Background and Aims The E2F3 transcription factor claims its role in controlling cell cycle progression. As reported earlier, nuclear E2F3 overexpression leads to development of bladder and prostate ...cancer in humans. Accordingly, the present investigation has been designed to assess to what extent E2F3 would be overexpressed in breast cancer. The aim of this study was to emphasize that the levels of E2F3 are increased in breast cancer and highlights the efficacy of siRNA targeted to E2F3. Methods To investigate the expression level of E2F3 and the progression of breast tumors, quantitative real-time PCR analysis was carried out. Western blotting analysis was performed to measure its counterparts, namely, E2F3a and E2F3b. Results In the novel axis of E2F3, a large set of 11 breast cancer cell lines were identified to have the property of overexpression. Furthermore, the small interfering RNA (siRNA) developed against E2F3 significantly blocked the expression of the E2F3 in the selected breast cancer cell lines. Thus, the present findings authenticate the efficiency of siRNA (E2F3) to fight against breast cancer; hence, the siRNA mediated E2F3 gene silencing knockdown the E2F3. Conclusions This in vitro study demonstrates that E2F3 is a newly identified diagnostic and potential therapeutic target in breast cancer. Outcomes of this study affirm that siRNA for E2F3 facilitates the silencing of E2F3 overexpression and fights against breast cancer. Therefore, it plays a vital role as an alternative for diagnosis and clinical outcome for the treatment of breast cancer.
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Modern therapies for malignant breast cancer in clinics are not efficacious and often result in deprived patient compliance owing to squat therapeutic effectiveness and strong ...systemic side effects. In order to overcome this, we combined chemo-photothermal targeted therapy of breast cancer within one novel multifunctional drug delivery system. Folic Acid-functionalized polyethylene glycol coated Zinc Oxide nanosheet (FA-PEG-ZnO NS), was successfully synthesized, characterized and introduced to the drug delivery field for the first time. A doxorubicin (DOX)-loaded FA-PEG-ZnO NS based system (DOX-FA-PEG-ZnO NS) showed stimulative effect of heat, pH responsive and sustained drug release properties. Cytotoxicity experiments confirmed that combined therapy mediated the maximum rate of death in breast cancer cells compared to that of single chemotherapy or photothermal therapy. In vivo toxicity evaluation showed that the DOX-FA-PEG-ZnO NS contains minimum systemic toxicity in the mice model system. The findings of the present study provided an ideal drug delivery system for breast cancer therapy due to the advanced chemo-photothermal synergistic targeted therapy and good drug release properties of DOX-FA-PEG-ZnO NS, which could effectively avoid frequent and invasive dosing and improve patient compliance. Thus, functionalized-ZnO NS could be used as a novel nanomaterial for selective chemo-photothermal therapy.
•ZnONPs were biologically synthesized using palm fruit extract as reducing and stabilizing agent.•The size and shape of bio-synthesized ZnONPs depends on the concentrations of the extract.•In vitro ...studies on ZnONPs induce a significant reduction of Bcl-2 expression on MCF-7 cells.•In vivo evaluation verified the biocompatibility of DOX-ZnONPs, no significant toxicity on blood stream as well.
The green synthesis of zinc oxide nanoparticles (ZnONPs) using Borassus flabellifer fruit extract was characterized by UV–visible spectroscopy, FT-IR, XRD, TEM, Zeta potential and EDS analysis. The UV–visible spectrum showed an absorption peak at 368nm that reflects surface Plasmon resonance (SPR) ZnONPs. TEM photograph showed that the green synthesized ZnONPs were porous in nature and rod like structure with an average size of 55nm. The Zeta potential value of −21.5mV revealed the surface charge of green synthesized ZnONPs. In this study, we examined the synthesized DOX-ZnONPs exhibited a dose-dependent cytotoxicity against MCF-7 and HT-29. The inhibitory concentration (IC50) was found to be 0.125μgmL−1 for MCF-7 and HT-29 cells. An induction of apoptosis was evidenced by nuclear stain Hoechst 33258. In vivo toxicity assessment showed that DOX-ZnONPs have low systemic toxicity in murine model system. The results prove that the DOX-ZnONPs has low toxicity and high therapy efficacy, which provides convincing evidence for the green biosynthesized ZnO as a promising candidate for a drug delivery system.
