Prognostic implications of alpha-fetoprotein and C-reactive protein elevation in hepatocellular carcinoma following resection (PACE): a large cohort study of 2770 patients.

BACKGROUND: Routine clinical staging for hepatocellular carcinoma (HCC) incorporates liver function, general health, and tumor morphology. Further refinement of prognostic assessments and treatment decisions may benefit from the inclusion of tumor biological marker alpha-fetoprotein (AFP) and systemic inflammation indicator C-reactive protein (CRP). METHODS: Data from a multicenter cohort of 2770 HCC patients undergoing hepatectomy were analyzed. We developed the PACE risk score (Prognostic implications of AFP and CRP Elevation) after initially assessing preoperative AFP and CRP's prognostic value. Subgroup analyzes were performed in BCLC cohorts A and B using multivariable Cox analysis to evaluate the prognostic stratification ability of the PACE risk score and its complementary utility for BCLC staging. RESULTS: Preoperative AFP ≥ 400ng/mL and CRP ≥ 10 mg/L emerged as independent predictors of poorer prognosis in HCC patients who underwent hepatectomy, leading to the creation of the PACE risk score. PACE risk score stratified patients into low, intermediate, and high-risk groups with cumulative 5-year overall (OS) and recurrence-free survival (RFS) rates of 59.6%/44.9%, 43.9%/38.4%, and 20.6%/18.0% respectively (all P < 0.001). Increased PACE risk scores correlated significantly with early recurrence and extrahepatic metastases frequency (all P < 0.001). The multivariable analysis identified intermediate and high-risk PACE scores as independently correlating with poor postoperative OS and RFS. Furthermore, the PACE risk score proficiently stratified the prognosis of BCLC stages A and B patients, with multivariable analyses demonstrating it as an independent prognostic determinant for both stages. CONCLUSION: The PACE risk score serves as an effective tool for postoperative risk stratification, potentially supplementing the BCLC staging system.

Chrysophanol localizes in mitochondria to promote cell death through upregulation of mitochondrial cyclophilin D in HepG2 cells.

Objective: Chrysophanol (Chry) displays potent anticancer activity in human cancer cells and animal models, but the cellular targets of Chry have not been fully defined. Herein, we speculated whether mitochondria were a target involved in Chry-induced cytotoxicity. Methods: Human liver cancer cell line HepG2 was incubated. The cytotoxicity was evaluated by MTT assay. Mitochondria localization was evaluated by a confocal microscopy. Mitochondrial membrane potential ΔΨm was detected by TMRE staining and determined by the flow cytometer. The levels of ATP, mitochondrial superoxide anions, and GSH/GSSG were determined according to the assay kits. The apoptosis were evaluated through Hoechst33342/PI and Annexin V/PI staining, respectively. The expression of cyclophilin D (CyPD) was determined by immunoblot method, and the interaction between CyPD and Chry was analyzed by molecule docking procedure. Results: Chry itself mainly localized in mitochondria to cause mitochondrial dysfunction and cell death in HepG2 cells. As regard to the mechanism, cyclosporin A as the inhibitor for the formation of mitochondrial permeability transition pore (mPTP) moderately suppressed cell death, indicating mPTP involved in the process of cell death. Further, Chry enhanced the protein expression of Cyclophilin D (CyPD) which is a molecular componentry and a modulator of mPTP, while antioxidant N-acetyl-L-cysteine inhibited the expression of CyPD. Molecule docking procedure disclosed two hydrogen-bonds existed in CyPD-Chry complex with -11.94 kal/mol of the binding affinity value. Besides, the mtDNA-deficient HepG2-ρ0 cells were much resistant to Chry-induced cell death, indicating mtDNA at least partly participated in cell death. A combination of Chry and VP-16 produced the synergism effect toward cell viability and ΔΨm, while Chry combined with Cis-Pt elicited the antagonism effect. Conclusion: Taken together, enrichment in mitochondria and actions on mPTP, CyPD and mtDNA provides an insight into the anticancer mechanism of Chry. The combination therapy for Chry with clinical drugs may deserve to further explore.

[Effect on clinical symptoms and mechanism of chloasma in premenopausal women with liver qi stagnation treated with acupoint catgut embedding].

