Expression of Matrix Metalloproteinase-13 in Human Skin Melanoma Cancer Treated by Baccaurea angulata in vitro.

This study aims to explore the cytotoxicity effects of B. angulata whole fruits and berries as well as to identify the fruit’s probable role on the level of MMP-13 protein expressions in human cancer cells. Cytotoxicity effects of B. angulata were evaluated in vitro using skin melanoma (A375) through treatment with novel B. angulata fibers (whole fruit and berries) via direct contact method. The growth inhibitions of the samples were evaluated through Methylene Blue Assay (MBA) with incubation time of the fibers on cells at 24, 48 and 72 hours. The results showed significant inhibition of growth in all samples with B. angulata whole fruit exhibiting the highest level of inhibition at 72 hours (87.69 %) while its berries showed a reading of 86.16 %. Consequently, this study indicates that both whole fruit fibers and berries of B. angulata may have cytotoxic effects against human skin melanoma. The quantitative expressions of MMP-13 in A375 cell lines which were subjected to the specified amount of fibers were evaluated through ELISA analysis. The results showed no expression of MMP-13 proteins in A375 cells for both whole fruit fibers (-1217.9 pg/ml) and berries (-1222.9 pg/ml). Furthermore, the results of the ELISA analysis depict a probable regulative effect of the fibers toward MMP-13 protein expressions in cancer cells. Hence, it can be concluded that B. angulata fruit has the potential to be used as a new source of natural substitute for anticancer treatment. Moreover, further study is needed in order to find the specific bioactive compounds involved in the anticancer properties which may have been influential in the regulation of MMP-13 proteins that could be fundamental in future endeavors for prospective therapeutic applications

Effect of shRNA mediated silencing of YB-1 protein on the expression of matrix collagenases in malignant melanoma cell in vitro

Background and Objective: YB-1 is a transcription and oncogenic factor capable of binding to DNA and RNA performing versatile functions within normal and cancer cells. Some studies reported the binding of YB-1 with a collagenases gene promoter and influencing their expression. In addition, the role of YB-1 in malignant melanoma was not elucidated. Thus, in this study, the aim was to knock down the expression of YB-1 in A375 malignant melanoma cancer cell using the shRNA approach and study its effect on cancer cell proliferation, migration, and expression of collagenases. Methods: A375 malignant melanoma cell lines were grown in standard conditions and were transfected with three plasmids containing a retroviral pGFP-V-RS vector, two of them containing targeting sequences for YB-1 mRNA. The third plasmid contained a scrambled mRNA sequence as a negative control. Expression of YB-1 was validated using immune-fluorescence staining, RT-PCR and western blotting. The cancer cell proliferation was determined using MTT assay, serial trypan blue cell counting and cell cycle flow-cytometry analysis. Expression of collagenases (MMP1, MMP8, and MMP13) was evaluated using RT-PCR and western blotting analysis. In addition, a wound-healing assay was used to assess cell migration potential. Statistical analysis was performed using one-way ANOVA test with Bonferroni post hoc analysis to compare the quantitative results among samples. Results: The established silenced cell strains (P1 and P2) had nearly 70% knockdown in the expression of YB-1. These YB-1 silenced strains had a significant cell cycle-specific reduction in cell proliferation (p < 0.05 in serial cell counting and cell cycle flow cytometry analysis, p < 0.001 in MTT assay). In addition, YB-1 silenced strains had a remarkable reduction in cell migration potential. Expression of MMP13 was significantly reduced in YB-1 silenced strains. Conclusion: YB-1 oncoprotein is a promising target in the treatment of malignant melanoma. Silencing of this protein is associated with significant anti-proliferative, anti-invasive and MMP13 insulating properties in A375 malignant melanoma cancer cell lines

In vitro degradation study of novel HEC/PVA/collagen nanofibrous scaffold for skin tissue engineering applications