Acacia nilotica is widely distributed in Asia. In India, it occupies an important place in the indigenous system of medicine against anti-inflammatory, antioxidant, cancers, and/or tumors.
The ...purpose of this study is to investigate the inhibitory effect of Acacia nilotica leaves extract and γ-Sitosterol on cell proliferation, the apoptotic effect and cell cycle arrest in breast and lung cancer cells.
GC–MS and HPLC were used to determine the chemical constituents of this extract and γ-Sitosterol respectively. Human MCF-7 and A549 cell lines were treated with Acacia nilotica extract and γ-Sitosterol. Cell viability was determined by MTT assay. Cell proliferation was determined by BrdU incorporation assay. Apoptosis was detected by cell morphologic observation through AO/EtBr staining, cell cycle analysis, and immunoblot analysis on the expression of protein associated with cell cycle arrest.
Experimental results of bioactive compound analysis indicate that γ-Sitosterol, bioactive ingredients of Acacia nilotica extract. The IC50 value of extract on MCF-7 and A549 cancer cells was 493.3±15.2 and 696.6±11.5μg/ml, respectively. Acacia nilotica extract and γ-Sitosterol were inhibited the cell proliferation by 54.34±1.8 and 42.18±3.9% for MCF-7 and 58.26±1.5 and 44.36±3.05% for A549 cells. The percentage of apoptotic cells observed in the MCF-7 and A549 cell lines were increased to 42.46 and 36.8% of extract; 46.68 and 43.24% for γ-Sitosterol respectively. Flow cytometric analysis results demonstrate that cells were arrested at the G2/M phase and decrease the c-Myc expression.
This study demonstrates in vitro results, which support the ethnomedical use of γ-Sitosterol against cancer. Experimental results of this study suggest that γ-Sitosterol exerts potential anticancer activity through the growth inhibition, cell cycle arrest and the apoptosis on cancer cells.
Experimental evidence from the etiology of cancer studies suggests that a correlation between Bisphenol-A (BPA) exposure and alterations in hematopoiesis leads to blood cancer. In our study zebrafish ...were used to assess the lethality, developmental effect, embryonic apoptosis and changes in transcription factor of hematopoiesis through EGFR/ERK signaling pathways in response to BPA. The in silico interaction of EGFR and BPA was analysed by molecular dynamic simulation. According to our results, BPA induced a significant lethal effect in hatching retardation, reduction in heart rate and teratogenic effects on zebrafish embryos and larvae at three different concentrations 100, 500 and 2500 μg/L. The mortality of adult zebrafish exposed to the acute toxicity of BPA from 5 to 30 mg/L concentrations was determined for 96 h. The peripheral blood cells and vital organs such as kidney, liver and spleen from BPA exposed fish showed predominantly abnormal myeloid blast cells along with severe morphological changes in erythrocytes at sublethal concentration 245 μg/L. The BPA showed the highest binding affinity to zebrafish EGFR with a docking score of −7.5 kcal/mol with an RMSD of 3.0 nm during MD simulation. We found that EGFR/ERK overexpression leads to induce hematopoietic cell proliferation and impaired differentiation, which enhances the myeloid repopulating activity and the accumulation of immature myeloblast cells. BPA also caused a corresponding increase in expression of hematopoietic transcription factor c-MYB and RUNX-1 leading to polychromasia, poikilocytosis, acanthocytes and anisocytosis and promoted myeloblastosis by inhibiting GATA-1 expression. These morphological changes often resulted in the prior condition of acute myeloid leukemia (AML). Comprehensively, our data suggest that BPA can trigger the malignancy of AML cells by alteration of respective hematopoietic transcription factors via EGFR/ERK signaling in the zebrafish model.
