Heat Shock Protein Gene Expression Research Articles

Erratum: Effects of mercury on reproduction, avoidance, and heat shock protein gene expression of the soil springtail Folsomia candida

Liu YR, Zheng YM, Zhang LM, Luan YX, He JZ. 2010. Effects of mercury on reproduction, avoidance, and heat shock protein gene expression of the soil springtail Folsomia candida. Environ Toxicol Chem 29: 654-659. Table 2 on page 656 should be replaced with the following table:

Heat shock protein gene expression profile may differentiate between rheumatoid arthritis, osteoarthritis, and healthy controls

Objective: Heat shock proteins (Hsps) have been repeatedly implicated in the pathogenesis of rheumatoid arthritis (RA). The aim of this work was to study Hsp mRNA and protein levels to determine whether they can be used to differentiate between RA, osteoarthritis (OA), and healthy controls.Methods: Hsp27, Hsp60, Hsp70, Hsp90α, and HspBP1 mRNA expression was analysed using real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) in 24 RA, 11 OA, and 21 healthy controls. Hsp70 and HspBP1 protein levels were measured in serum using an enzyme-linked immunosorbent assay (ELISA).Results: Hsp gene expression profiles differ significantly between inflammatory (RA) and non-inflammatory (OA) joint diseases, showing significantly increased Hsp27 and Hsp90α mRNA levels in RA synovial tissues. Up-regulated Hsp60 and Hsp90α together with down-regulated Hsp70 and elevated HspBP1/Hsp70 mRNA ratios can be used to differentiate between RA patients and healthy individuals through analysis of peripheral blood samples. Despite increased HspBP1 levels in RA sera, Hsp70 levels and the HspBP1/Hsp70 protein ratio remained identical in the RA patients and healthy individuals, which may contribute to the inhibition of Hsp70 anti-apoptotic activity.Conclusion: Hsp gene expression analysis can be implemented as a new diagnostic approach to facilitate differentiation between RA, OA, and healthy controls.

Use of Enterococcus faecium to improve common sole ( Solea solea) larviculture

The potential of common sole Solea solea (Linnaeus, 1758) for consistent production in recirculating systems is limited by the huge losses usually observed during the early stages of larval development, mainly due to pathogen infections and insufficient nutritional balance. In the hope that use of a suitable probiotic strain may counter these difficulties, in this study we isolated Enterococcus faecium IMC 511 from common sole brood stock and utilized it as probiotic candidate during fish feeding. After a daily administration (twice a day) to sole larvae through live feeds, the intestinal presence of E. faecium IMC 511 was estimated on days 10, 30, and 50 post hatch, and the effects on intestinal microbial load, fish survival, welfare, and growth were evaluated by morphometric and molecular approaches by examining relative body weight, total length, and gene expression of myostatin. In particular, the effect on animal welfare was assessed through analyses of cortisol levels and 70 kDa Heat Shock Protein gene expression. The results showed that while the Vibrio populations of the intestine of S. solea larvae were reduced in a significant way by the probiotic E. faecium IMC 511, survival was not affected. 70 kDa Heat Shock Protein and myostatin gene expression were significantly reduced in probiotic fed larvae with respect to the control group. It is of note that E. faecium IMC 511 seemed to be predominant at day 50 post hatching, probably due to stabilization of the intestinal microbiota, a factor that, together with the lower levels of HSP70, could improve animal well-being and growth. Cortisol levels significantly increased after 10 days of treatment in probiotic fed larvae, while on days 30 and 50 no significant differences were observed among all experimental groups. Considered together, the microbiological data and the molecular and morphometric results suggest that 50 days of probiotic treatment can improve common sole larval growth. Also, it is hoped that these results offer a useful starting point for elucidating the pathways involved in S. solea growth fostered by E. faecium IMC 511 administration.

