scholarly journals Evaluation the Effect of Natural Compounds: Vitamin C, Green Tea, and their Combination on Progression of Mg-63 Osteosarcoma Cell Line Cells. (An In Vitro Study)

2021 ◽  
Vol 9 (A) ◽  
pp. 1277-1284
Author(s):  
Hiam Rifaat Hussien Mohammed ◽  
Amr Helmy Moustafa El Bolok ◽  
Sherif Farouq Elgayar ◽  
Maii Ibrahim Ali Sholqamy

BACKGROUND: Osteosarcoma (OS) is considered extremely rare type of bone tumor although it is the most common type of malignant bone tumor in children with less common occurrence in elderly patients. Herbal plants and phytoconstituents are recently used in the treatment of OS to avoid the side effects of chemotherapeutic drugs. AIM: The aims of the present study are to investigate the effect of natural compound Vitamin C, green tea, and their combination on OS cell line (Mg-63 cells) after 72 h. MATERIAL AND METHODS: Mg-63 cells were obtained from Nawah scientific and divided to four groups: Control untreated cells, Vitamin C treated group, green tea treated group, and Vitamin C and green tea treated group (compounds combination treated group). The viability of treated cells was examined by sulforhodamine B (SRB) assay. Antioxidant 2,2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH) assay was performed to investigate the antioxidant property of Vitamin C, green tea, and their combination. Flow cytometer analysis was applied to demonstrate cell cycle analysis and apoptosis. Wound width and cell migration were calculated by wound healing assay. RESULTS: SRB cytotoxic assay revealed that the Vitamin C, green tea, and their combination have a cytotoxic effect on MG-63 cells and Vitamin C has more cytotoxic effect than other two groups. Antioxidant DPPH assay showed that Vitamin C is more antioxidant agent than green tea and their combination on MG-63 cells. Flow cytometry assay revealed that the all-treated cells in different groups are arrested in cell cycle. Vitamin C, green tea, and their combination induced apoptosis and necrosis. Migration of MG-63 cells is inhibited after treated by Vitamin C, green tea, and their combination. CONCLUSION: Vitamin C, green tea, and their combination have cytotoxic effect on Mg-63 cells, also induced their effects on the cell cycle distribution and apoptosis. Anti-oxidant test was applied on three drugs revealed the powerful anti-oxidant capacity of Vitamin C than green tea and their combination. At least wound healing test was applied on malignant Mg-63 cells treated with our drugs that revealed Vitamin C was more effective.

2020 ◽  
Author(s):  
Kim Fey Leu ◽  
Menaga Subramaniam ◽  
Xinghua Wang ◽  
Zao Yang ◽  
Lee Fah Yap ◽  
...  

Abstract Background Nasopharyngeal carcinoma (NPC) is among the most common head and neck malignancies seen among adults in Malaysia. Therefore, discovery of novel anti-cancer herbal drugs is of importance. In this study, the cytotoxic effect was conducted on a traditional Chinese herbal prescription (Xiao Xian Xiong Decoction (XXXD) that is made up of 3 Chinese herbal medicines, namely Huanglian (Coptidis Rhizome), Banxia (Pinellia Rhizome), Gualuo (Fructus Trichosanthis).Methods The cytotoxic effect of the individual herb and in combination of two and three herbs was studied on 8 nasopharyngeal cancer cell lines. Global gene expression analysis was carried on extracted RNA using nCounter XT Gene Expression Assay.Results TWO-1, TWO-4, HONE-1, SUNE-1, CNE-2, HK-1, CNE-1 and C666-1 treated with Huanglian, the IC50 values obtained were 24.48, 11.77, 4.48, 10.72 6.32, 11.10, 6.77 and 27.30 µg/ml, respectively. For combination of Huanglian and Banxia, the IC50 values obtained were 74.09 µg/ml (TWO-1), 25.80 µg/ml (TWO-4), 38.10 µg/ml (HONE-1), 29.46 µg/ml (SUNE-1), 19.0 µg/ml (CNE-2) and 20.12 µg/ml (HK-1) but did not exert 50% cell killing in CNE-1 and C666-1 cell lines. The IC50 value attained for the combination of Huanglian and Gualuo was 40.70 µg/ml in HONE-1 cell line. The IC50 values obtained for XXXD (triple combination of Huanglian, Banxia and Gualuo)-treated in HONE-1 and CNE-2 cell lines were 88.55 and 92.42 µg/ml, respectively. Out of all these 7 groups of herbal samples, Huanglian showed the highest cytotoxicity against 8 NPC cell lines with the lowest IC50 value of 4.48 µg/ml recorded in HONE-1. Global gene expression showed Huanglian significantly downregulated genes associated with cell cycle arrest and apoptosis, and thus inhibit HONE-1 cell growth.Conclusions This study suggest that Huanglian could be a potent anticancer herb targeting HONE-1 cancer cell line.