OBJECTIVE: To explore the therapeutic effect and mechanism of acupoint catgut embedding for chloasma in premenopausal women with liver qi stagnation. METHODS: A total of 92 patients of chloasma in premenopausal women with liver qi stagnation were randomized into an observation group and a control group, 46 cases in each one.In the observation group, acupoint catgut embedding was applied at Ganshu (BL 18), Pishu (BL 20), Sanyinjiao (SP 6), also the surrounding acupuncture was performed at facial part, once a week. In the control group, vitamin C (200 mg per time, 3 times a day) and vitamin E (100 mg per time, once a day) were prescribed for oral administration. Both of the two groups were given treatment for 12 weeks. The score of chloasma area and severity index (MASI), the serum levels of follicular stimulating hormone (FSH), leuteinizing hormone (LH) and estradiol (E2) before and after treatment were observed in the two groups, and the clinical effect of the two groups were compared after 3 months of treatment. RESULTS: Finally, 44 cases in the observation group and 43 cases in the control group completed the study. The effective rate in the observation group was 88.6% (39/44), which was higher than 55.8% (24/43) in the control group (P<0.01). Compared before treatment, the MASI scores after treatment were reduced in the two groups (P<0.01, P<0.05), the MASI score in follow-up in the observation group was reduced as compared with after treatment (P<0.01), and the MASI scores after treatment and in follow-up in the observation group were lower than the control group (P<0.01). Compared before treatment, the levels of FSH and LH in follow-up were reduced (P<0.01), there was no significant difference between before treatment and in follow-up in the level of E2 in the observation group (P>0.05). There was no significant difference between before treatment and in follow-up in the levels of FSH, LH and E2 in the control group (P>0.05). In follow-up, the levels of FSH and LH in the observation group were lower than the control group (P<0.01, P<0.05), there was no significant difference in the level of E2 between the two groups (P>0.05). CONCLUSION: Acupoint catgut embedding can improve the clinical symptoms of chloasma in premenopausal women with liver qi stagnation, the mechanism may be related to regulate the serum level of sex hormone to reduce high sensitivity of melanocytes to estrogen.

[Signaling pathways controlling skeletal muscle mass].

The skeletal muscle mass accounts for more than 40% of the body weight of healthy adults. The skeletal muscle not only plays an important role in physical activities but also affects the function of other organs as a secretory organ secreting multiple muscle factors. Therefore, it is important to maintain the normal quantity and function of skeletal muscle. Skeletal muscle mass is the basis of skeletal muscle function and is often affected by many factors such as exercise and disease. Resistance exercise training induces increased protein synthesis in skeletal muscle cells, while limb disuse, chronic obstructive pulmonary disease, heart failure, chronic kidney disease, cachexia, Duchenne muscular dystrophy and many other pathological conditions lead to decreased protein synthesis or enhanced protein degradation of skeletal muscle cells. The process of skeletal muscle hypertrophy involves changes in multiple signaling pathways, such as IGF-1/PI3K/Akt, myostatin and G protein. On the other hand, activations of the ubiquitin-proteasome system, IGF-1/Akt/FoxO, autophagy-lysosomal pathway, NF-κB, and the glucocorticoid-mediated signaling pathways play important roles in regulating muscle atrophy. These signaling pathways regulate skeletal muscle mass and are modulated by some different conditions. This review briefly summarizes the signaling pathways of skeletal muscle mass control.

[Roles and mechanism of microRNAs in the regulation of skeletal muscle insulin resistance].

Insulin resistance is a common pathophysiological mechanism of obesity and type 2 diabetes mellitus. Skeletal muscle is one of the major target organs of insulin-mediated glucose uptake, metabolism and utilization, and it is the earliest and most important site of insulin resistance. Studies have shown that the impairments of glucose uptake, insulin signaling pathway and mitochondrial biosynthesis are closely related to skeletal muscle insulin resistance. When insulin resistance develops in skeletal muscle, multiple microRNAs (miRNAs) are up-regulated (miR-106b, miR-23a, miR-761, miR-135a, Let-7 and miR-29a) or down-regulated (miR-133a, miR-149 and miR-1). They participate in the regulation of skeletal muscle glucose uptake, insulin signaling pathway and mitochondrial biogenesis, and thus play important roles in the occurrence and development of skeletal muscle insulin resistance. Therefore, these miRNAs may serve as potential targets for the treatment of skeletal muscle insulin resistance or diabetes.

Resolving phylogenetic relationships of Delphacini and Tropidocephalini (Hemiptera: Delphacidae: Delphacinae) as inferred from four genetic loci.

