Copper and zinc interact significantly in their joint toxicity to Chlamydomonas reinhardtii: Insights from physiological and transcriptomic investigations.

Copper (Cu) and zinc (Zn) often discharge simultaneously from industrial and agricultural sectors and cause stress to aquatic biota. Although microalgae have been extensively investigated for their responses to Cu or Zn exposure, how they cope with the mixtures of two metals, especially at transcriptomic level, remains largely unknown. In this study, Chlamydomonas reinhardtii was exposed to environmentally relevant concentrations of two metals. It was found that Zn promoted the entry of Cu into the algal cells. With the increase of combined toxicity, extracellular polymeric substances (EPS) and cell wall functional groups immobilized significant amounts of Cu and Zn. Furthermore, C. reinhardtii adjusted resistance strategies internally, including starch consumption and synthesis of chlorophyll and lipids. Upon high level of Cu and Zn coexistence, synergistic effects were observed in lipid peroxidation and catalase (CAT) activity. Under 1.05 mg/L Cu + 0.87 mg/L Zn, 256 differentially expressed genes (DEGs) were mainly involved in oxidative phosphorylation, ribosome, nitrogen metabolism; while 4294 DEGs induced by 4.21 mg/L Cu + 3.48 mg/L Zn were mainly related to photosynthesis, citric acid cycle, etc. Together, this study revealed a more comprehensive understanding of mechanisms of Cu/Zn detoxification in C. reinhardtii, emphasizing critical roles of photosynthetic carbon sequestration and energy metabolism in the metal resistance.

Effects of algal-bacterial ratio on the growth and cadmium accumulation of Chlorella salina-Bacillus subtilis consortia.

Algae-bacteria consortia have been proven effective in the removal of metal pollutants, but the effects of algal-bacterial ratio in the metal accumulation and resistance by this symbiotic system have not been systematically investigated. In this study, we set up consortia with various ratios of Chlorella salina-Bacillus subtilis, determined their growth, Cd accumulation, levels of intracellular glutathione (GSH), extracellular polysaccharide, phosphorus (P) in the culture medium, and functional groups of consortia after Cd treatments (0.1, 0.5, 1 mg L-1 ) for 7 days. With the addition of B. subtilis in the C. salina culture, the dry weight and specific growth rate of the consortia significantly increased compared with C. salina alone, reaching 68.33 mg and 0.382 (mg L-1 ) d-1 respectively at the 1:4 algal-bacterial ratio with 1 mg L-1 Cd treatment. Maximum Cd removal (51.66%) was also observed upon the same Cd exposure and algal-bacterial ratio. Cadmium was mostly taken up into cells at 1 mg L-1 Cd whereas its adsorption dominated the accumulation when Cd was 0.1 and 0.5 mg L-1 . The amounts of extracellular polysaccharides, GSH, and P of the symbiotic system were also increased by the bacterial addition. Besides, Fouriertransform infrared (FTIR) spectroscopy analysis showed that functional groups like N-H, O-H, and P-O-C were involved in the Cd complexation. Taken together, a higher bacterial ratio promoted the Cd accumulation and detoxification by the C. salina-B. subtilis consortia through intra- and extracellular processes.

[Differences in the tolerance and accumulation of arsenate in the consortia with various proportions of Chlorella salina and Bacillus subtilis]. / 不同比例盐生小球藻-æž¯è‰èŠ½å­¢æ†èŒå ±ç”Ÿä½“å¯¹ç ·é ¸ç›è€æ€§åŠå¯Œé›†å·®å¼‚.