QJM ◽  
2021 ◽  
Vol 114 (Supplement_1) ◽  
Author(s):  
Walaa Adel Abd El Moez Ahmed ◽  
Seham Hassan Refaat ◽  
Hany Waheeb Abd El-Malak ◽  
Asmaa Ibrahim Ahmed ◽  
Ashraf Mohammed Mostafa Sadek

Abstract Background Cyclophosphamide (CYP) is considered one of the most successful chemotherapeutic drugs involved in anticancer regimens. However, it has multiple side effects. Mesna has an antiinflammatory effect and usually used in the treatment of cystitis. Vitamin C is a water-soluble vitamin which has a potent anti –oxidant effect that might protect cells against the oxidative damage caused by cyclophosphamide. Aim of the study The aim of the present study was comparing between the possible protective effect of vitamin C versus mesna and their combined therapy against the histological and immunohistochemical changes induced by cyclophosphamide on the urinary bladder of adult male albino rats. Material and methods Thirty adult male albino rats were divided into 5 groups, 6 rats each; (control(?), CYP-treated group (Пa), recovery group(Пb), mesna-treated group(???), vitamin C- treated group(?V) and the combined group (V). Histological examination of the H&Eand toluidine blue stained sections was done by light microscopy to assess the changes in the architecture of the urinary bladder. Avidin Biotin staining was performed for demonstration of iNOS immunoreactivity and histomorphometric analysis was done. Results Examination of H&E stained sections of cyclophosphamide- treated group (Пa) showed variable degrees of urothelial affection. Wide areas of urothelial cell degeneration with evident basal cytoplasmic vacoulatins, surface erosions and sloughed urothelial debris. Other Areas showed surface ulceration, completely denuded urothelium or the presence of multiple cysts replacing the urothelium and resting on the basement membrane. Semithin sections showed that the cytoplasmic microvesicles of umbrella cells were hardly detected. The Avidin Bioton stained sections showed intense positive immune reaction to iNOS in all layers of the urothelium. Scanning electron microscopy showed loss of the normal polygonal shape of the superficial epithelial cells, erosions, or deep ulcerations. Moreover, examination of the lamina propria by light microscopy showed multiple mononuclear inflammatory cells were detected, mast cells were seen in the lamina propria and some of them were invading the basement membrane of the urothelium. Dilated blood vessels and wide areas of extravasted blood (hemorrhage) were also observed. In addition, multiple epithelial cell nests of irregular shapes and sizes were deeply located in the lamina propria and exhibited pale esinophilic colloid discharge in their lumen. Scanning electron microscopy showed dense deposition of collagen fibers in both superficial and deep fibers of the lamina propria. Minimal improvement was observed in the recovery group (subgroup Пb). Mild to moderate improvement of the previous findings of CYP treated group was observed with each of mesna and vitamin C. Combined treatment of CYP with both of mesna and vitamin C induced apparent restoration of almost of the normal architecture of the urinary bladder. Conclusion CYP consumption developed morphologic and morphometric changes in the urinary bladder. The recovery group showed minimal improvement of the bladder architecture and increasing the period of recovery might produce better results. Each of vitamin C and mesna- treated groups induced mild to moderate improvement on the bladder architecture but treatment with combination of both of them offered remarkable improvement. Combined mesna and vitamin C induced significant protection via their combined anti-inflammatory and anti-oxidant proprieties.


