The role of polyamines in beta vulguris (beetroot), zizhipus jujube (jujube) and vitis venefera (raisin) on human lung adenocarcinoma cells (A549) proliferation and apoptosis

Introduction: Polyamines are vital in maintaining human health because they are crucial for cell growth. Excessive intracellular polyamines in diet may promote unwarranted cell proliferation. Objective: This study aimed to investigate polyamines as a chemopreventive agent in selected fruits against human lung adenocarcinoma cells, A549. Materials and Methods: Beta vulgaris (beetroot), Ziziphus jujube (jujube) and Vitis vinifera (raisin) were freeze dried and kept at -80 °C until analysis. The anti-proliferative activity of selected fruits was assessed on A549 cells using MTT assay and the effect on A549 growth was completed via Trypan bluedye exclusion assay. Cell cycle profile and apoptosis were conducted with flow cytometer, while caspase assay was done using microtiter plate-reader. The quantification of intracellular polyamines was done using HPLC and the effect of these medicinal plants on the gene expression of selected polyamine metabolic enzymes was determined using qPCR. Results: The IC50 ranged between 15 to 30 mg/ml for these three fruits. A549 growth indicated significant decreased in cell viability after 48 h. Annexin-V/PI demonstrated significant increase in early apoptosis in beetroot treated cells while other fruits caused significant increase in late apoptosis at 48 h of treatments. There was a significant downregulation of ODC gene in cells treated with jujube and raisins while upregulation of SSAT gene in beetroot treated cells. Conclusion: These medicinal fruits are the promising candidates for food-based chemoprevention strategy for their low polyamine content, vital in modulating polyamine metabolism and promoting cell death

The role of polyamines as a chemopreventive agent in selected prophetic fruits against human lung adenocarcinoma cell line, A549

Polyamines are vital in maintaining human health because they perform certain functions that are necessary for cell development. These biogenic amines are naturally produced by the body but may also acquire through diet. Increased in intracellular polyamine promote unwarranted cell proliferation and to a certain extent, stimulates cancer initiation. Thus, it is crucial to evaluate polyamines content in medicinal fruits that claimed to cure cancer in order to avoid these unnecessary cell growth. This study aimed to investigate polyamines as a chemopreventive agent using selected prophetic fruits on human lung adenocarcinoma cells, A549. Five prophetic fruits were selected, including Phoenix dactylifera (ajwa dates), Beta vulgaris (beetroot), Ficus auriculata (fig), Ziziphus jujube (jujube) and Vitis vinifera (raisin). Polyamines concentration in selected prophetic fruits showed significant highest in jujube while lowest in fig.MTT assay suggested IC50 was ranged from 15 mg/ml to 30 mg/ml. Growth effect analysis indicated significant reduction in cell number after 48 h of treatments while cell viability decreased following 72 post-treatment. Total elimination of intracellular spermidine and spermine were observed in treated A549 cells as well. Expression of ODC and SSAT genes indicated significant downregulation of ODC gene in cells treated with ajwa, jujube and raisins while upregulation of SSAT gene in beetroot and fig treated cells. Cell cycle profile displayed cell cycle arrest at G2/M after 48 h of treatments in all treated cells. Annexin-V results demonstrated significant increase of early apoptosis in beetroot treated cells while other fruits caused significant increase in late apoptosis at 48 h of treatments. Caspase assay revealed significant activation of caspase 3, 8 and 9 in beetroot treated cells while no caspase activation was identified in other prophetic fruits treated cells. It was concluded that nutritional cancer therapy and preventive approaches for cancer using selected prophetic medicinal fruits are promising

The effects of Ficus auriculata (fig) on cell cycle profile and apoptosis activity of Human lung adenocarcinoma cells, A549

Ficus auriculata (fig) has an immense value of benefits with regards to their medicinal and therapeutic properties. (Fig) has been long used in traditional folk medicine, Traditional Chinese Medicine (TCM) and Ayurveda. Many scientific researches have proven the usage of this natural medicine in in vitro and in vivo studies, where anti-cancer is among of its recognized properties. To acquire further details on how F.auriculata (fig) inhibited cancer growth, we investigated the effects of its exposure on cell cycle profile. Subsequently we determined the type of cell death and the mechanism it induced using flow cytometer. Human lung adenocarcinoma cell line A549 was selected as the model of this study. Results indicated significant decreased of A549 cells in G0/G1 and concurrent accumulation of cells in G2/M phase. Cell death analysis revealed significant late apoptosis induced by F.auriculata (fig) via caspase independent pathway. Therefore, we concluded that F.auriculata (fig) is one of the promising natural medicines that can be used as cancer preventive strategy, as a result from its effects against cell cycle profile and apoptosis via caspase independent pathway