This paper explores the phylogeny of the delphacid subfamily Delphacinae based on nuclear ribosomal and mitochondrial DNA sequences of four genetic loci (16S rDNA, 28S rDNA, Cytochrome oxidase I and Cytochrome b). Maximum likelihood and Bayesian analyses yield robust phylogenetic trees. The topologies support the monophyly of Delphacinae and its basal split into three tribes, and provisionally support subdividing Delphacini into three clades, including a more broadly defined Numatina. The tribe Tropidocephalini is divided into two clades. In addition, Paranectopia is transferred from Tropidocephalini to Delphacini and Harmalia syn. nov. is regarded as a junior synonym of Opiconsiva. The genera Bambusiphaga, Megadelphax and Muirodelphax are found to be paraphyletic. The estimated time to the most recent common ancestor of Delphacinae is roughly at 90 million years ago in Late Cretaceous.

[Fat deposition in skeletal muscle and its regulation].

Under normal condition, there are a few lipid droplets in skeletal muscle. But in skeletal muscle acute injury, muscular dystrophy, muscle atrophy, obesity, diabetes and other pathological conditions, the fat deposition in skeletal muscle increases, which implicate that the fat deposition may play an important role in the pathogenesis of these diseases. However, the mechanisms of development and regulation of fat deposition in skeletal muscle are not clear. Clarifying the key signaling pathways and regulatory factors that affect fat deposition in skeletal muscle, and exploring new ways to improve the fat deposition in skeletal muscle will not only help to deepen our understanding of the pathogenesis of these diseases, but also provide new ideas for the treatment of these diseases. This paper reviews the research progresses and main mechanisms of fat deposition in skeletal muscle.

[The Role of Hepatocyte Growth Factor in Keletal Muscle Regeneration]

Skeletal muscle repair after injury includes three stages which are inflammation, repair and tissue remodeling. The ability of Skeletal muscle to regenerate in response to injury is solely dependen on the activation, proliferation and differentiation of satellite cells. After skeletal muscle injury, hepatocyte growth factor (HGF) can regular muscle satellite cell function in the manner of autocrine, paracrine or endocrine, thus affecting the regeneration of damaged skeletal muscle. Studies about the mechanism have shown that HGF may bind to its receptor, e-met, to start the relevant signaling pathways involved in skeletal muscle satellite cell activation, proliferation, differentiation and migration, which affects skeletal muscle regeneration process.

[Time-Resolved XEOL Experiment System on BL14W1 at SSRF].

A novel time-resolved X-ray excited optical luminescence (TRXEOL) experiment system was developed for X ray absorption fine structure spectroscopy(XAFS) beamline at Shanghai Synchrotron Radiation Facility (SSRF). The TRXEOL system is composed of three parts: timing system, spectrometer system and nuclear instrument module (NIM) system. These three systems were integrated to measure and record the optical luminescence from the sample excited by the synchrotron X-ray pulses, according to the time-correlated single photon counting methodology. It's the first time in the domestic synchrotron radiation facilities to achieve TRXEOL experiment using the synchrotron X-ray pulses and the time structure of the storage ring. In this work, a SSRF-self-developed timing system was used, which is based on the Field programmable Gate Array and the high-speed serial communication technology. The timing system can provide trigger pulse synchronized with the X-ray pulse. The timing jitter is about 6 ps, and the timing delay resolution is 5 ps. The NIM system is the core of the TRXEOL experiment system, it has three main modules: the Constant Fraction Discriminator (CFD), the Time to Amplitude Converter (TAC) and the Multi-Channel Analyzer (MCA). During one excitation circle, the spectrometer and the Photomultiplier Tube detector translate the induced luminescence of the sample excited by a single X-ray pulse into electrical pulse. The CFD module eliminates the timing walk larger than 50 ps induced by the amplitude of the electrical pulse. The TAC module calculates the time interval between the timing trigger pulse and the luminescence electrical pulse, and converts the interval into proportional amplitude of voltage. After plenty of circles, the MCA module gets the luminescence decay curve by recording and analyzing the voltage signals. And the data acquisition system gets the TRXEOL spectra by scanning the spectrometer and acquiring the frequency of the voltage pulses from the TAC module. The TRXEOL experiment system helps researchers measure optical decay curves and spectra of the sample in different time windows. Many luminescence behaviors would be explained more deeply, together with the aid of the optical XAFS to get the electron structure of the sample. A sample of ZnO nanowire was studied using the TRXEOL system. The ordinary XEOL spectrum obtained could distinguish the 390 nm wavelength and the 500 nm wavelength luminescence center. The decay curve at 0 nm wavelength could clearly show the fast luminescence process and the slow luminescence process. The full width at half maximum of the fast luminescence decay curve was about 0.5 ns, showing that the minimum time resolution of the TRXEOL system is less than 1 ns. The TRXEOL spectra obtained could respectively get the luminescence information within different time windows. It was demo nstrated that the TRXEOL system is not only feasible and reliable, but also supply XAFS beamline with the technical preparatio ns of time resolved techniques.