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•Bisphenol A showed an acute toxicity towards embryos, larvae and adult zebrafish.•Bisphenol A caused developmental toxicity and cell apoptosis of zebrafish embryos.•BPA exposed fish showed predominantly abnormal myeloid blast cells along with severe morphological changes in erythrocytes.•BPA showed the highest binding affinity to zebrafish EGFR using docking, RMSD and MD simulation.•BPA caused altering the expression of hematopoietic transcription factor GATA-1, c-MYB and RUNX-1.
Hexavalent chromium Cr(VI), a toxic heavy metal, is a serious pollutant of tannery effluent, and its accumulation in soil and water causes severe environmental concerns of increasing public health ...issues. The present study focus on the isolation and identification of chromium-reducing bacteria collected from the tannery industry in Dindigul, Tamil Nadu. Chromium-reducing bacteria Pseudomonas putida were identified by 16S rRNA sequencing followed by BLAST search. The plasmid with Cr(VI) reductase gene was isolated from Pseudomonas putida and transferred to E. coli DH5α for further studies.
The bacterial cultures were kept under controlled conditions for 72 h to observe the growth rates and bacterial resistance to chromium. When strains wild type and transformant E. coli DH5α were grown in chromium supplemented media revealed significant growth, but strains cured type Pseudomonas putida and E. coli DH5α were minimum growth. The Cr(VI) reduction employed by transformant E. coli DH5α and wild Pseudomonas putida was 42.52 ± 1.48% and 44.46 ± 0.55%, respectively. The culture supernatant of the wild Pseudomonas putida and transformant E. coli DH5α showed an increased reduction of Cr(VI) compared to cell extract supernatant and cell debris due to the extracellular activity of chromium reductase has been responsible for Cr(VI) reduction. Besides, the chromium reductase gene was confirmed in the isolated Pseudomonas putida and transformant E. coli DH5α.
Transformant bacteria could employ an alternative method for heavy metal detoxification in contaminated environments like tannery effluent and mining processes.
Controlling biofilms of bacteria is a challenging aspect because of their drug-resistance potentials against a range of antibiotics, demanding the development of active anti-biofilm agents. ...Rutin (R), a natural antioxidant, and benzamide (B), a synthetic antibacterial agent, have several pharmacological and antibacterial abilities. Herein, we developed PEG–PLGA NPs that synergistically carried rutin and benzamide as drug candidates, while displaying therapeutic and anti-biofilm functions. These drug delivery NPs were synthesized by the oil-in-water emulsion (O/W) solvent evaporation technique. The obtained NPs were characterized by UV–vis, FT-IR, SEM, TEM, and DLS measurements. Confocal laser scanning microscopy was employed to evaluate the anti-biofilm capabilities against Staphylococcus aureus and Pseudomonas aeruginosa and further quantified the levels of residual biofilm constituents such as protein and exopolysaccharide (EPS). Drug release experiments showed the controlled release of rutin–benzamide (RB) for several days. Antibacterial analyses showed that the minimum inhibitory concentration (MIC) of NPs was at least two times lower than that of the free drugs. RB–PEG-PLGA NPs revealed that they targeted biofilm-forming bacteria through the disruption of the membrane and biofilm surface and were observed to be nontoxic when tested using human erythrocytes and human cell lines. In vivo evaluations in zebrafish showed that the NPs did not alter the antioxidant functions and histological features of tissues. On the basis of results obtained, it is substantiated that the rutin–benzamide-loaded nanocarrier offers potential anti-biofilm therapy due to its high anti-biofilm activity and biocompatibility.
