Polyamine is a group of aliphatic amine that is necessary for cell growth and development. However, potential of developing cancer is enhanced with the increase of polyamine levels in the body. Since polyamines can be obtained through diet intake, it is essential for cancer patients to consume foods that have low level of polyamines. Therefore, this study aims to determine the polyamine level of selected plants; Sabah Snake Grass (Clinacanthus nutans), Daun Kadok (Piper sarmentosum) and Fig (Ficus auriculata). The antiproliferative effects of C. nutans, P. sarmentosum and F. auriculata were investigated using MTT assay and the polyamines content in these plants and intracellular content were quantified using High Performance Liquid Chromatography (HPLC). The results showed that polyamines content in C. nutans (41.49 nmol/g), P. sarmentosum (55.01 nmol/g) anf F. auriculata (39.28 nmol/g) were classified under low level. The IC50 values for these plants were in the range of 20-30 mg/ml. This study demonstrated that these plants inhibit the A549 cell’s growth after 24 hours to 96 hours of exposure. Depletion of intracellular polyamine after 48 hours to 96 hours of exposure was also identified. In conclusion, the study showed that C. nutans, P. sarmentosum and F.auriculata contain low level of polyamine and have anti-proliferative effect against A549 cells. Therefore these plants are recommended to reduce or delayed the occurrence of malignancy and prevention of cancer recurrence after successful treatment.
Lung cancer is the most common type of cancer which the mortality rate increases year by year. Therapeutic drugs could not control the progression of cancer and it contributes to the side effects in normal cells. Thus, an alternative strategy using natural product becomes a focus today. Punica granatum, known as pomegranate has demonstrated the anti-proliferative effect in A549 cells. To further confirm its efficacy, this study aimed to investigate the type of cell death and its pathway in A549 cells. Propium Iodide staining was applied to determine the cell cycle profile changes induced by this juice. The determination of type of cell death was done using Annexin-V staining and later will be analysed using flowcytometer. The pathway to apoptosis was investigated by determining the caspase- 3, 8 and 9 activities. The findings were supported by mitochondrial membrane permeability assay and cytochrome c release detection which were later analysed using flowcytometer. This study revealed that pomegranate juice induced cell cycle arrest at G0/G1 phase and apoptosis through intrinsic pathway following 24 h treatment. Pomegranate juice caused loss of mitochondrial membrane permeability after 48 h [p<0.05] exposure and a release of cytochrome c in cytosol after 24 h [p<0.05] and 48 h [p<0.01] exposure in treated A549 cells. In caspases analysis, it was showed that there was activation of caspase-3 following 72 h [p<0.01] treatment and caspase-9 after 48 [p<0.01] and 72 h [p<0.05] exposure in treated A549 cells. It can be concluded that pomegranate juice able to cause A549 cell growth inhibition by inducing cell cycle arrest and apoptosis through mitochondrial pathway.
Lung cancer is the most common type of cancer which the mortality rate increases year by year. Therapeutic drugs could not control the progression of cancer and it contributes to the side effects in normal cells. Thus, an alternative strategy using natural product becomes a focus today. Punica granatum, known as pomegranate has demonstrated the anti-proliferative effect in A549 cells. To further confirm its efficacy, this study aimed to investigate the type of cell death and its pathway in A549 cells. Propium Iodide staining was applied to determine the cell cycle profile changes induced by this juice. The determination of type of cell death was done using Annex in-V staining and later will be analyzed using flow cytometer. The pathway to apoptosis was investigated by determining the caspase- 3, 8 and 9 activities. The findings were supported by mitochondrial membrane permeability assay and cytochrome c release detection which were later analyzed using flow cytometer. This study revealed that pomegranate juice induced cell cycle arrest at G0/G1 phase and apoptosis through intrinsic pathway following 24 h treatment. Pomegranate juice caused loss of mitochondrial membrane permeability after 48 h (p<0.05) exposure and a release of cytochrome c in cytosol after 24 h (p<0.05) and 48 h (p<0.01) exposure in treated A549 cells. In caspases analysis, it was showed that there was activation of caspase-3 following 72 h (p<0.01) treatment and caspase-9 after 48 (p<0.01) and 72 h (p<0.05) exposure in treated A549 cells. It can be concluded that pomegranate juice able to cause A549 cell growth inhibition by inducing cell cycle arrest and apoptosis through mitochondrial pathway.
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.
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