Antioxidant activities of cold-nature Tibetan herbs are signifcantly greater than hot-nature ones and are associated with their levels of total phenolic components.

Tibetan medicinal plants have been used for more than 2 000 years. In order to find their differences in antioxidant activity, total phenolics and total flavonoids between "hot-nature" and "cold-nature" herbs, we investigated the antioxidant activities of 40 Tibetan herbs from Qinghai plateau, with 20 herbs in cold-nature and 20 herbs in hot-nature. Antioxidant capacities were evaluated by the following methods: scavenging ABTS•(+) (2, 2'azinobis-(3-ethylbenz-thiazoline-6-sulfonic acid), scavenging O2•(-), and Ferric-reducing antioxidant power (FRAP). The effects on inhibition of mitochondrion lipid peroxidation were determined by measuring the formation of TBARS (Thiobarbituric acid reactive substrates). Total phenolics and flavonoids were estimated by Folin-Ciocalteu and NaNO2-Al(NO3)3-NaOH colorimetric methods. Interestingly, the cold-nature herbs displayed higher antioxidant activities than the hot-nature ones, corresponding to nearly three-fold higher total phenolic contents in the cold-nature herbs. Moreover, the antioxidant activities correlated linearly with the levels of total phenolics for both cold-nature and hot-nature herbs, but only with the levels of total flavonoids for the hot-nature herbs. The results suggested that the phenolic compounds, but not the flavonoids, play the major role in antioxidant capacities of the cold-nature herbs. These findings could shed new lights on the study the theory of Tibetan medicine.

Synthesis and biological evaluation of hydroxycinnamic acid hydrazide derivatives as inducer of caspase-3.

In order to generate compounds with superior antitumor activity and reduced toxicity, twelve new hydroxycinnamic acid hydrazide derivatives were synthesized and evaluated for their antiproliferative activities against two cancer cell lines (H1299 lung carcinoma cells and MCF-7 breast cancer cells), and compared to two normal counterparts (NL-20 lung epithelial cells and H184B5F5/M10 breast cells) by MTT method. The results demonstrated that some of these compounds possessed good antiproliferative activity against the two cancer cell lines. Among them, compound 2c was active against the growth of H1299 lung carcinoma cells with IC50 values of 1.50 µM, which was more active than the positive topotecan (IC50 = 4.18 µM). Simultaneously, it showed lower cytotoxic effects on normal NL-20 lung epithelial cells (IC50 > 10 µM). Mechanism studies indicated that it induced cell cycle arrest at G2/M phase followed by activation of caspase-3, and consequently caused the cell death. Further studies on the structure optimization are ongoing.

Lauriana Ren & Qin, a new genus of the tribe Tropidocephalini (Hemiptera: Fulgoromorpha: Delphacidae) from China.

A new delphacid genus and species, Lauriana senticosa Ren & Qin, gen. et sp. nov. (Hemiptera: Delphacidae: Tropidocephalini) is described from Sichuan, China. Habitus photos and illustrations of male genitalia of the new species are given, and the differences between the new genus and its closely related genera are discussed.

Nomenclatural changes in the tribe Empoascini of the subfamily Typhlocybinae (Hemiptera, Cicadellidae).

One genus and species are synonymized in the tribe Empoascini of the subfamily Typhlocybinae. Bhatasca Dworakowska, 1995is a junior synonym of Alebrasca Hayashi & Okada, 1994, Bhatasca rectangulata Qin & Zhang, 2011is a junior synonym of Alebrasca actinidiae Hayashi & Okada, 1994. Furthermore, Bhatasca expansa is (necessarily) transferred to the genus Alebrasca.

Antioxidant-based lead discovery for cancer chemoprevention: the case of resveratrol.