We constructed consortia of Chlorella salina and Bacillus subtilis with various alga-bacterium ratios (1:0, 1:1, 1:2, 1:3, 1:4). After being treated with arsenate [As(Ⅴ)] for 7 d, we measured the growth, As accumulation, adsorption and absorption, and As speciation transformation of consortia. Results showed that the chlorophyll content, dry weight, and specific growth rate of the symbiont increased significantly with increasing B. subtilis ratio after the As(Ⅴ) treatment, being 1.81 mg·L-1, 125.0 mg, and 0.28 mg·L-1·d-1 under the condition of the alga-bacterium ratio being 1∶4 and As(Ⅴ) being 750 µg·L-1, respectively. The accumulation and absorption of As by the consortia decreased with the bacterial proportion increasing from 1:0 to 1:4. As accumulation changed with the As concentration, with a dominance of absorption under 75-150 µg·L-1 As(Ⅴ) and a dominance of adsorption under 300-750 µg·L-1As(V). There were As(Ⅴ) and As(Ⅲ) in the consortia. When the proportion of bacteria increased, the rate of As(Ⅴ) reduction enhanced (up to 12.6%). Our results indicated that the increases of B. subtilis improved As(Ⅴ) tolerance and reduction, but decreased the As(Ⅴ) accumulation by the symbiont.

Extracellular polymeric substances alter cell surface properties, toxicity, and accumulation of arsenic in Synechocystis PCC6803.

Arsenic (As) contamination of water poses severe threats to human health and thus requires effective remediation methods. In this study, Synechocystis PCC6803, a model cyanobacterium common in aquatic environments, was used to investigate the role of extracellular polymeric substances (EPS) in As toxicity, accumulation, and transformation processes. We monitored the growth of Synechocystis with As exposure, measured the zeta potential and binding sites on the cell surface, and analysed As accumulation and speciation in Synechocystis cells with and without EPS. After EPS removal, the binding sites and zeta potential of the cell surface decreased by 44.43% and 31.9%, respectively. The growth of Synechocystis decreased 49.4% and 43.7% with As(III) and As(V) exposure, and As accumulation in the cells decreased by 12.8-44.5% and 14-42.7%, respectively. As absorption was enhanced in cells with EPS removed. The oxidation of As(III) and reduction of As(V) were significantly greater in cells with intact EPS compared to those with EPS removed. Fourier transform infrared spectroscopy (FTIR) showed that functional groups of EPS and Synechocystis cells, including -NH, -OH, CO, and CC, interacted with As species. Together the results of this work demonstrate that EPS have significant impacts on cell surface properties, thereby affecting As accumulation and transformation in Synechocystis PCC6803. This work provides a basis for using EPS to remedy As pollution in aquatic environments.

Recombinant human insulin-like growth factor-binding protein 3 inhibits tumor growth and targets the Akt pathway in lung and colon cancer models.

OBJECTIVE: Insulin-like growth factor-binding protein 3 (IGFBP-3) can induce antiproliferative and proapoptotic effects in human cancer cells, by IGF-I independent mechanisms. The antitumor efficacy of recombinant human IGFBP-3 (rhIGFBP-3) and its interaction with chemotherapy in lung and colon cancers, in vitro and in vivo was evaluated. The effects of the different treatments on IGF-IR signaling pathways were also examined. DESIGN: Antiproliferative in vitro assay using rhIGFBP-3, as single agent or in combination with carboplatin or irinotecan against the murine Lewis Lung (M-3LL) and LoVo cell lines, respectively was performed. In the M-3LL model in vivo model, mice were treated with rhIGFBP-3 (3 or 10 mg/kg), carboplatin (25 or 50 mg/kg) alone or in combined treatments. In the LoVo xenograft model, mice were treated with rhIGFBP-3 (3, 10 or 30 mg/kg), irinotecan (10 or 20 mg/kg), as monotherapies or in combinations. RESULTS: rhIGFBP-3 elicited a dose-dependent tumor growth inhibition on the M-3LL model and produced a significant tumor growth inhibition at the highest dose tested. However, it failed to improve the antitumor response to carboplatin. In the LoVo colorectal xenograft model, rhIGFBP-3 caused significant single-agent inhibitory effect and enhanced the antitumor activity of irinotecan at their lowest doses tested. Western blot analysis suggests that the observed tumor growth inhibition by rhIGFBP-3 correlates with decreased Akt phosphorylation in both M-3LL and LoVo cell lines in vitro. CONCLUSIONS: Our novel findings provide evidence for in vivo activity of rhIGFBP-3 against lung and colon tumor models and reveal new insight into its interaction with chemotherapeutic drugs. The antitumor effects of rhIGFBP-3 are associated with a downregulation of AKT signaling.

