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

Comparison of the effects of three different Baccaurea angulata whole fruit juice doses on plasma, aorta and liver MDA levels, antioxidant enzymes and total antioxidant capacity

Purpose Baccaurea angulata (common names: belimbing dayak or belimbing hutan) is a Malaysian underutilized fruit. The preliminary work on B. angulata fruit juice showed that it possesses antioxidant properties. Therefore, further work is needed to confirm the efficacy and proper dosage of B. angulata as a potential natural antioxidant. The present study was thus carried out to compare the effects of three different B. angulata whole fruit (WF) juice doses administered at nutritional doses of 0.50, 1.00 and 1.50 ml/kg/day on plasma, aorta and liver malondialdehyde (MDA) levels, antioxidant enzymes (superoxide dismutase, glutathione peroxidase and catalase) as well as total antioxidant capacity in rabbits fed high-cholesterol diet. Methods Thirty-five male rabbits of New Zealand strain were randomly assigned to seven groups. For 12 weeks, group CH was fed 1% cholesterol diet only; group C1 was fed 1% cholesterol diet and 0.50 ml/kg/day B. angulata WF juice; group C2 was fed 1% cholesterol diet and 1.00 ml/ kg/day B. angulata WF juice; group C3 was fed 1

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

Anti-cancer activities of β-mangostin against oral squamous cell carcinoma

Oral squamous cell carcinoma (OSCC) is one of malignant tumors with poor prognosis resulting in major morbidity and mortality. The actual curative treatment is usually chemotherapy with concurrent radiation, sometimes combined with surgery. Unfortunately, the strength of the drugs used in chemotherapy causes side effects that can bring discomfort and inconvenience. Herbal remedies have been used for thousands of years with very minimal side effects and clearly merit extended research for their ability to selectively kill cancer cells. The genus of Garcinia is well known as a medicinal plant in Southeast Asia. β-mangostin, a xanthone from the pericarps of various species of Garcinia has been shown to exhibit anti-cancer activities in various human cancer cells. However, no attempt has been made to explore the potential benefits of this xantone for treatment and/or prevention of OSCC. Here, we report that β- mangostin exhibits anti-proliferative effect and induces apoptosis in human oral squamous cell carcinoma cell lines HSC-3 and Ca-922. MTT assay showed that β-mangostin markedly inhibited proliferation of HSC-3 and Ca-922 cells in a dose- and time-dependent manner. The apoptosis induced by β-mangostin was clearly detected by flow cytometry in both cell lines and confirmed by caspase activity assays. Moreover, quantitative RT-PCR revealed that apoptotic activity by β-mangostin in HSC-3 and Ca-922 cells is associated with an up-regulation of caspase - 8 and pro-apoptotic PUMA genes. These results identify β-mangostin as a potential therapeutic agent for human oral squamous cell carcinoma

Expression of collagenases matrix metalloproteinases and YB-1 oncogenic factor in malignant melanoma cancer cells and its regulation by stromal fibroblasts

Background and Objective: Fibroblast stromal cells actively participates in tumor invasion by secreting matrix metalloproteinases (MMPs) within the tumor microenvironment. Expression of these enzymes is primarily regulated at a transcriptional level via interaction with transcription factors. Among these factors, YB-1 oncogenic factor binds with different nucleic acids to exert its diverse influences. Also it represents an important prognostic indicator in many types of tumors. The aim of this study was to assess the expression of collagenases MMPs (MMP1, MMP8, MMP13) and the cellular proliferation in one of the most invasive types of cancer cell types which is the A375 malignant melanoma cancer cell line using co-culture settings. Also, the study attempted to assess the expression of YB-1 factor and its in vivo interaction with the AP-1 gene promoter sequence. Materials and Methods: The experiment involved growing A375 cells with CCD1079SK fibroblasts cells in co-culture environment. The proliferation of cells was determined using serial trypan blue assays, while the expression of YB-1, MMP1, MMP8 and MMP13 was determined by the use of real-time PCR and western blotting analysis. The potential interaction between YB-1 protein and AP-1 promoter sequence was assessed through chromatin immunoprecipitation (ChIP) assay. SPSS with independent t- test was used to compare cell proliferation and real-time PCR Ct mean values between samples. Results: In co-culture setting, the proliferation of A375 cancer cells was significantly faster than the cells in monoculture setting (5.1×105, 3×105 respectively in day 3) (p<0.05). Also, there was a significant increase in the expression of MMP1 enzyme. YB-1 and MMP8 were significantly expressed more in the A375 cancer cells in comparison with normal fibroblasts cells (p<0.05). Conclusion: The study confirms the role of stromal fibroblasts by enhancing the proliferation of melanoma cancer cells in vitro and increasing the expression of the MMP1 enzyme. In addition, YB-1 factor remains as an important prognostic indicator in cancer that might regulate expression of MMPs without binding to the AP-1 promoter sequence