Resveratrol is a well-known natural antioxidant and cancer chemopreventive agent that has attracted much interest in the past decade. Resveratrol-directed compounds were synthesized, and their antioxidant effects against reactive oxygen species (ROS)-induced DNA damage, their prooxidant effects on DNA damage in the presence cupric ions, and their cytotoxic and apoptosis-inducing effects on human promyelocytic leukemia (HL-60) cells were investigated in vitro. It was found that the compounds bearing o-diphenoxyl groups exhibited remarkably higher activities in inhibiting ROS-induced DNA damage, accelerating DNA damage in the presence cupric ions, and inducing apoptosis of HL-60 cells compared with the ones bearing no such groups. The detail mechanism of the structure-activity relationship was also studied by the oxidative product analysis of resveratrol and its analogues with galvinoxyl radical or cupric ions and UV-visible spectra change in the presence cupric ions. This study reveals a good and interesting correlation between antioxidant and prooxidant activity, as well as cytotoxicity and apoptosis-inducing activity against HL-60 cells, and provides an idea for designing antioxidant-based cancer chemoprevention agents.

Prooxidant activity of hydroxycinnamic acids on DNA damage in the presence of Cu(II) ions: mechanism and structure-activity relationship.

The prooxidant effect of hydroxycinnamic acids (HCAs), i.e., caffeic acid (CaA), chlorogenic acid (ChA), sinapic acid (SA), ferulic acid (FA), 3-hydroxycinnamic acid (3-HCA) and 4-hydroxycinnamic acid (4-HCA) on supercoiled pBR322 plasmid DNA strand breakage and calf thymus DNA damage in the presence of Cu(II) ions has been studied. It was found that the compounds bearing ortho-dihydroxyl group (CaA and ChA) or bearing 4-hydroxy-3-methoxyl group (SA and FA) exhibited remarkably higher activity in the DNA damage than the ones bearing no such functionalities. The good correlation between the DNA damaging activity and the oxidative potential of the compounds indicates that the electron transfer between HCAs and Cu(II) plays a crucial role in the reaction. UV-Visible spectral changes demonstrated that CaA or ChA can chelate with Cu(II) as a bidentate ligand, hence facilitating intramolecular electron transfer between CaA or ChA and Cu(II). The involvement of reactive oxygen species (ROS) and Cu(I) ions in the DNA damage were affirmed by the inhibition of DNA breakage using mannitol, glutathione (GSH), catalase and bathocuproinedisulfonic acid (BCDS). These results may have important implications regarding the proposed mechanism of apoptosis induced by phenol and endogenous metal ions.

Bioactive compounds from Paecilomyces tenuipes regulating the function of the hypothalamo-hypophyseal system axis in chronic unpredictable stress rats.

BACKGROUND: A bioactive compound from Paecilomyces tenuipes (BCPT) has an inhibitory effect on monoamine oxidase A (MAO-A) and monoamine oxidase B (MAO-B) in vitro and in vivo, which indicates BCPT may be a potential antidepressant. In this study we aimed to study the antidepressant effects of BCPT in the chronic unpredictable stress (CUS) model in rats and explore underlying mechanisms in the hypothalamic-pituitary-adrenal (HPA) axis. METHODS: The antidepressant effects of BCPT were studied in the chronic unpredictable stress model in rats. Animals were housed isolated, except the control group. Rats were exposed daily to different random stressors from day 1 to 21. Awarding response was detected by calculating the 24-hour consumption of sucrose water. Cortisol (CORT) and adrenocorticotropic hormone (ATCH) contents in serum and arginine vasopressin (AVP) contents in the pituitary body were detected by radio immunoassays. Total RNA of hippocampus or hypothalamus was extracted and subjected to reverse transcription-polymerase chain reaction (RT-PCR) for the measurement of corticotrophin releasing hormone (CRH) mRNA or mineralocorticoid receptor (MR) mRNA and glucocorticoid receptor (GR) mRNA levels. Statistical analyses were performed using one way analysis of variance (ANOVA) followed by Student-Newman-Keuls (SNK) test. RESULTS: Chronic unpredictable stress resulted in reduction of sensitivity to reward and abnormality in the HPA axis in the animal model. BCPT improved the reward reaction as measured by increasing sucrose consumption, remarkably reduced serum CORT and ACTH levels and the AVP content in the pituitary body in the CUS-treated rats, decreased the expression of CRH mRNA, enhanced the expression of hippocampus MR mRNA, GR mRNA and decreased the ratio of MR/GR. CONCLUSIONS: BCPT has potentially antidepressant-like activity and normalized the HPA axis hyperactivity in a CUS model of depression in rats. This may be an important mechanism of its antidepressant effect.