Blood ◽  
2016 ◽  
Vol 128 (22) ◽  
pp. 1624-1624 ◽  
Author(s):  
Chandrima Sinha ◽  
John Bowling ◽  
Aman Seth ◽  
Bensheng Ju ◽  
Bhaskar Kahali ◽  
...  

Abstract The marine environment has been shown to be a rich source of pharmacologically-active secondary metabolites. Three marine- compounds have FDA approval for cancer indications. Aaptamine is a sponge-derived alkaloid that exhibits multiple pharmacological activities including proapoptotic/antiproliferative effects on leukemia cell lines. The effect of the aaptamine class has not been previously studied for high risk leukemias with mixed-lineage leukemia (MLL) gene rearrangements. Using the CellTiter-Glo cell viability assay we evaluated the cytotoxic effect of aaptamine against a panel of leukemia cell lines. We observed that cell lines containing t(4;11) are the most sensitive to aaptamine. Translocation (4;11) is associated with mixed-lineage leukemia and responsible for a very aggressive and refractory pediatric leukemia. Specifically, infants less than one year with t(4;11) have poor survival rates (≈ 19%) and new therapies are urgently needed. Interestingly other MLL cell lines that contain t(9;11) are comparatively less susceptible to aaptamine-mediated cytotoxicity. Jurkat cells overexpressing MLL-AF4 fusion protein are also more sensitive to aaptamine-induced cytotoxicity than wild type or MLL-AF9 overexpressing Jurkat cells indicating the specificity of aaptamine for t(4;11). To further confirm the specificity we conducted a flow based apoptosis assay and observed that aaptamine induces significant apoptosis and necrosis in RS4;11 and MV4;11 cell lines starting at 10µM but not in the t(9;11) containing THP1 cell line. We also found that aaptamine treatment induced G0/G1 arrest specifically in t(4;11) containing cell lines but not in THP1. Additionally we observed that aaptamine did not induce any resistance to the sensitive cell lines after 27 days of chronic exposure. Importantly the compound was well tolerated by healthy activated PBMCs and mice at high concentrations. In order to decipher the mechanism of specificity, we conducted a global proteomic study with treated and untreated RS4;11 and THP1 cell lines. Our proteomic data revealed a significant upregulation of p21 and p27 in aaptamine treated RS4;11 cells but not in THP1. In agreement with the proteomic data, we observed a dose-dependent upregulation of p21 and p27 in both protein and mRNA levels in RS4;11 and MV4;11 cells but not in resistant THP1 cells. Using p21 and p27 promoter-driven luciferase reporter constructs, we observed a significant upregulation of luminescence signal in the RS4;11 cell line at much lower concentration of aaptamine (1µM) whereas the THP1 cell line required 50µM of aaptamine for significant increase in luminescence signal. Cyclin-dependent kinase regulates the G1/S cell cycle transition by phosphorylating retinoblastoma protein (RB). Upregulation of cyclin-dependent kinase inhibitors, such as p27 and p21, promote RB hypophosphorylation and induce G0/G1 arrest. To confirm that this molecular mechanism is responsible for aaptamine induced G0/G1 arrest, we investigated the effect of aaptamine on Rb phosphorylation. We observed a dose dependent downregulation of Rb phosphorylation by aaptamine in sensitive cell lines and predicted it as a major cause of cell cycle arrest. Previous studies have shown that translocation (4;11) is associated with p27 upregulation; thus we hypothesize by further upregulating p27, aaptamine may induce G0/G1 arrest specifically in t(4;11) containing cell lines. To validate the efficacy of aaptamine in vivo, we xenografted 10 NSG mice with 1 million luciferase expressing RS4;11 cells. Four days after leukemia induction we treated half of the mice with subcutaneous injection of aaptamine (100mg/kg, daily) and the other half received vehicle treatment. Bioluminescence imaging (BLI) data revealed a significantly lower disease (p< 0.03) burden in the aaptamine treated group compared to vehicle treated group after 2 weeks. These findings are being confirmed in patient samples. Additional aaptamine analogs are being designed and will be evaluated for improved therapeutic efficacy. Together our in vitro and in vivo findings suggest that by inducing p21 and p27 aaptamine can induce cell cycle arrest and eventually apoptosis specifically in leukemia cells that contain t(4;11) with relatively low toxicity . Therefore the aaptamine class of drug may provide additional therapeutic options for t(4;11) containing high-risk MLL leukemia patients. Disclosures No relevant conflicts of interest to declare.


