitive than MCF-7 cells to these compounds. Treatments that induced ERα degradation inhibited cell proliferation after 24 h. In contrast, 24 h exposure to RGZ, the most potent activator of PPARγ disrupted neither ERα signalling nor cell proliferation. 9-cis retinoic acid never potentiated the proteasomal degradation of ERα. PPARγ antagonists (T0070907, BADGE and GW 9662) did not block the proteolysis of ERα in MCF-7 and ZR-75-1 cells treated with TGZ. ERα proteolysis still occurred in case of PPARγ silencing as well as in case of treatment with the PPARγ-inactive compound Δ2-TGZ, demonstrating a PPARγ-independent mechanism. The use of thiazolidinedione derivatives able to trigger ERα degradation by a PPARγ-independent pathway could be an interesting tool for breast cancer therapy.