The aim of this study was focused on the degradation behavior of electrospun (hydroxyethyl cellulose/poly(vinyl) alcohol) HEC/PVA and HEC/PVA/collagen nanofibrous scaffolds, as a potential substrates for skin tissue engineering in two biologically related media: phosphate buffered solution (PBS) and Dulbecco's modified Eagle's medium (DMEM) for 12 weeks incubation period. The scaffolds were characterized at different degradation times by a series of analysis including pH changes of solutions, weight loss, swelling ratio, SEM, ATR-FTIR, DSC, TGA and mechanical properties. The results indicated that HEC/PVA/collagen scaffolds were exhibited slower degradation rate in both medium as compared to HEC/PVA blend nanofibers. All fibers displayed uneven and rough surfaces towards the final week of incubation in both PBS and DMEM solution. As degradation time increased, there were little changes in the chemical structure as determined by FTIR spectra while thermal studies revealed that the melting temperatures and crystallinity of scaffolds were slightly shifted to a lower value. Both HEC/PVA and HEC/PVA/collagen fibers showed significant decrease in Young's modulus and tensile stress over 12 weeks degradation. These results show that these nanofibrous scaffold demonstrate degradation behavior that meets the requirement as potential degradable biomaterials for dermal replacement

In-vitro cytotoxicity of Trigona itama honey against human lung adenocarcinoma epithelial cell line (A549)

Introduction: Many efforts have been made to identify natural alternatives to reduce the side effects of cytotoxic drugs in cancer treatment. With this in mind, the current study aimed to investigate the cytotoxicity effects of one of the multifloral Malaysian honey, Kelulut honey (Trigona itama), as a potential natural anticancer agent in stimulating apoptosis and cell cycle arrest to a human lung adenocarcinoma epithelial cell line (A549). Methods: The cells were treated with various concentrations of T. itama honey for 24, 48 and 72 hours. The cytotoxicity and cell viability were determined using trypan blue exclusion assay (TBEA) and flow cytometric analysis. Results: The moisture content in the analysed honey was 14.3 ± 0.8%, which was within the accepted international standard. The pH, electrical conductivity and proline content were 3.17 ± 0.02, 0.47 mS/cm - 0.55 mS/cm and 19.1 mg/kg - 20.2 mg/kg respectively. The findings demonstrated a significant dose and time-dependent inhibitory effect of T. itama honey with the maximum cytotoxic effects were observed at 72 hours with 20% concentration of T. itama honey, indicating 100% growth inhibition. Meanwhile, IC50 of T. itama honey treatment for A549 cells was determined as 0.62% v/v. Moreover, T. itama honey has a promising cytotoxic effect and proven capable of inducing cell cycle arrest at G2/M phase at 72 hours of exposure with IC50 concentration. Conclusion: This study provides a prefatory evidence on T. itama honey’s significant anticancer activity against human lung cancer cell lines

A facile synthesis method of hydroxyethyl cellulose-silver nanoparticle scaffolds for skin tissue engineering applications

Green porous and ecofriendly scaffolds have been considered as one of the potent candidates for tissue engineering substitutes. The objective of this study is to investigate the biocompatibility of hydroxyethyl cellulose (HEC)/silver nanoparticles (AgNPs), prepared by the green synthesis method as a potential host material for skin tissue applications. The substrates which contained varied concentrations of AgNO3 (0.4%–1.6%) were formed in the presence of HEC, were dissolved in a single step in water. The presence of AgNPs was confirmed visually by the change of color from colorless to dark brown, and was fabricated via freeze-drying technique. The outcomes exhibited significant porosity of > 80%, moderate degradation rate, and tremendous value of water absorption up to 1163% in all samples. These scaffolds of HEC/AgNPs were further characterized by SEM, UV–Vis, ATR-FTIR, TGA, and DSC. All scaffolds possessed open interconnected pore size in the range of 50–150 μm. The characteristic peaks of Ag in the UV–Vis spectra (417–421 nm) revealed the formation of AgNPs in the blend composite. ATR-FTIR curve showed new existing peak, which implies the oxidation of HEC in the cellulose derivatives. The DSC thermogram showed augmentation in Tg with increased AgNO3 concentration. Preliminary studies of cytotoxicity were carried out in vitro by implementation of the hFB cells on the scaffolds. The results substantiated low toxicity of HEC/AgNPs scaffolds, thus exhibiting an ideal characteristic in skin tissue engineering applications

Identification and quantification of quercetin, a major constituent of Artocarpus altilis by targeting related genes of apoptosis and cell cycle: in vitro cytotoxic activity against human lung carcinoma cell lines