2019 ◽  
Vol 5 (4) ◽  
pp. 145-150
Author(s):  
Uyovwiesevwa Ataihire Johnson ◽  
◽  
Eze Kingsley Nwangwa ◽  
John Chukwuka Igweh ◽  
◽  
...  

Antioxidants are specialized macro-molecules that neutralize harmful substances; free radicals. These radicals supposedly harm tissues, destroy food items, and damage materials. In living organisms, antioxidants can take the form of enzymes, and may be regularly added to oils, metals, foodstuffs, as well as numerous other materials to mitigate the damaging effect of free radical. Current study was designed to investigate the biochemical changes in antioxidant enzyme activities, following administration of Silybum marianum (an ancient medicinal plant of the Carduus marianum family) on Alloxan-Induced, diabetic rats. One hundred and twenty-five (125) rats were procured, made to acclimatize for two weeks, and then randomly grouped into five (5) groups of (n=25). Group 1: Non-Diabetic (Control) rats, Group 2 diabetic untreated rats, while groups 3, 4 and 5 comprised of vitamin-C treated rats (diabetic), Silymarin (extract), and Vitamin C + Silymarin (extract) combined treatment respectively. After four weeks of treatment with test extract, animals were then sacrificed, and blood samples collected and assayed for biochemical [anti-oxidant] enzyme activity. Upon statistical analysis, one way Analysis of variance (ANOVA) showed Catalase (CAT), superoxide dismutase (SOD) and malonaldehyde (MDA) activities to have significantly decreased for extract + vitamin C treated group (Group V) when compared with control (Group I). It was also noted that the use of the combined antioxidants vitamin C and silymarin resulted in a significant reduction in ROS production with decreased SOD and CAT enzyme activities. It is therefore likely that, improvements in antioxidant enzyme activities are a function of extract and/or Vitamin C administration to animals. Thus, Silymarin has antioxidant and regenerative potentials to damaged tissues.


2001 ◽  
Vol 20 (8) ◽  
pp. 418-425 ◽  
Author(s):  
C N Smith ◽  
C D Lindsay

Epidermal melanocytes have a higher sensitivity to sulphur mustard (HD) compared with other skin cell types.1 This may be due to the enzymatic production of melanin precursors exerting an additional cytotoxic effect following HD depletion of the cellular protectant, GSH.2,3 Stimulation of the protein kinase C pathway in melanocytes is known to increase melanin production in melanocytes and melanoma cell lines.4,5 In order to investigate the role of pigment synthesis in HD toxicology, cultures of an unpigmented melanoma cell line (C32) and of a pigmented melanoma line (G361) were treated with the potent diacyl glycerol analogue, oleoyl acetyl glycerol (OAG), in order to determine if protein kinase C-mediated increases in pigment production could increase sensitivity to subsequent HD exposure. Stimulation of C32 cells with OAG exerted a significant protective effect against the cytotoxic effects of HD. However, this was not due to increased melanin synthesis because this cell line cannot synthesize melanin pigments. The protective action observed is postulated to be due to modulation of protein kinase C activity. In contrast, stimulation of G361 melanoma cells with OAG resulted in an increased level of cytotoxicity upon subsequent exposure to HD. Protein kinase C controls several cellular pathways including checkpoints in the cell cycle, stalling the cell in G1 and promoting transition through the G2 /M boundary. Given the genotoxic properties of HD, these two points in the cell cycle are important in determining the overall cytotoxic effect of HD. Control of the cell cycle by protein kinase C modulation and manipulation of melanin synthetic pathways may have therapeutic benefits.


