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Berberis vulgaris (L.) has remarkable ethnopharmacological properties and is widely used in traditional medicine. The present study investigated B. vulgaris stem bark (Berberidis cortex) by extraction with 50% ethanol. The main secondary metabolites were quantified, resulting in a polyphenols content of 17.6780 ± 3.9320 mg Eq tannic acid/100 g extract, phenolic acids amount of 3.3886 ± 0.3481 mg Eq chlorogenic acid/100 g extract and 78.95 µg/g berberine. The dried hydro-ethanolic extract (BVE) was thoroughly analyzed using Ultra-High-Performance Liquid Chromatography coupled with High-Resolution Mass Spectrometry (UHPLC–HRMS/MS) and HPLC, and 40 bioactive phenolic constituents were identified. Then, the antioxidant potential of BVE was evaluated using three methods. Our results could explain the protective effects of Berberidis cortex EC50FRAP = 0.1398 mg/mL, IC50ABTS = 0.0442 mg/mL, IC50DPPH = 0.2610 mg/mL compared to ascorbic acid (IC50 = 0.0165 mg/mL). Next, the acute toxicity and teratogenicity of BVE and berberine—berberine sulfate hydrate (BS)—investigated on Daphnia sp. revealed significant BS toxicity after 24 h, while BVE revealed considerable toxicity after 48 h and induced embryonic developmental delays. Finally, the anticancer effects of BVE and BS were evaluated in different tumor cell lines after 24 and 48 h of treatments. The MTS assay evidenced dose- and time-dependent antiproliferative activity, which was higher for BS than BVE. The strongest diminution of tumor cell viability was recorded in the breast (MDA-MB-231), colon (LoVo) cancer, and OSCC (PE/CA-PJ49) cell lines after 48 h of exposure (IC50 < 100 µg/mL). However, no cytotoxicity was reported in the normal epithelial cells (HUVEC) and hepatocellular carcinoma (HT-29) cell lines. Extensive data analysis supports our results, showing a significant correlation between the BVE concentration, phenolic compounds content, antioxidant activity, exposure time, and the viability rate of various normal cells and cancer cell lines.

Capsicum annuum (L.) is one of the essential spices most frequently used in our daily routine and has remarkable ethnobotanical and pharmacological properties. Its fruits are rich in vitamins, minerals, carotenoids, and numerous other phenolic metabolites with a well-known antioxidant activity. Regular consumption of chili fruits may have a positive influence on human health. Therefore, we investigated a commercially available chili fruit powder in the present study, extracting it with 50% ethanol. The dried hydro-ethanolic extract (CAE) was thoroughly analyzed using ultra-high-performance liquid chromatography coupled with high-resolution mass spectrometry (UHPLC–HRMS/MS), and 79 bioactive phenolic constituents were identified. Then, we quantified the main phenolic compounds and found a polyphenol content of 4.725 ± 1.361 mg Eq tannic acid/100 g extract and a flavonoid amount of 1.154 ± 0.044 mg Eq rutin/100 g extract. Phenolic secondary metabolites are known for their dual redox behavior as antioxidants/pro-oxidants, underlying their numerous benefits in health and disease. Thus, the antioxidant potential of CAE was evaluated using three methods; our results could explain the protective effects of chili fruits: IC50DPPH = 1.669 mg/mL, IC50ABTS = 0.200 mg/mL, and EC50FRAP = 0.561 mg/mL. The pro-oxidant potential of phenolic compounds could be a basis for CAE cytotoxicity, investigated in vitro on tumor cell lines and in vivo on Daphnia sp. Results demonstrated the dose- and time-dependent CAE’s cytotoxic activity; the highest antiproliferative activity was recorded on colon (LoVo) and breast (MDA-MB-231) cancer cell lines after 48 h of exposure (IC50 values < 200 µg/mL). In vivo testing on Daphnia sp. reported a potent CAE cytotoxicity after 48 h and embryonic developmental delays. Extensive data analyses support our results, showing a significant correlation between the CAE’s concentration, phenolic compound content, antioxidant activity, exposure time, and the viability rate of different tested cell lines.

To overcome chondrosarcoma's (CHS) high chemo- and radioresistance, we used polyethylene glycol-encapsulated iron oxide nanoparticles (IONPs) for the controlled delivery of the chemotherapeutic doxorubicin (IONPDOX) to amplify the cytotoxicity of proton radiation therapy. Human 2D CHS SW1353 cells were treated with protons (linear energy transfer (LET): 1.6 and 12.6 keV/µm) with and without IONPDOX. Cell survival was assayed using a clonogenic test, and genotoxicity was tested through the formation of micronuclei (MN) and γH2AX foci, respectively. Morphology together with spectral fingerprints of nuclei were measured using enhanced dark-field microscopy (EDFM) assembled with a hyperspectral imaging (HI) module and an axial scanning fluorescence module, as well as scanning electron microscopy (SEM) coupled with energy-dispersive X-Ray spectroscopy (EDX). Cell survival was also determined in 3D SW3153 spheroids following treatment with low-LET protons with/without the IONPDOX compound. IONPDOX increased radiosensitivity following proton irradiation at both LETs in correlation with DNA damage expressed as MN or γH2AX. The IONPDOX–low-LET proton combination caused a more lethal effect compared to IONPDOX–high-LET protons. CHS cell biological alterations were reflected by the modifications in the hyperspectral images and spectral profiles, emphasizing new possible spectroscopic markers of cancer therapy effects. Our findings show that the proposed treatment combination has the potential to improve the management of CHS. [ABSTRACT FROM AUTHOR]