Effects of anthraquinone extract from Rheum officinale Bail on the physiological responses and HSP70 gene expression of Megalobrama amblycephala under Aeromonas hydrophila infection

We evaluated the effect of dietary supplementation with anthraquinone extract (from Rheum officinale Bail) on the resistance to Aeromonas hydrophila infection in Megalobrama amblycephala. The fish were randomly divided into two groups: a control group (fed a standard diet) and a treatment group (standard diet supplemented with 0.1% anthraquinone extract) and fed for 10 weeks. We then challenged the fish with A. hydrophila and recorded mortality and changes in serum cortisol, lysozyme, alkaline phosphatase (ALP), total protein, aspartate aminotransferase (AST), alanine aminotransferase (ALT), and hepatic catalase (CAT), superoxide dismutase (SOD), malondialdehyde (MDA) and the relative expression of heat shock protein 70 (HSP70) mRNA for a period of 5 d. Supplementation with 0.1% anthraquinone extract significantly increased serum lysozyme activity before infection, serum ALP activity at 24 h after infection, serum total protein concentration 12 h after infection, hepatic CAT activity 12 h after infection, hepatic SOD activity before infection, and the relative expression of hepatic HSP70 mRNA both before infection and 6 h after infection. In addition, the supplemented group had decreased levels of serum cortisol 6 h after infection, serum AST and ALT activities 12 h after infection, and hepatic MDA content 12 h after infection. Mortality was significantly lower in the treatment group (86.67%) than the control (100%). Our results suggest that ingestion of a basal diet supplemented with 0.1% anthraquinone extract from R. officinale Bail can enhance resistance against pathogenic infections in M. amblycephala.

Rickettsia influences thermotolerance in the whitefly Bemisia tabaci B biotype

Abstract The whitefly Bemisia tabaci harbors Portiera aleyrodidarum, an obligatory symbiotic bacterium, as well as several secondary symbionts, including Rickettsia, Hamiltonella, Wolbachia, Arsenophonus, Cardinium and Fritschea, the function of which is unknown. In Israel, Rickettsia is found in both the B and Q of B. tabaci biotypes, and while all other secondary symbionts are located in the bacteriomes, Rickettsia can occupy most of the body cavity of the insect. We tested whether Rickettsia influences the biology of B. tabaci and found that exposing a Rickettsia-containing population to increasing temperatures significantly increases its tolerance to heat shock that reached 40°C, compared to a Rickettsia-free population. This increase in tolerance to heat shock was not associated with specific induction of heat-shock protein gene expression; however, it was associated with reduction in Rickettsia numbers as was assessed by quantitative real-time polymerase chain reaction and fluorescence in situ hybridization analyses. To assess the causes for thermotolerance when Rickettsia is reduced, we tested whether its presence is associated with the induction of genes required for thermotolerance. We found that under normal 25°C rearing temperature, genes associated with response to stress such as cytoskeleton genes are induced in the Rickettsia-containing population. Thus, the presence of Rickettsia in B. tabaci under normal conditions induces the expression of genes required for thermotolerance that under high temperatures indirectly lead to this tolerance.

Ultraviolet B retards growth, induces oxidative stress, and modulates DNA repair-related gene and heat shock protein gene expression in the monogonont rotifer, Brachionus sp.