Proceedings ◽  
2019 ◽  
Vol 40 (1) ◽  
pp. 18
Author(s):  
Kubra Erdogan ◽  
Onur Eroglu

Glioblastoma multiforme (GBM) is a type of cancer which has the highest mortality rate among brain cancers (1–2). Momordica charantia, known as bitter melon, is a plant its pharmacological activities and nutritional properties. Due to contains bioactive compounds, M. charantia is used for cancer treatments, inflammation-related diseases and diabetes (3–4). In this study, it was aimed to investigate the effects of M. charantia extract on cell viability, cytotoxicity and migration capacity in U87G cell line. U87G was cultured in DMEM-high glucose containing FBS 10% (v/v) and penisillin-streptomicin 1% (v/v). Cells were incubated at 37 °C in a humidified 5% CO2 incubator. The cytotoxic effect of M. charantia extract was determined by MTT analysis, cell viability by survival analysis and migration by wound-healing analysis. The results were evaluated by using ANOVA and GraphPad Prism7.0 program (GraphPad Software, La Jolla, CA, USA) in three replicates. IC50 value of M. charantia extract was found 750 μg/mL which is statistically significant (* p < 0.05). The extract had an increasing lethal effect at the 16.6% (24 h), 42.6% (48 h), 79.3% (72 h) and 91.6% (96 h). According to the wound-healing analysis, the wound closed at 24 h in the control group and the wound gradually increased depending on time in the extract treated group. According to the results, M. charantia extract has a cytotoxic and a significant anti-proliferative effect on U87G. It might be used as therapeutic agent against to GBM. However, in order to understand the effect of M. charantia in living organisms, in vivo experiments must be determined.


Molecules ◽  
2021 ◽  
Vol 26 (18) ◽  
pp. 5653
Author(s):  
Grzegorz Adamczuk ◽  
Ewelina Humeniuk ◽  
Magdalena Iwan ◽  
Dorota Natorska-Chomicka ◽  
Kamila Adamczuk ◽  
...  

Leflunomide, an anti-inflammatory agent, has been shown to be effective in multiple myeloma (MM) treatment; however, the mechanism of this phenomenon has not been fully elucidated. The aim of the study was to assess the role of mitochondria and dihydroorotate dehydrogenase (DHODH) inhibition in the cytotoxicity of leflunomide in relation to the MM cell line RPMI 8226. The cytotoxic effect of teriflunomide—an active metabolite of leflunomide—was determined using MTT assay, apoptosis detection, and cell cycle analysis. To evaluate DHODH-dependent toxicity, the cultures treated with teriflunomide were supplemented with uridine. Additionally, the level of cellular thiols as oxidative stress symptom was measured as well as mitochondrial membrane potential and protein tyrosine kinases (PTK) activity. The localization of the compound in cell compartments was examined using HPLC method. Teriflunomide cytotoxicity was not abolished in uridine presence. Observed apoptosis occurred in a mitochondria-independent manner, there was also no decrease in cellular thiols level. Teriflunomide arrested cell cycle in the G2/M phase which is not typical for DHODH deficiency. PTK activity was decreased only at the highest drug concentration. Interestingly, teriflunomide was not detected in the mitochondria. The aforementioned results indicate DHODH- and mitochondria-independent mechanism of leflunomide toxicity against RPMI 8226 cell line.