Nine phenolic compounds were identified and quantified in Artocarpus altilia fruit. One of the main compounds was quercetin, which is the major class of flavonoids has been identified and quantified in pulp part of A. altilis fruit of methanol extract. The aim of this study was to evaluate in vitro cytotoxic assay. Inhibitory concentration 50% concentration was determined using trypan blue exclusion assay. Apoptosis induction and cell cycle regulation were studied by flow cytometric analysis. The expression of apoptosis and cell cycle-related regulatory genes were assessed by RT-qPCR study of the methanol extract of pulp part on human lung carcinoma (A549) cell line. A significant increase of cells at G2/M phases was detected (P<0.05). Furthermore, the pulp of the fruit downregulated the expression of antiapoptosis gene BCL-2 and upregulated the expression of pro-apoptosis gene BAX. CASPASE3 was also activated by the fruit, which started a CASPASE-3-depended mitochondrial pathway to induce apoptosis. As the results, the pulp was the most active in terms of all tests, due to high amount of quercetin in pulp part, 78% of total flavonoids. Taken together, these findings suggested that A. altilis induces apoptosis in a mitochondrial-dependent pathway by releasing and upregulating CYTOCHROME C expression and regulates the expression of downstream apoptotic components, including BCL-2 and BAX

Preliminary screening of cytotoxic properties of Baccaurea angulata in several human cancer cell lines in vitro

Fruits have been acknowledged as a reservoir for numerous bioactive compounds. Baccaurea angulata, belonging to the Euphorbiaceae family, has been reported to contain several nutritional properties such as proteins and carbohydrates. Presently, B. angulata has not been fully exploited as little is known regarding its scientific properties. This preliminary study aims to explore the cytotoxicity effects of various extracts and fibers of B. angulata on human cancer cells. Cytotoxicity effects of B. angulata were evaluated on cervix cancer (HeLa) and skin melanoma (A375) cells through treatment with hexane, methanol and dichloromethane extracts of B. angulata as well as with novel B. angulata fibers (whole fruit and berries) via direct contact method. The growth inhibitions of the samples were evaluated through Methylene Blue Assay (MBA), with incubation time of 24, 48 and 72 hours. From the results, it was observed that for HeLa cells, berries showed the highest growth inhibition (90.69 μg/mL, 72 hours). The solvent-based extracts were found to be cytotoxic towards HeLa cells with methanol extracts exhibiting the highest cytotoxicity with EC50 of 17.21 μg/mL. For A375 cells, all extracts also exhibited cytotoxic effects, where the highest inhibition activity was induced by hexane extracts (89.40 μg/mL, 72 hours). Consequently, the study indicates that B. angulata fruit may have significant growth inhibitory and cytotoxicity effects toward HeLa and A375 cells. It can be presumed that B. angulata fruit has the potential to be used as a new alternative resource that could be fundamental in future endeavors for prospective therapeutic treatments for cancer

Expression of matrix metalloproteinase-13 in human skin melanoma cancer treated by baccaurea angulata in vitro

Currently, B. angulata has not been fully exploited as little is known regarding the scientific and anti-cancer properties of the fruit. Also, the degree of Matrix Metalloproteinase-13(MMP-13) association in the development and metastasis of cancer remains to be scantily know

Expression of matrix metalloproteinase-13 in human skin melanoma cancer treated by Baccaurea angulata in vitro

This study aims to explore the cytotoxicity effects of B. angulata whole fruits and berries as well as to identify the fruit’s probable role on the level of MMP-13 protein expressions in human cancer cells. Cytotoxicity effects of B. angulata were evaluated in vitro using skin melanoma (A375) through treatment with novel B. angulata fibers (whole fruit and berries) via direct contact method. The growth inhibitions of the samples were evaluated through Methylene Blue Assay (MBA) with incubation time of the fibers on cells at 24, 48 and 72 hours. The results showed significant inhibition of growth in all samples with B. angulata whole fruit exhibiting the highest level of inhibition at 72 hours (87.69 %) while its berries showed a reading of 86.16 %. Consequently, this study indicates that both whole fruit fibers and berries of B. angulata may have cytotoxic effects against human skin melanoma. The quantitative expressions of MMP-13 in A375 cell lines which were subjected to the specified amount of fibers were evaluated through ELISA analysis. The results showed no expression of MMP-13 proteins in A375 cells for both whole fruit fibers (-1217.9 pg/ml) and berries (-1222.9 pg/ml). Furthermore, the results of the ELISA analysis depict a probable regulative effect of the fibers toward MMP-13 protein expressions in cancer cells. Hence, it can be concluded that B. angulata fruit has the potential to be used as a new source of natural substitute for anticancer treatment. Moreover, further study is needed in order to find the specific bioactive compounds involved in the anticancer properties which may have been influential in the regulation of MMP-13 proteins that could be fundamental in future endeavors for prospective therapeutic applications