Ultraviolet B (UV-B) radiation causes direct cellular damage by breakage of DNA strands and oxidative stress induction in aquatic organisms. To understand the effect of UV-B radiation on the rotifer, Brachionus sp., several parameters including 24-h survival rate, population growth rate, and ROS level were measured after exposure to a wide range of UV-B doses. To check the expression of other important inducible genes such as replication protein A ( RPA), DNA-dependent protein kinase ( DNA-PK), Ku70, Ku80, and heat shock proteins ( hsps) after UV-B radiation, we observed dose- and time-dependency at 2 kJ/m 2. We also examined 13 hsp genes for their roles in the UV-B damaged rotifer. Results showed that UV-B remarkably inhibited the population growth of Brachionus sp. The level of intracellular reactive oxygen species (ROS) was high at 2 kJ/m 2, suggesting that 2 kJ/m 2 would already be toxic. This result was supported by other enzymatic activities, such as GSH levels, glutathione peroxidase, glutathione S-transferase, and glutathione reductase. For dose dependency, low doses of UV-B radiation (2, 4, and 6 kJ/m 2) significantly up-regulated the examined genes (e.g. RPA, DNA-PK, Ku70, and Ku80). For the time course study, RPA genes showed immediate up-regulation but returned to basal or lower expression levels compared to the control 3 h after UV-B exposure. The DNA-PK and Ku70/80 genes significantly increased, indicating that they may be involved in repairing processes against a low dose of UV-B exposure (2 kJ/m 2). At the basal level, the hsp90α1 gene showed the highest expression, and followed by hsp10, hsp30, hsp60, and hsc70, and hsp90β in adults (w/o egg). In eggs, the hsp10 gene was expressed the highest, and followed by hsp30, hsp27, hsp90α1, and hsp60 genes. In real-time RT-PCR array on rotifer hsp genes, low doses of UV-B radiation (2 and 4 kJ/m 2) showed up-regulation of several hsp genes but most of the hsp genes showed down-regulation at 8 kJ/m 2 and higher, indicating that significant Hsp-mediated cellular damage already occurred at low doses. For the time course study of four hsp genes ( hsp20, hsp27, hsp70, hsp90α1), they showed a significant correlation for UV-B radiation (2 kJ/m 2). In this paper, we demonstrated that UV-B radiation would affect growth retardation with up- or down-regulation of some important genes in DNA replication, repair process, and chaperoning. This finding provides a better understanding of molecular mechanisms involved in UV-B-mediated cellular damage in the rotifer, Brachionus sp.

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Gene expression profiles of cytosolic heat shock proteins Hsp70 and Hsp90 from symbiotic dinoflagellates in response to thermal stress: possible implications for coral bleaching

Unicellular photosynthetic dinoflagellates of the genus Symbiodinium are the most common endosymbionts of reef-building scleractinian corals, living in a symbiotic partnership known to be highly susceptible to environmental changes such as hyperthermic stress. In this study, we identified members of two major heat shock proteins (HSPs) families, Hsp70 and Hsp90, in Symbiodinium sp. (clade C) with full-length sequences that showed the highest similarity and evolutionary relationship with other known HSPs from dinoflagellate protists. Regulation of HSPs gene expression was examined in samples of the scleractinian coral Acropora millepora subjected to elevated temperatures progressively over 18h (fast) and 120h (gradual thermal stress). Moderate to severe heat stress at 26°C and 29°C (+3°C and +6°C above average sea temperature) resulted in an increase in algal Hsp70 gene expression from 39% to 57%, while extreme heat stress (+9°C) reduced Hsp70 transcript abundance by 60% (after 18h) and 70% (after 120h). Elevated temperatures decreased an Hsp90 expression under both rapid and gradual heat stress scenarios. Comparable Hsp70 and Hsp90 gene expression patterns were observed in Symbiodinium cultures and in hospite, indicating their independent regulation from the host. Differential gene expression profiles observed for Hsp70 and Hsp90 suggests diverse roles of these molecular chaperones during heat stress response. Reduced expression of the Hsp90 gene under heat stress can indicate a reduced role in inhibiting the heat shock transcription factor which may lead to activation of heat-inducible genes and heat acclimation.

ChemInform Abstract: Stresgenin B, an Inhibitor of Heat-Induced Heat Shock Protein Gene Expression, Produced by Streptomyces sp. AS-9.

Stresgenin B was isolated as an inhibitor of heat-induced heat shock protein (HSP) gene expression from a culture broth of Streptomyces sp. AS-9 by silica gel chromatography and HPLC. The molecular formula of the novel compound was determined as C11H13NO5 by high resolution FAB-MS analysis, and the structure was determined by UV, 1H NMR, 13C NMR, HMQC, HMBC, and NOESY spectra. Stresgenin B inhibited heat-induced luciferase reporter-gene expression directed by the human hsp70B promoter in Chinese hamster ovary (CHO) cells at concentrations lower than the concentrations for inhibition of dexamethasone-induced luciferase reporter-gene expression directed by the mouse mammary tumor virus (MMTV)-LTR promoter. The inhibition of heat-induced reporter gene expression was evident even when cells were exposed to stresgenin B only during heat stress treatment. Moreover, the compound inhibited heat-induced syntheses of hsp72/73, hsp90, and hsp110 and thereby suppressed the induction of thermotolerance. Stresgenin B showed moderate cytotoxic activities against several neoplastic cell lines and also showed antibacterial activities against Micrococcus luteus, Bacillus subtilis and Staphylococcus aureus strains.