Self-assembly and cytotoxicity study of waterwheel-like dinuclear metal complexes: the first metal complexes appended with multiple free hydroxamic acid groups.

Two waterwheel-like dinuclear complexes [M(2)(PHA)(4)(H(2)O)(2)] (M = Cu(II) (1), Zn(II) (2); HPHA = phthal-hydroxamic acid) appended with four free hydroxamic acid groups, namely, free hydroxamic acid metal complexes (FHAMCs) have been synthesized and characterized. The crystal structure of complex 1 was determined by single crystal X-ray diffraction, which adopts the paddlewheel motif with four bidentate carboxylate ligands joining two Cu(II) ions. The relative cytotoxicities of compounds 1 and 2 against SMMC-7721 and HO-8910 cell lines are similar and more predominant than HPHA (IC(50): Cu(II)>Zn(II)>>HPHA). The synergic effect of the bound water molecules, multiple free hydroxamic acid groups and dimetal active sites with bridging carboxylate may have significant impacts on their pharmacological activity. As the prototype for a new class of hydroxamic acid derivatives, the self-assembly of FHAMCs presents a promising new strategy in designing multiple hydroxamic acids with remarkable bioactivities.

DNA damage induced by resveratrol and its synthetic analogues in the presence of Cu (II) ions: mechanism and structure-activity relationship.

The prooxidant effect of resveratrol (3,5,4'-trihydroxy-trans-stibene) and its synthetic analogues (ArOH), that is, 3,4,4'-trihydroxy-trans-stibene (3,4,4'-THS), 3,4,5-trihydroxy-trans-stibene (3,4,5-THS), 3,4-dihydroxy-trans-stibene (3,4-DHS), 4,4'-dihydroxy-trans-stibene (4,4'-DHS), 2,4-dihydroxy-trans-stilbene (2,4-DHS), 3,5-dihydroxy-trans-stilbene (3,5-DHS) and 3,5,4'-trimethoxy-trans-stibene (3,5,4'-TMS), on supercoiled pBR322 plasmid DNA strand breakage and calf thymus DNA damage in the presence of Cu (II) ions has been studied. It was found that the compounds bearing ortho-dihydroxyl groups (3,4-DHS, 3,4,4'-THS, and 3,4,5-THS) or bearing 4-hydroxyl groups (2,4-DHS, 4,4'-DHS, and resveratrol) exhibit remarkably higher activity in the DNA damage than the ones bearing no such functionalities. Kinetic analysis by UV-visible spectra demonstrates that the formation of ArOH-Cu (II) complexes, the stabilization of oxidative intermediate derived from ArOH and Cu (II)/Cu (I) redox cycles, might be responsible for the DNA damage. This study also reveals a good correlation between antioxidant and prooxidant activity, as well as cytotoxicity against human leukemia (HL-60 and Jurkat) cell lines. The mechanisms and implications of these observations are discussed.

Proteasome-independent down-regulation of estrogen receptor-alpha (ERalpha) in breast cancer cells treated with 4,4'-dihydroxy-trans-stilbene.

Treatment of cells with estrogens and several pure ERalpha antagonists rapidly induces down-regulation of the alpha-type estrogen receptor (ERalpha) in the nucleus by mechanisms that are sensitive to the proteasome inhibitors, MG132 and clasto-lactacystin-beta-lactone. Hence, it is believed that these ER ligands induce down-regulation of ERalpha by proteasome-dependent mechanisms, which serve to control both the amount of transcriptional activity and the level of ligand-bound ERalpha in cells. In this study, we observed that treatment of cultured MCF-7 and T47D human breast cancer cells with the low affinity ER ligand, 4,4'-dihydroxy-trans-stilbene (4,4'-DHS), inhibited the transcriptional activity of ERalpha and induced slow and gradual decrease in the amount of ERalpha protein (henceforth referred to as down-regulation of ERalpha). The 4,4'-DHS-induced down-regulation of ERalpha in MCF-7 cells involved a mechanism that was insensitive to the two most specific proteasome inhibitors, clasto-lactacystin-beta-lactone and epoxomycin, but sensitive to MG132 at concentrations exceeding that required for maximal inhibition of the proteasome in MCF-7 cells. Therefore, 4,4'-DHS appears to induce down-regulation of ERalpha by a proteasome-independent mechanism. Here, we present data to show that both 4-OH and 4'-OH are critical for the ability of 4,4'-DHS to induce down-regulation of ERalpha and suggest that 4,4'-DHS provides a useful scaffold for development of novel ERalpha antagonists.