YB-1 gene expression in A375 malignant melanoma cells

Background: YB-1 is a DNA and RNA binding protein that is involved in almost all DNA and mRNA related processes in cellular proliferation and differentiation. An increased expression of YB-1 is frequently being detected in different types of human cancers including breast, ovarian, thyroid and colorectal cancers. In addition, the elevated levels of YB-1 were found to be correlated with tumour progression and resistance to chemotherapy. Objectives/Research Problem: This study was conducted to measure YB-1 gene expression in A375 Malignant melanoma cell line by comparing its level with normal human fibroblasts cells. Materials and Method: These cell lines were derived from the American Type Cell Collection (ATCC), USA. The cells were then grown in Dulbecco’s Modified Eagle Medium supplied with 10% Foetal bovine serum, 1% of penicillin-streptomycin, 1% HEPES solution and 1% sodium pyruvate. After determination of the cell growth curve, the samples were collected for the exponential growth phase and the plateau growth phase. The samples RNA was then extracted and normalized for cDNA formation followed by reverse transcription PCR. The Cq values were determined for YB-1 protein, alpha tubulin and GAPDH. Results and Discussion: There was a 12.64-fold increase of YB-1 gene expression in the A375 cell line in comparison with normal fibroblasts cells; also, there was significant reduction in YB-1 gene expression in the plateau phase of growth in comparison with the exponential growth phase in the A375 cells (10.8-fold reduction in expression)

In vitro antibacterial effect of the leaf extract of Murraya koenigii on cell membrane destruction against pathogenic bacteria and phenolic compounds identification

Introduction: Different parts of Murraya koenigii are traditionally used for treating various infections. Multidrug-resistant bacteria is considered one of the most significant threats to public health and urgent attention is needed to address this pressing problem. The aim of this study was to examine the antibacterial effect of the leaves and stems of the plant to identify and screen for phenolic compounds and to explore whether the potent plant extract had a destructive effect on the cell membrane. Materials and methods: Leaf and stem extracts extracts (hexane, chloroform, ethyl acetate and methanol) of the plant were screened using micro broth dilution methods. Nine strains of bacteria were used in this study. The identification of phenolic compounds was performed using UPLC, and the membrane destruction was observed using SEM. Results: The ethyl acetate extract of the leaves gave the lowest MIC value (15.63 μg/mL) against S. aureus, E. coli 0157:H7. V. alginolyticus, V. parahaemolyticus and Y. enterocolitica. Among the tested compounds, the least MIC value of 31.25 μg/ml was observed from myricetin and quercetin against Y. enterocolitica (ATCC 23715). The two major peaks matched with quercetin and myricetin aglycons at the retention time of 3.853 and 4.160 min and two myricetin and quercetin derivatives at the retention time of 4.162 and 4.406. The alteration of the cellular morphology of the bacteria which comprises alteration in their arrangement, rigidity and destruction of the membrane was observed after treatment with plant extract. Conclusions: The findings of this research establish the efficacy of ethyl acetate leaf extract of this plant in treating diseases associated with pathogenic bacteri

Porcupine bezoar exhibit as anti-cancer activities on human breast adenocarcinoma cells (MCF-7)

Purpose: Currently there were many treatments available to treat cancer patients, for instance chemotherapy, radiation therapy, immune therapy, and many more. Unfortunately, advancement in cancer therapy has resulted in increasing numbers of survivors which left to deal with side effects of their treatments. Therefore, discovery of alternative treatment with minimal side effect is crucial. Thus, study on bezoar take place as there were no scientific studies about porcupine bezoar to this date in Malaysia. This lead to the premise that porcupine bezoar could have greater potential for the chemoprevention activities. Methods: MCF-7 cells were treated with bezoar for cell proliferation, cell cycle, apoptosis and qPCR analyses. Cell viability of treated and untreated MCF-7 was examined by Trypan Blue Exclusion method. The cellular morphology of MCF-7 cells was observed by phase contrast microscopy. Flow cytometry analyzed cell apoptosis with annexin V/ propidium iodide (PI) and cell cycle with PI. qPCR was run in detecting gene expression involved in the apoptosis and cell cycle arrest. Result(s): Based on the conducted study and experiments, it is evident that the bezoar treatment in human periodontal ligament cells didn’t show cytotoxicity effect. Upon treated with optimized concentration of bezoar for 24, 48 and 72 hours, it shows inhibition of proliferation by 45.5, 31.25 and 37.1% respectively, significantly apoptosis and cell cycle arrest occurs. Bezoar treatment also induced apoptotic-liked in morphological changes. Upon qPCR analysis, its indicated mcf7 undergo intrinsic pathway of apoptosis and arrest at G1/G0 as gene expression high in cdk2, cyclin D and p21. Conclusion(s): Porcupine bezoar can be alternative treatment for cancer with minimal side effects. Yet, furthermore insights need to be studied