Animal venoms and toxins are potential bioresources that have been known to mankind as a therapeutic tool for more than a century through folk and traditional medicine. The purified “disintegrin ...protein” (64kDa) from the venom of the Indian cobra snake (Naja naja) exhibited cytotoxic effects of various types of human cancer cell lines such as breast cancer (MCF-7), lung cancer (A549) and liver cancer (HepG2). In vitro cytotoxicity, DNA fragmentation, an apoptotic assay and a cell cycle analysis were performed to evaluate the anticancer activity of disintegrin against the above cell lines. The IC50 value of disintegrin was determined to be 2.5±0.5μg/mL, 3.5±0.5μg/mL, and 3±0.5μg/mL for the MCF-7, A549 and HepG2 cell lines respectively. Moreover, the increased distribution of G0/G1 and S phase led to decreased populations of cells in the G2/M phase of MCF-7, HepG2 and A549 cells.
Hexavalent chromium (Cr(VI)), a toxic heavy metal, is a serious pollutant from tannery effluent, and its accumulation in soil and water causes severe environmental concerns and increasing public ...health issues. The present study focuses on the isolation and identification of chromium-reducing bacteria collected from the tannery industry in Dindigul, Tamil Nadu. Chromium-reducing bacteria Pseudomonas putida were identified by 16S rRNA sequencing followed by BLAST search. The plasmid with Cr(VI) reductase gene was isolated from Ps. putida and transferred to Escherichia coli DH5α for further studies.
The bacterial cultures were kept under controlled conditions for 72 h to observe the growth rates and bacterial resistance to chromium. When strains wild-type and transformant E. coli DH5α were grown in chromium-supplemented media, they revealed significant growth, but strains cured type Ps. putida and E. coli DH5α recorded minimum growth. The Cr(VI) reduction employed by transformant E. coli DH5α and wild Ps. putida was 42.52 ± 1.48% and 44.46 ± 0.55%, respectively. The culture supernatant of the wild Ps. putida and transformant E. coli DH5α showed an increased reduction of Cr(VI) compared with cell extract supernatant and cell debris due to the extracellular activity of chromium reductase being responsible for Cr(VI) reduction. Besides, the chromium reductase gene was confirmed in the isolated Ps. putida and transformant E. coli DH5α.
Transformant bacteria could employ an alternative method for heavy metal detoxification in contaminated environments like tannery effluent and mining processes.
High Cr(VI) concentration resistance and high Cr(VI) reducing the strain's ability make it suitable for bioremediation. These possible horizontal gene transfer events indicated in this study may have enabled transformant E. coli DH5α as a good candidate for reducing the heavy metal pollution.
Cancer nanotheranostic materials are useful in real-time monitoring of drug delivery and therapeutic action against tumor cells as they co-deliver therapeutic and imaging functions. In this study, we ...report the use of natural fluorescent protein (R-phycoerythrin (PE)) in the preparation of novel multifunctional chitosan–silver–phycoerythrin nanocomposites (CS–Ag–PE NCs) and the theranostic potentials of the synthesized nanocomposites (NCs) were evaluated against triple negative breast cancer (MDA-MB-231) cells. Absorption behavior of the synthesized CS–Ag NCs and CS–Ag–PE NCs at different pH was studied by UV-Vis and FT-IR spectroscopy for determining the interaction of silver with chitosan and CS–Ag NCs with PE. Morphological features of CS–Ag NCs and CS–Ag–PE NCs and cellular uptake of CS–Ag–PE NCs were evaluated by TEM. Flow cytometry analyses were performed to determine the conjugation efficiency of CS–Ag-NCs to PE, stability of the CS–Ag–PE NCs and cellular localization of NCs at dose-dependent concentrations. Further, we substantiated the effect of CS–Ag–PE NCs in the activation of ROS mediated caspase-dependent intrinsic apoptosis by analyzing the expression of apoptotic proteins and bcl-2 family genes, respectively. The ionic interaction between PE and CS–Ag NCs resulted in a stable theranostic fluorescent NC complex that enabled the real time probing of delivery, distribution and therapeutic functions of NCs in cancer cells. The theranostic NCs of this study exhibited apoptotic induction potential in cancer cells but low toxicity in normal breast cells.