Ficus auriculata (fig) extracts induced cell cycle profile changes and apoptosis through caspase-independent pathway in human LungAdenocarcinoma cell line, A549

Background: Ficus auriculata (fig) has immense value of benefits with regards to their medicinal and therapeutic properties. It has been long used in traditional folk medicine, and one of the fruits mentioned in Al-Quran. Many scientific researches have proven the usage of this natural medicine in in vitro and in vivo studies, where anti-cancer is among of its recognized properties. Objective: To acquire further details on how F. auriculata (fig) inhibited cancer growth, we investigated the effects of its exposure on cell cycle profile. Methods: Subsequently we determined the type of cell death and the mechanism it induced using flow cytometer. Human lung adenocarcinoma cell line A549 was selected as the model of this study. Results: It is indicated significant decreased of A549 cells in G0/G1and concurrent accumulation of cells in G2/M phase. Cell death analysis revealed significant late apoptosis induced by F. auriculata (fig) via caspase independent pathway. Conclusion: Therefore, we concluded that F. auriculata (fig) is one of the promising natural medicines that can be used as cancer preventive strategy, as a result from its effects against cell cycle profile and apoptosis via caspase independent pathway

Polyamines deficient diet is a complementary strategy in fighting cancer: analysis on selected medicinal fruits

Polyamines are one of the oldest substances known in biochemistry consist of putrescine, spermidine and spermine. They are essential components of all living cells primarily involve in cell growth and proliferation. Upregulation of polyamines in human body is highly associated with various diseases, including cancer. Therefore, polyamines deficient diet (PDD) has become one of the strategies to inhibit carcinogenesis. This study was aimed to determine the polyamines content in selected anticancer fruits and their effect on polyamine polyamine metabolic enzymes was explored in human lung adenocarcinoma cells (A549). In this study, the selected medicinal fruits are Phoenix dactylifera (ajwa dates),Ficus auriculata (fig),Beta vulgaris (beetroot), Ziziphus jujube (jujube) and Vitis vinifera (raisin). The selected medicinal fruits were initially grouped into classes based on total polyamines content using High performance Liquid Chromatography (HPLC). The ability of selected fruits to reduce A549 intracellular polyamines was determined using the same technique. Accordingly, changes in polyamine metabolic enzymes activities; ornithine decarboxylase (ODC) and spermidine/spermine N-acetyltransferase (SSAT) were measured correspondingly using quantitative polymerase chain reactions (qPCR). Results suggested Beetroot, ajwa dates, fig and raisin are classified as low polyamines fruits while jujube demonstrates high content of polyamines. Significant decreased of total intracellular polyamine content after 24 h of treatments with these fruits were identified when compared with untreated A549 cells.Ornithine decarboxylase (ODC) activity showed downregulation upon treatment with ajwa dates, jujube and raisin while SSAT activity displayed alterations in beetroot and jujube treated A549 cells. Thus, we concluded that beetroot, ajwa dates, fig and raisins are the promising candidates for PDD strategy for their low polyamines, ability to reduce A549 intracellular polyamines and modification on ODC and SSAT activities

Investigation of polyamines as chemopreventive agent in selected prophetic food plants in human colon cancer-derived metastatic sw620 cells