Heat Shock Protein translocation induced by membrane fluidization increases tumor-cell sensitivity to chemotherapeutic drugs

Treatment of chronic lymphocytic leukemia (CLL) remains a challenge due to the frequency of drug resistance amongst patients. Improving the delivery of chemotherapeutic agents while reducing the expression of anti-apoptotic Heat Shock Proteins (HSPs) within the cancer cells may facilitate in overcoming this drug resistance. We demonstrate for the first time that sub-lethal doses of chemotherapeutic agents can be combined with membrane fluidizing treatments to produce a significant increase in drug efficacy and apoptosis in vitro. We show that fluidizers result in a transient decrease in intracellular HSPs, resulting in increased tumor-cell sensitivity and a membrane-associated induction of HSP gene expression.

Dietary supplementation with l-arginine or N-carbamylglutamate enhances intestinal growth and heat shock protein-70 expression in weanling pigs fed a corn- and soybean meal-based diet

This study determined effects of dietary supplementation with L-arginine (Arg) or N-carbamylglutamate (NCG) on intestinal health and growth in early-weaned pigs. Eighty-four Landrace x Yorkshire pigs (average body weight of 5.56+/-0.07 kg; weaned at 21 days of age) were fed for 7 days one of the three isonitrogenous diets: (1) a corn- and soybean meal-based diet (CSM), (2) CSM+0.08% NCG (0.08%), and (3) CSM+0.6% Arg. There were four pens of pigs per diet (7 pigs/pen). At the end of a 7-day feeding period, six piglets were randomly selected from each treatment for tissue collections. Compared with the control group, Arg or NCG supplementation increased (P<0.05): (1) Arg concentrations in plasma, (2) small-intestinal growth, (3) villus height in duodenum, jejunum and ileum, (4) crypt depth in jejunum and ileum, (5) goblet cell counts in intestinal mucosae, and (6) whole-body weight gain in pigs. Real-time polymerase chain reaction and western blotting analyses revealed that both mRNA and protein levels for heat shock protein-70 (HSP70) were higher (P<0.05) in the intestinal mucosae of Arg- or NCG-supplemented pigs than in the control group. Furthermore, the incidence of diarrhea in the NCG group was 18% lower (P<0.01) than that in the control group. Collectively, these results indicate that dietary supplementation with 0.6% Arg or 0.08% NCG enhances intestinal HSP70 gene expression, intestinal growth and integrity, and the availability of dietary nutrients for whole-body weight gain in postweaning pigs fed a CSM-based diet. Thus, Arg or NCG is a functional ingredient in the weaning diet to improve nutrition, health, and growth performance of these neonates.

Effects of mercury on reproduction, avoidance, and heat shock protein gene expression of the soil springtail Folsomia candida

Based on the Cambisols of Beijing (used as agricultural soils), toxicity tests were conducted to investigate the effects of mercury (Hg) on reproduction and avoidance of Folsomia candida (Hexapoda: Collembola), as well as the transcriptional responses of the hsp70 gene, under different Hg concentrations and at different exposure times. Results showed that the hsp70 gene of the springtail was the most sensitive parameter to soil Hg stress, with a half-maximal effective concentration (EC50) of 0.42 mg/kg. The EC50 values based on reproduction and avoidance tests were 9.29 and 3.88 mg/kg, respectively. The expression level of the hsp70 gene was significantly up-regulated when soil Hg concentration was over 0.25 mg/kg (lowest-observed-effect concentration [LOEC]). In addition, responses of this gene expression were strongly induced after 48 h exposure under 1 mg/kg soil Hg, which probably was due to the fast and sensitive response of the gene transcription to Hg stress. Thus, the results suggested that the responses of the hsp70 gene and individual-level effects (reproduction and avoidance) could be integrated to provide helpful information for environmental monitoring and assessment of contaminated soils.