Cisplatin is one of the most effective anticancer agents used to treat colon cancer, which is the third malignancy between the most common human cancers in the world, but the resistance developed represents an obstacle against the full success of chemotherapy. An emerging interest appeared in finding other metallic compounds, such as ruthenium(III) complexes, for chemotherapeutic application in cancer. Our study focused on the anticancer activity of several ruthenium (Ru) complexes with quinolone antibiotics in colon tumor cell cultures. Real-time cell analysis and drug-mediated cytotoxicity tests monitored the inhibitory effects in the drug-treated LoVo colon cancer cells. Flow cytometry assays were performed to evaluate cell cycle phases distribution and apoptotic events. The obtained results showed dose-dependent increased levels of cell lysis and induction of apoptosis in LoVo cancer cells treated with the Ru(III) complexes. In addition, data showed a major decrease in cell proliferation, since the percentages of cells distributed in the S cell cycle phase diminished, and a G0/G1 cell arrest was observed. Therefore, our results strongly suggest that the newly synthesized Ru(III) complexes might play an important role in future chemotherapeutic approaches, since their activity is based on diminishing cell proliferation, induction of apoptosis, and modulation of cell cycle phases.

New pyrrolo[1,2-b]pyridazines were synthesized by 3 + 2 cycloaddition reaction between mesoionic oxazolo-pyridazinones and methyl/ethyl propiolate. The mesoionic compounds were generated in situ by action of acetic anhydride on 3(2H)pyridazinone acids obtained from corresponding esters by alkaline hydrolysis followed by acidification. The structures of the compounds were confirmed by elemental analyses and IR, 1H-NMR, 13C-NMR, and X-ray diffraction data. The regioselectivity of cycloaddition was evidenced by NMR spectroscopy and confirmed by X-ray analysis. The compounds were evaluated for their cytotoxicity on plant cells (Triticum aestivum L.) and crustacean animal cells (Artemia franciscana Kellogg and Daphnia magna Straus). The results indicated that the tested compounds exhibited low toxicity on the plant cell (IC50 values higher than 200 µM), while on Artemia nauplii no lethality was observed. Daphnia magna assay showed that pyrrolo[1,2-b]pyridazines 5a and 5c could exhibit toxic effects, whereas, for the other compounds, toxicity was low to moderate. Also, the cytotoxic effects of the compounds were tested on three human adenocarcinoma-derived adherent cell lines (colon LoVo, ovary SK-OV-3, breast MCF-7). The in vitro compound-mediated cytotoxicity assays, performed by the MTS technique, demonstrated dose- and time-dependent cytotoxic activity for several compounds, the highest anti-tumor activity being observed for 5a, 2c, and 5f, especially against colon cancer cells. [ABSTRACT FROM AUTHOR]

HER2 gene, located in chromosome 17q12–q21, encodes a transmembrane protein of 185 kDa with tyrosine kinase activity and an important role in tumorigenesis. The presence of the Ile655Val polymorphism identified in transmembrane domain of the HER2 protein was observed in tumors with different locations. Previous studies have shown that changing the existing isoleucine (Ile) to valine (Val) at codon 655 of the HER2 gene causes an intensification of dimerization, autophosphorylation of the HER2 receptor, as well as tyrosine kinase activity, which influences cell growth and tumor proliferation. The aim of this study was to assess the HER2 codon 655 polymorphism related to tumor cells proliferation in urinary bladder cancer. Tumors obtained from patients with bladder cancer were analyzed by PCR-RFLP for the presence of polymorphism at codon 655 of the HER2 gene. The expression of HER2 and pHER2 proteins was determined immunohistochemically. The DNA content, from the tumor cells, was analyzed by flow cytometry for cell cycle phases and DNA ploidy. HER2 heterozygous genotype was dominant in carcinomas with co-expression of HER2 and pHER2. The tumors HER2 and pHER2 positive from Ile/Val genotype group had a higher level of S-phase fraction than those from Ile/Ile genotype group. The S-phase fraction was also elevated in heterozygous than homozygous tumor in patients with lymph node metastases. The HER2 heterozygous cases were aneuploid predominantly. The presence of Ile/Val genotype in bladder carcinoma seems to be associated with a high proliferation. These results may suggest that HER2 codon 655 polymorphism can be used in the evaluation of disease progression in bladder cancer. [ABSTRACT FROM AUTHOR]