Introduction: Polyamines are vital in maintaining human health because they performed certain functions that are necessary for cell development. It can be obtained by endogenous synthesis or exogenous supply through diet. Increasing intracellular polyamine content however promotes unwarranted cell proliferation and might as well stimulate cancer initiation. Thus, it is crucial to evaluate polyamine content in herbs that claimed to cure cancer to avoid unnecessary cell growth and proliferation. Therefore, this study aimed to investigate polyamines as chemopreventive agent using selected prophetic medicinal plants on human lung adenocarcinoma cells, A549. Methods: Five prophetic fruits were selected included ajwa dates (Phoenix dactylifera), beetroot (Beta vulgaris), fig (Ficus auriculata), jujube (Ziziphus jujube) and raisin (Vitis vinifera). These fruits were freeze dried with freeze-drying machine and stored in -80°C. Polyamine concentration in each fruit was quantified by High Performance Liquid Chromatography (HPLC). Subsequently, anti-proliferative effect of the fruits was evaluated in A549 cells using MTT assay and IC50 was determined. Later, growth effect of A549 cells was analyzed via Trypan blue exclusion assay and cell number and cell viability were recorded. Cells fraction from growth effect study was then used to measure protein content and intracellular polyamine concentration. From this, correlation between cell number and protein content was obtained. Cell cycle profile and apoptosis assay were analyzed with flow cytometer while caspase assay was done using microtiter plate reader. Apoptosis and caspase assay allowed type and mechanism of cell death to be detemined. Finally, expression of ornithine decarboxylase (ODC) and spermine/spermidine (N1)-acetyltransferase SSAT gene that regulate polyamine metabolic pathway was elucidated using qPCR. Results: Polyamine concentration in selected fruits showed significant highest polyamine concentration in jujube (219.6 ±4.4 nmoles/ml) while lowest in fig (39.3±3.0 nmoles/ml). MTT assay suggested IC50 was ranged from 15 mg/ml to 30 mg/ml. Growth effect analysis indicated significant decreased in cell number and cell viability after 24 hour of treatments. Protein content was decreased significantly and total elimination of intracellular spermidine and spermine was observed in treated A549 cells. There was correlation between cell number and protein content. Cell cycle profile displayed cell cycle arrest at G0/G1 after 48 hour of treatment in all treated cells. Annexin-v results demonstrated significant increase in early apoptosis in beetroot treated cells while other plants caused significant increase in late apoptosis at 48 hour of treatments. Caspase assay revealed significant activation of caspase 8 in cells treated with jujube and raisins while both caspase 8 and caspase 9 were activated in cells treated with ajwa dates, beetroot and fig. Expression of ODC gene and SSAT gene indicated significant downregulation of ODC gene in cells treated with ajwa and fig while upregulation of SSAT gene in beetroot, jujube and raisins treated cells. Conclusion: It was concluded that novel designs of nutritional therapeutic and preventive approaches for cancer using selected prophetic medicinal fruits are promising

The effect of Momordica Carantia (Bitter Mellon) extracts on human Adenocarcinoma cells proliferation and apoptosis

Abstract: Chemoprevention is a crucial process in overcoming the cancer burden throughout the world. By changing dietary choices, people who are at high risk of developing cancer may slow down the carcinogenesis or preventing it altogether. Some food contains high level of polyamine which is bad due to its ability for promoting tumor growth. Hence, this research aimed to determine the polyamine content of selected local plant, Momordica charantia by using High Performance Liquid Chromatographic (HPLC) analysis as well as conducting cytotoxicity study and growth inhibition study of the plant extracts against the human lung adenocarcinoma epithelial cell lines, A549. The type of cell death and its pathway also were elucidated. The polyamine content of M. charantia was found to be high (more than 200 nmol/g/mol). It is found to induce cytotoxicity against A549 cells after only 24 hours of exposure which lead to the establishment of the IC50 at 10.0 mg/ml. Growth curve analysis demonstrated that M. charantia had successfully reduced the cancer cell progression. On the other hand, there was an increase of intracellular polyamines in the treated cells compared to untreated cells. Apoptosis was induced Based on these preliminary findings, it can be concluded that M. charantia and G. procumbens are not an excellent plants to be included in the development of chemoprevention strategy as the ideal chemopreventive agents are those that contain low polyamine level, able to reduce the cancer cell progression and can inhibit the biosynthesis of polyamine in cancer cells to avoid recurrence of cancer. However, further investigation on the effect of these plants on polyamine metabolic pathway and cell death pathway shall be elucidated

Polyamines deficient diet is a complementary strategy in fighting cancer: an investigation on selected medicinal fruits