Recombinant human insulin-like growth factor binding protein 3 inhibits growth of human epidermal growth factor receptor-2-overexpressing breast tumors and potentiates herceptin activity in vivo.

Clinical studies indicate that Herceptin (trastuzumab), a recombinant humanized monoclonal antibody directed against the human epidermal growth factor receptor-2 (HER-2) tyrosine kinase growth factor receptor, provides a significant but transient survival advantage to a subset of patients with HER-2-overexpressing metastatic breast cancer when given as a first-line agent. Increased insulin-like growth factor (IGF)-I receptor (IGF-IR) signaling has recently been identified as a potential factor adversely influencing the response to Herceptin. We examined the effect of recombinant human IGF binding protein 3 (rhIGFBP-3), an antagonist of IGF-IR signaling, in Herceptin-resistant breast cells in vitro and in tumors in vivo. Consistent with results obtained using HER-2- or IGF-IR-transfected cells (MCF-7/HER2-18 and SKBR3/IGF-IR, respectively), we found that rhIGFBP-3 significantly reduced IGF-I-induced IGF-IR phosphorylation and displayed a synergistic interaction with Herceptin against cultured HER-2-overexpressing breast cancer cells in vitro. We show, for the first time, the antitumor activity of rhIGFBP-3 against advanced-stage MCF-7/HER2-18-transfected human breast cancer xenografts and its potentiation of Herceptin activity. We also provide evidence that IGF-IR activation counters the early suppressive effect of Herceptin on HER-2 signaling via Akt and p44/p42 mitogen-activated protein kinase (MAPK), and that inhibition of HER-2-overexpressing human breast tumor growth by rhIGFBP-3 is associated with restored down-regulation of Akt and p44/p42 MAPK phosphorylation in vitro and in vivo. These results emphasize the merit of evaluating simultaneous blockade of the HER-2 and IGF-IR pathways using combination therapy with rhIGFBP-3 plus Herceptin in human clinical trials of patients with HER-2-positive breast cancer.

Gene Expression Profiling of 2-(4-Aminophenyl)benzothiazole-resistant MCF-7 Cells Using cDNA Microarrays.

BACKGROUND: CJM126, 2-(4-aminophenyl) benzothiazole, is a potent inhibitor of human-derived breast carcinoma cell lines. Previous studies have shown that CJM126 elicits concentration-dependent, biphasic growth inhibitory effects against a panel of estrogen receptor-positive and receptor-negative human mammary carcinoma cell lines by a mechanism which has not been fully elucidated. MATERIALS AND METHODS: In an effort to understand the mechanism(s) of resistance to CJM126, the present study used cDNA microarrays (Clontech Laboratories, Inc.) representing 1,176 human cancer-related genes to analyze expression profile changes of two CJM126-resistant cell lines, MCF-710nM126 and MCF-710µM126, previously created by exposing MCF-7 cells to 10 nM and 10 µM CJM126, respectively. RESULTS: Expression changes in the CJM126-resistant MCF-7 cell lines were observed in genes involved in a variety of cell signaling pathways. Gene expression changes common to MCF-710nM126 and MCF-710µM126 cells, as compared to sensitive MCF-7wt cells, were the shut-down of transcription factor Oct-2 and the up-regulation of the negative apoptosis regulator MCL-1, the G1-to-S-phase regulator ubiquitin carrier protein and the GTP-binding protein GST1-HS. These findings indicate the association of a CJM126-resistance phenotype with profound gene transcription dysregulation, decreased apoptotic activity and increased proliferation. Specific changes unique to each of the CJM126-resistant cell lines were also observed. Genes involved in the DNA mismatch-repair pathway, such as MSH2, DNA repair protein RAD51 and damage-specific DNA binding protein were down-regulated in MCF-710nM126, while genes involved in the nucleotide-excision repair pathway, such as ERCC1, RFC and PCNA were overexpressed in MCF-710µM126 Conclusion: The differential changes in the DNA-repair pathways between MCF-710nM126 and MCF-710µM126 cell lines indicate that different processes may have been employed to circumvent the growth inhibition produced by exposure to CJM126. This would also suggest that CJM126 may have concentration-dependent mechanisms of growth inhibition.