Establishing the prognosis and the optimal treatment of cancer is of great importance in clinical practice. The choice of treatment should ideally be based on predictors of response to treatment, but many of the potential indicators still require clinical validation. Since cell proliferation plays an important role in determining tumor behavior, markers of cell proliferation may be useful for the prediction of prognosis and chemotherapy response. Therefore, the present study aims to determine and evaluate, in association with some clinical and histopathological prognostic factors, the nuclear protein Ki-67, widely used for the evaluation of tumor cell proliferation, and topoisomerase II alpha (Topo IIa), also a nuclear protein with similar expression pattern to Ki-67, considered more for its predictive potential value in response to chemotherapy, especially after association with Her2, in order to establish additional biomarkers that might be useful in order to improve treatment decisions and prognosis. However, there are currently no standardised staining techniques and clinically validated cutoff points for Ki-67, as well as for Topo IIa, so by using flow cytometry or immunohistochemistry analysis we determined higher levels of these parameters in poorly differentiated tumors; no association was observed with lymph node status and with patients’ age. Tumors with high Ki-67 index were also associated with an increased expression of Topo IIa. No obvious positive association was observed between Her2/neu protein expression and Topo IIa or Ki-67, but approximately half of Her2positive tumors overexpress Topo IIa and Ki-67. The determination of chemosensitivity in association with Topo IIα expression showed in tumor cell lines a correlation between the response to doxorubicin (IC50) and the level of protein expression of Topo IIα. Along with Ki-67, Topo IIα may be useful in assessing tumor aggressiveness, being important risk stratification tools, arguing the early application of a more aggressive treatment to the group of patients apparently at low risk. In addition, the increased expressions of these biological markers could be important predictive factors for chemosensitivity. [ABSTRACT FROM AUTHOR]

The aim of our study was to develop an in vitro, human experimental model, used in modulation experiments with treatments in which cytostatics and/or biologically active compounds were used, following the expression of soluble biomarkers associated with tumor proliferation processes and angiogenesis. As an experimental model, we used the MCF-7 breast cancer cell line, on which we applied treatments with cyclophosphamide, epirubicin, avastin and some flavonoids like resveratrol, quercetin and curcumin. We evaluated the expression of soluble biomarkers (VEGF, MMP-2, TIMP-1, cathepsin D, sialic acid) associated with the processes of cell proliferation and angiogenesis in breast cancer, which could be used in the cancer clinic to improve diagnostic procedures and prognosis, highlighting the risk of metastasis, and monitoring treatment. Cytotoxicity tests by the MTS or RTCA method indicated that epirubicin and cyclophosphamide treatments caused an increase in cell lysis percentages and thus a decrease in cell viability. The analysis of the data obtained in case of modulating the release of VEGF in the supernatants of the MCF-7 line treated with cytostatics and biocomposites showed that the combined treatment of cytostatics (epirubicin + cyclophosphamide), prolonged up to 48-72 hours, determined the decrease of VEGF levels. The present study showed that quercetin, administered before or after cytostatics, has the effect of decreasing the concentration of VEGF, with the prolongation of treatments at 72 hours, thus decreasing the angiogenesis. In case of resveratrol, the accentuated decrease in VEGF level occurs when the biocomposite is administered 24 hours before cytostatics. The analysis of the data obtained in case of modulating the release of MMP-2 in the supernatants of the MCF-7 line treated with cytostatics and biocomposites showed that the 24hour treatment with all three biocompounds studied decreased the levels of this proteinase, the same effect being noticed in cytostatics at 48 hours and 72 hours. The analysis of the data obtained in case of modulating the release of TIMP-1 in MCF-7 supernatants treated with cytostatics and biocomposites showed that the 48-72-hour treatments with resveratrol and curcumin caused the decreased levels of this molecule, the same effect being generally observed in combined treatments with cytostatics and biocomposites, especially for 72 hours. In case of quercetin, it seems that the maximum effect is obtained at the combined treatment in which the biocompound is added after cytostatics, except for the time of 72 hours. The combined treatments cause a decrease in both cathepsin D and sialic acid in the supernatants of the treated cells for a longer period, of 48-72 hours. As in case of cathepsin D, the sequential treatments with epirubicin + cyclophosphamide (24 hours) and avastin (24 or 48 hours) determine the decrease of sialic acid levels compared to avastin (48 hours) or epirubicin + cyclophosphamide (48 hours). The combinations of cytostatics and biocompounds are more effective with increasing treatment time, the best effect being obtained at 72 hours. There is a significant direct correlation between cathepsin D and sialic acid levels (p<0.001). The sequential treatment, with cytostatics and biocompounds, generally causes a decrease in the level of proliferation and angiogenesis biomarkers, especially for the 48- and 72-hour periods, compared to single treatments, but it depends on the biocompound and the time of administration (before or after cytostatics). [ABSTRACT FROM AUTHOR]