2004 ◽  
Vol 67 (2) ◽  
pp. 337-351 ◽  
Author(s):  
James M. Jamison ◽  
Jacques Gilloteaux ◽  
M.Reza Nassiri ◽  
Meenakshi Venugopal ◽  
Deborah R. Neal ◽  
...  

2019 ◽  
Author(s):  
Siti Munirah Md Noh ◽  
Siti Hamimah Sheikh Abdul Kadir ◽  
Sushil Kumar Vasudevan

Anti-fibrotic properties of ranibizumab have been well documented. As an antagonist to Vascular Endothelial Growth Factor (VEGF), ranibizumab works by binding and neutralizes all active VEGF, thus limits the progressive cell growth and proliferation. Its application in ocular diseases has shown remarkable desired effects, however to date its anti-fibrotic mechanism is not well understood. In this study, we identified metabolic changes in ranibizumab-treated Human Tenos’s fibroblast (HTF). Cultured HTFs were treated for48 hours with 0.5 mg/ml of ranibizumab and 0.5 mg/ml control IgG antibody which serves as a negative control. Samples from each group were injected into Agilent 6520 Q-TOF Liquid Chromatography/ Mass Spectrometer (LC/MS) System to establish the metabolite expression in both ranibizumab treated cells and control group. Data obtained was analysed using Agilent Mass Hunter Qualitative Analysis software to identify the most regulated metabolite following ranibizumab treatment. At statistical analysis of p-value < 0.01 with the cut off value of two-fold change, 31 identified metabolites were found to be significantly up-regulated in ranibizumab-treated group, with 6 of the mostly up-regulated have insignificant role in fibroblast’s cell cycle and wound healing regulations Meanwhile, 121 identified metabolites were down-regulated with seven of the mostly down-regulated are significantly involved in cell cycle and proliferation. Our findings demonstrated that ranibizumab abrogates the tissue scarring process and wound healing formation by regulating the expression of metabolites associated with fibrotic activity. In particular, we found that vitamin Bs are important in maintaining normal folic acid cycle, nucleotide synthesis, homocysteine and spermidine metabolism. This study provides an insight of ranibizumab mechanism of actions on HTFs from the perspective of metabolomics.


2019 ◽  
Author(s):  
Siti Munirah Md Noh ◽  
Siti Hamimah Sheikh Abdul Kadir ◽  
Sushil Kumar Vasudevan

Anti-fibrotic properties of ranibizumab have been well documented. As an antagonist to Vascular Endothelial Growth Factor (VEGF), ranibizumab works by binding and neutralizes all active VEGF, thus limits the progressive cell growth and proliferation. Its application in ocular diseases has shown remarkable desired effects, however to date its anti-fibrotic mechanism is not well understood. In this study, we identified metabolic changes in ranibizumab-treated Human Tenos’s fibroblast (HTF). Cultured HTFs were treated for48 hours with 0.5 mg/ml of ranibizumab and 0.5 mg/ml control IgG antibody which serves as a negative control. Samples from each group were injected into Agilent 6520 Q-TOF Liquid Chromatography/ Mass Spectrometer (LC/MS) System to establish the metabolite expression in both ranibizumab treated cells and control group. Data obtained was analysed using Agilent Mass Hunter Qualitative Analysis software to identify the most regulated metabolite following ranibizumab treatment. At statistical analysis of p-value < 0.01 with the cut off value of two-fold change, 31 identified metabolites were found to be significantly up-regulated in ranibizumab-treated group, with 6 of the mostly up-regulated have insignificant role in fibroblast’s cell cycle and wound healing regulations Meanwhile, 121 identified metabolites were down-regulated with seven of the mostly down-regulated are significantly involved in cell cycle and proliferation. Our findings demonstrated that ranibizumab abrogates the tissue scarring process and wound healing formation by regulating the expression of metabolites associated with fibrotic activity. In particular, we found that vitamin Bs are important in maintaining normal folic acid cycle, nucleotide synthesis, homocysteine and spermidine metabolism. This study provides an insight of ranibizumab mechanism of actions on HTFs from the perspective of metabolomics.


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