Polyamines are one of the oldest substances known in biochemistry consist of putrescine, spermidine and spermine. They are essential components of all living cells primarily involve in cell growth and proliferation. Mammalian cells are able to synthesize polyamines but are predominantly acquire them from exogenous sources such as intestinal flora and diets. However, the dietary sources provide a larger quantity of polyamines than the endogenous biosynthesis. Polyamines are tightly regulated by systematic biosynthesis and catabolism. Dysregulation of this system is highly associated with various diseases, including cancer. Therefore, polyamines deficient has become one of the strategies to inhibit carcinogenesis by targeting each biosynthetic and catabolic step in polyamines metabolism. Polyamine deficient diet (PDD) is a part of nutritional cancer therapy that recommends foods with low polyamines intake by cancer survivor and patients. In some cases, the plants might show anticancer effect but since the polyamines is high, it should not recommend for patients to avoid recurrence of cancer. Thus, this study was aimed to determine the polyamines content in selected anticancer fruits and plants in Malaysia and the effect on polyamine biosynthetic and catabolic enzymes was explored using human lung carcinoma cells (A549) as in vitro model. Methods: In this study, the selected fruits and plants are Phoenix dactylifera (ajwa dates), Beta vulgaris (beetroot), Ziziphus jujube (jujube), Vitis vinifera (raisin) and Punica granatum (pomegranate). The selected fruits and plants were initially grouped into classes based on total polyamines content using High performance Liquid Chromatography (HPLC). The ability of selected fruits to reduce A549 intracellular polyamines was quantified using the same technique. Accordingly, changes in polyamine metabolic enzymes activities; ornithine decarboxylase (ODC) and spermidine/spermine N-acetyltransferase (SSAT) were measured correspondingly using quantitative polymerase chain reactions (qPCR). Results: Results suggested polyamines content of selected fruits were ranging from low to high polyamines . Beetroot, ajwa dates, pomegranate and raisin are classified as low polyamines fruits while jujube demonstrate high content of polyamines (219.6 ± 4.43) nmol/mg. Significant decreased of total intracellular polyamine content after 24 h of treatments with selected fruits were identified when compared with untreated A549 cells (p<0.001). Ornithine decarboxylase activity showed changes upon treatment with P. dactylifera (ajwa dates), Z. jujube (jujube) and V. vinifera (raisin) while SSAT activity displayed alterations in B. vulgaris (beetroot) and Z. jujube (jujube) treated cells. Thus, we concluded that B. vulgaris (beetroot), P. dactylifera (ajwa dates) and V. vinifera (raisins) are the promising candidates for food-based chemoprevention strategy for their low polyamines and ability to reduce A549 intracellular polyamines. Conclusion and Significant of Study: The studies above strengthened the potential of polyamine reduced diet in cancer prevention. The polyamine deficient diet strategy is being documented as complementary step in chemoprevention in other countries. In United Kingdom, USA, Japan and Sweden, the database and investigation on local food and herbs has been established and being practiced. It is hope that this finding is beneficial for cancer patient, cancer survivor and even high risk of population to get cancer via genetic alteration and environmental exposure. We have investigated several local plants in Malaysia, however for this report, we focus on selected medicinal fruits as recommended by Rasulullah SAW

Ornithine decarboxylase gene expression in human lung adenocarcinoma cell (A549) treated with pomegranate juice

The polyamine biosynthesis pathway plays a significant role in cell growth, both normal and malignant. As polyamines are crucial in cellular growth and differentiation, they are linked to the development of cancer, with higher polyamine level observed in cancerous cells than in healthy cells. Accordingly, suppressing the polyamine pathway has been found to disrupt tumour development. Chemoprevention is considered a more feasible option in cancer management than chemotherapy, with a focus on natural chemopreventive agent. Pomegranate is known to inhibit several progression of lung cancer, although prior studies on the chemopreventive effect of pomegranate on lung cancer did not explore into polyamine pathway. Hence, this study investigated the effect of pomegranate juice on the polyamine pathway, by focusing on the biosynthesis involving ornithine decarboxylase (ODC), the rate limiting enzyme in the pathway. Quantitative polymerase chain reaction (qPCR) was applied to quantify the changes in ODC gene expression in A549 cells treated with pomegranate. The inhibition of growth was determined using Trypan Blue exclusion and the changes in intracellular polyamine in pomegranate treated cells was observed using High Performance Liquid Chromatography (HPLC). It was found that there was inhibition of A549 cell growth and reduced in intracellular polyamine content in pomegranate treated cells. The ODC expression was significantly inhibited compared to untreated cells, with a 48-fold difference. While this finding supports the hypothesis, there is much yet to be elucidated regarding its exact mechanism

Antiproliferative activity of selected prophetic medicinal plants on human lung adenocarcinoma cells, A549

Introduction: Cancer incidence has shown marked increase over the years that lead to significant interest in disease prevention strategies. Chemoprevention is one of the potential approaches using natural, synthetic or biological agents that are able to delay, reverse, or prevent cancer initiation. Using natural agents of recommended prophetic medicinal plants is our main focus owing to not much study has elaborated on them. Therefore, the aim of the present study is to investigate the effect of selected prophetic medicinal plants as potential cancer chemopreventive agents using fig (Ficus auriculata), dates (Phoenix dactylifera), and raisin (Vitis vinifera) against A549 cells. Methods: The plants were freeze dried with freeze-drying machine, diluted to a certain concentration and tested for antiproliferative activity using MTT and trypan blue (TB) exclusion assay. IC50, cell number and cell viability (%) were determined. Results: All plants showed significant dose-response relationship against A549. Fig and dates inhibited 50% of the cell population at concentration of 25 mg/ml while raisin at 30 mg/ml respectively. Growth curve indicated cell number was reduced over time from 0 to 144 hour and cell viability was ranged from 98.4-0%. Conclusion: The results suggested that selected prophetic medicinal plants tested displayed growth inhibitory effect in A549 cells