Effect of dietary probiotics on clownfish: a molecular approach to define how lactic acid bacteria modulate development in a marine fish

We set out to determine whether probiotic addition would improve larval development in the false percula clownfish Amphiprion ocellaris and to determine what molecular responses could be observed in the larvae following probiotic exposure. We supplied the probiotic bacterial strain Lactobacillus rhamnosus IMC 501 to clownfish larvae from the first day posthatch simultaneously by live prey and with addition to rearing water (group 2) and exclusively by live prey (group 3). We observed twofold higher body weight in both clownfish larvae and juveniles when probiotics were supplied via live prey and added to the rearing water. In addition, development was accelerated with metamorphosis occurring 3 days earlier in fingerlings treated with probiotic. Alteration in molecular biomarkers supported the faster growth observation. There was significantly increased gene expression of factors involved in growth and development (insulin-like growth factors I and II, myostatin, peroxisome proliferator-activated receptors alpha and beta, vitamin D receptor alpha, and retinoic acid receptor gamma) when probiotics were delivered via live prey and added to the rearing water. Moreover, probiotic treatment lessened the severity of the general stress response as exhibited by lower levels of glucocorticoid receptor and 70-kDa heat shock protein gene expression. Furthermore, an improvement of skeletal head development was observed, with a 10-20% reduction in deformities for juveniles treated with probiotic. All data suggest a potent effect on development resulting from the administration of lactic acid bacteria to larval clownfish, and this study provides a preliminary molecular entry path into the investigation of mechanisms responsible for probiotic enhancement in fish development.

Small heat‐shock proteins and leaf cooling capacity account for the unusual heat tolerance of the central spike leaves inAgave tequilanavar. Weber

Agaves are perennial crassulacean acid metabolism (CAM) plants distributed in tropical and subtropical arid environments, features that are attractive for studying the heat-shock response. In agaves, the stress response can be analysed easily during leaf development, as they form a spirally shaped rosette, having the meristem surrounded by folded leaves in the centre (spike) and the unfolded and more mature leaves in the periphery. Here, we report that the spike of Agave tequilana is the most thermotolerant part of the rosette withstanding shocks of up to 55 degrees C. This finding was inconsistent with the patterns of heat-shock protein (Hsp) gene expression, as maximal accumulation of Hsp transcripts was at 44 degrees C in all sectors (spike, inner, middle and outer). However, levels of small HSP (sHSP)-CI and sHSP-CII proteins were conspicuously higher in spike leaves at all temperatures correlating with their thermotolerance. In addition, spike leaves showed a higher stomatal density and abated more efficiently their temperature several degrees below that of air. We propose that the greater capacity for leaf cooling during the day in response to heat stress, and the elevated levels of sHSPs, constitute part of a set of strategies that protect the SAM and folded leaves of A. tequilana from high temperatures.

Heat shock protein patterns in the bovine ovary and relation with cystic ovarian disease

The present study was performed to determine how the development of cystic ovarian disease (COD) affecting the ovarian expression of heat shock proteins (HSP) in cows were expressing extrous cycles. HSP27, HSP60, HSP70 and HSP90 were evaluated in different ovarian components by Western blot and semiquantitative immunohistochemical analysis. Greater expression of the HSP27 gene was detected in the granulosa and theca cells of primary, secondary, tertiary and cystic follicles, with decreasing amount in atretic follicles. HSP60, HSP70 and HSP90 showed a similar pattern of immunostaining, with moderate gene expression in primary and secondary follicles, increased expression in tertiary and atretic follicles with the greatest gene expression in cystic follicles. HSP were also localized in the corpus luteum, corpus albicans, interstitial tissue and tunica albuginea. The relative amount of protein in the follicular wall of small and large healthy follicles and cystic follicles as analysed by Western immunoblot was consistent with the immunohistochemical data. We speculate that altered expression of HSP genes decreases apoptosis in the follicular wall and leads to the delayed regression of cystic follicles. This study supports earlier observations suggesting that aberrant HSP gene expression, observed in cells of the cystic follicles, is probably associated with the intra-ovarian component of COD pathogenesis.

A field-study of inducible molecular defenses, ultraviolet radiation, and melanomagenesis in natural Xiphophorus hybrids

Ultraviolet radiation—the primary natural pollutant affecting melanomagenesis—may represent a widespread ecological stressor for many fishes, and yet the relationship between UV-exposure and stress has not been investigated in natural fish populations. Recent lab-based studies have sought to characterize the relationship between tumorigenesis and the induction of molecular defenses, such as heat shock proteins. Here we show that ultraviolet radiation and heat shock protein gene expression explain a significant amount of the variation in hyper-melanization—the phenotypic precursor to melanoma—in wild hybrids of Xiphophorus, laboratory models in cancer research. Our results suggest exposure to UV radiation causes stress which induces molecular defense mechanisms, which in turn may facilitate tumorigenesis in natural fish populations. Studies of laboratory-based model organisms in natural settings, like this one, may provide important insights into ecological and evolutionary relationships obscured in controlled laboratory environments. We hope that ours is only the first of many studies to investigate the such relationships between environmental stress, stress-induced molecular defenses, and cancer in fishes.

Core temperature correlates with expression of selected stress and immunomodulatory genes in febrile patients with sepsis and noninfectious SIRS

Environmental hyperthermia and exercise produce extensive changes in gene expression in human blood cells, but it is unknown whether this also happens during febrile-range hyperthermia. We tested the hypothesis that heat shock protein (HSP) and immunomodulatory stress gene expression correlate with fever in intensive care unit patients. Whole blood messenger RNA was obtained over consecutive days from 100 hospitalized patients suffering from sepsis or noninfectious systemic inflammatory response syndrome (SIRS) as defined by conventional criteria. The most abnormal body temperature in the preceding 24 h was recorded for each sample. Expression analysis was performed using the Affymetrix U133 chip. ANCOVA followed by correlation analysis was performed on a subset of 278 prospectively identified sequences of interest. Temperature affected expression of 60 sequences, either independently or as a function of clinical diagnosis. Forty-eight of these (representing 38 genes) were affected by temperature only, including several HSPs, transcription factors heat shock factor (HSF)-1 and HSF-4, cellular adhesion molecules such as ICAM1/CD54 and JAM3, toll receptors TLR-6 and TLR-7, ribosomal proteins, and a number of molecules involved in inflammatory pathways. Twelve sequences demonstrated temperature-dependent responses that differed significantly between patients with sepsis and noninfectious SIRS: CXCL-13; heat shock proteins DNAJB12 and DNAJC4; the F11 receptor; folate hydrolase 1; HSF-2; HSP 70 proteins HSPA1A, HSPA1B, and HSPA1L; interleukin 8; lipopolysaccharide binding protein; and prostaglandin E synthase. Febrile-range temperatures achieved during sepsis and noninfectious SIRS correlate with detectable changes in stress gene expression in vivo, suggesting that fever can activate HSP gene expression and modify innate immune responses. For some genes, it appears that clinical condition can alter temperature-sensitive gene expression. Collectively, these data underscore the potential importance of body temperature in shaping the immune response to infection and injury.

P-glycoprotein (multi-xenobiotic resistance) and heat shock protein gene expression in the reef coral Montastraea franksi in response to environmental toxicants

The deleterious impacts of marine pollutants on reef corals and their symbiotic algae are an important element of global coral reef decline. In the current study we examined the impacts of toxicants on the reef coral Montastraea franksi by analysing the expression of three stress-related genes belonging to the coral host, using Taqman real-time quantitative reverse transcription-PCR. Gene expression profiles of P-glycoprotein (or multi-xenobiotic resistance protein) (P-gp); heat shock protein 70 (Hsp70) and heat shock protein 90 (Hsp90) were examined following 4 and 8 h exposures to the heavy metal copper (3, 10, 30 and 100 μg L −1) or the third generation oil dispersant Corexit™9527 (1, 5, 10 and 50 ppm). Additionally, the expression of P-gp was examined following exposure to 0.5 and 5 μM concentrations of the chemotherapeutic drug vinblastine, a classic substrate of P-gp. The expression of P-gp increased significantly in corals treated with vinblastine and also increased following exposure to copper and Corexit™9527. Hsp70, and to a lesser extent Hsp90 expression increased following exposure to copper and Corexit™9527 indicating a general cellular stress response. Densities of symbiotic algae in the tissues of the corals did not change significantly during the experiments, nor was any loss or paling of coral tissues observed. These findings provide important insight into how corals defend themselves against pollution and complement ongoing initiatives developing molecular biomarkers of stress in reef-building corals.

Heat shock protein 70 silencing enhances apoptosis inducing factor-mediated cell death in hepatocellular carcinoma HepG2 cells

Heat-shock protein 70 (HSP70) abundantly expressed in human tumors and protects cells from a wide range of apoptotic stimuli. To study the role of HSP70 in human hepatocarcinomas, we stably silenced HSP70 gene expression in HepG2 cells by transfecting the cells with HSP70-silencing vector (HSP70 shRNA). HSP70 silencing resulted in reduced capacities for cell proliferation in vitro and colony formation in soft agar. In addition, HSP70 silencing enhanced chemotherapy drug-induced cell death and tumor growth in a mouse xenograft model. Most importantly, we extended our previous observation that HSP70 modulates mitochondrial protein Apoptosis-inducing factor (AIF)-induced cell death. In this study, we found that HSP70 silencing dramatically increased AIFΔ1-120 mutant (an active form of AIF after deletion of the mitochondria localization domain)-induced apoptosis. In HSP70 silenced cells, AIFΔ1-120 mutant predominantly localized in nuclear compartment compared to the control cells, confirming that HSP70 protects from apoptosis by sequestering AIF in the cytoplasm. Taken together, our data suggest that HSP70 ablation in combination with delivery of AIFΔ1-120 mutant or other chemotherapy drugs represents a potent anti-cancer therapy for hepatocarcinomas.

Triggers of the HSP70 stress response: environmental responses and laboratory manipulation in an Antarctic marine invertebrate (Nacella concinna)

The Antarctic limpet, Nacella concinna, exhibits the classical heat shock response, with up-regulation of duplicated forms of the inducible heat shock protein 70 (HSP70) gene in response to experimental manipulation of seawater temperatures. However, this response only occurs in the laboratory at temperatures well in excess of any experienced in the field. Subsequent environmental sampling of inter-tidal animals also showed up-regulation of these genes, but at temperature thresholds much lower than those required to elicit a response in the laboratory. It was hypothesised that this was a reflection of the complexity of the stresses encountered in the inter-tidal region. Here, we describe a further series of experiments comprising both laboratory manipulation and environmental sampling of N. concinna. We investigate the expression of HSP70 gene family members (HSP70A, HSP70B, GRP78 and HSC70) in response to a further suite of environmental stressors: seasonal and experimental cold, freshwater, desiccation, chronic heat and periodic emersion. Lowered temperatures (-1.9 degrees C and -1.6 degrees C), generally produced a down-regulation of all HSP70 family members, with some up-regulation of HSC70 when emerging from the winter period and increasing sea temperatures. There was no significant response to freshwater immersion. In response to acute and chronic heat treatments plus simulated tidal cycles, the data showed a clear pattern. HSP70A showed a strong but very short-term response to heat whilst the duplicated HSP70B also showed heat to be a trigger, but had a more sustained response to complex stresses. GRP78 expression indicates that it was acting as a generalised stress response under the experimental conditions described here. HSC70 was the major chaperone invoked in response to long-term stresses of varying types. These results provide intriguing clues not only to the complexity of HSP70 gene expression in response to environmental change but also insights into the stress response of a non-model species.