scholarly journals Piper sarmentosum induced apoptosis via mitochondrial pathway in human lung adenocarcinoma cells, A549

2016 ◽  
Vol 15 (1) ◽  
Author(s):  
Azila Sirajudeen ◽  
Aisyah Hanani Mohd Tahir ◽  
Radiah Abdul Ghani
Keyword(s):  
Lung Cancer ◽  
Cell Cycle ◽  
Cell Death ◽  
Cytochrome C ◽  
Treatment Strategies ◽  
A549 Cells ◽  
Annexin V ◽  
Mitochondrial Pathway ◽  
Cell Cycle Profile ◽  
Induced Apoptosis

Introduction: Lung cancer has been reported as one of the most common types of cancer worldwide. Current cancer treatments like chemotherapy do not result in a complete cure and are known to cause side effects in the patients. Therefore, alternative treatment strategies are being explored, one of which is to investigate the potential of the local herbs in this regard. Piper sarmentosum (daun kaduk) has received much attention due to its anti-cancer properties in A549 cells. In this study, the cell cycle profile and mechanisms of cell death induced by P. sarmentosum were investigated using a flow cytometer. Methods: The cell cycle profile changes were observed using propidium iodide staining while the type of cell death was analyzed using Annexin-V assay. Caspases -3/7,8 and 9 and cytochrome c assays were elucidated using flow cytometry analysis. Results: P.sarmentosum arrested the growth of A549 cells at G0/G1 phase. The Annexin V analysis revealed that P. sarmentosum exhibited significant induction of apoptosis after 24 h exposure. Caspases analysis showed that P. sarmentosum induced apoptosis through mitochondrial pathway, via the activation of caspase 3 and caspase 9. Meanwhile, cytochrome c analysis revealed that P. sarmentosum induced a mitochondrial pathway of cell death through the release of cytochrome c. Conclusions: Based on these preliminary findings, P. sarmentosum has a great potential as a dietary cancer treatment for lung cancer and may perhaps be used for lung cancer pharmacotherapy in the clinical settings in future.

MG132 as a proteasome inhibitor induces cell growth inhibition and cell death in A549 lung cancer cells via influencing reactive oxygen species and GSH level

2010 ◽  
Vol 29 (7) ◽  
pp. 607-614 ◽  
Author(s):  
Yong Hwan Han ◽  
Woo Hyun Park
Keyword(s):  
Lung Cancer ◽  
Reactive Oxygen Species ◽  
Cell Cycle ◽  
Cell Death ◽  
Cancer Cells ◽  
Proteasome Inhibitor ◽  
A549 Cells ◽  
Lung Cancer Cells ◽  
Oxygen Species ◽  
A549 Lung

Carbobenzoxy-Leu-Leu-leucinal (MG132) as a proteasome inhibitor has been shown to induce apoptotic cell death through formation of reactive oxygen species (ROS). In the present study, we evaluated the effects of MG132 on the growth of A549 lung cancer cells in relation to cell growth, ROS and glutathione (GSH) levels. Treatment with MG132 inhibited the growth of A549 cells with an IC50 of approximately 20 μM at 24 hours. DNA flow cytometric analysis indicated that 0.5 ∼ 30 μM MG132 induced a G1 phase arrest of the cell cycle in A549 cells. Treatment with 10 or 30 μM MG132 also induced apoptosis, as evidenced by sub-G1 cells and annexin V staining cells. This was accompanied by the loss of mitochondrial membrane potential (MMP; Δψm). The intracellular ROS levels including O2•- were strongly increased in 10 or 30 μM MG132-treated A549 cells but were down-regulated in 0.1, 0.5 or 1 μM MG132-treated cells. Furthermore, 10 or 30 μM MG132 increased mitochondrial O2•- level but 0.1, 0.5 or 1 μM MG132 decreased that. In addition, 10 or 30 μM MG132 induced GSH depletion in A549 cells. In conclusion, MG132 inhibited the growth of human A549 cells via inducing the cell cycle arrest as well as triggering apoptosis, which was in part correlated with the changes of ROS and GSH levels. Our present data provide important information on the anti-growth mechanisms of MG132 in A549 lung cancer cells in relation to ROS and GSH.

scholarly journals Tetrazolium Violet Induced Apoptosis and Cell Cycle Arrest in Human Lung Cancer A549 Cells

2012 ◽  
Vol 20 (2) ◽  
pp. 177-182 ◽  
Author(s):  
Xiao-Hong Zhang ◽  
Nan Zhang ◽  
Jian-Mei Lu ◽  
Qing-Zhong Kong ◽  
Yun-Feng Zhao
Keyword(s):  
Lung Cancer ◽  
Cell Cycle ◽  
Cell Cycle Arrest ◽  
Human Lung ◽  
A549 Cells ◽  
Human Lung Cancer ◽  
Cycle Arrest ◽  
Tetrazolium Violet ◽  
Induced Apoptosis

scholarly journals Pomegranate Juice Induced Cell Cycle Arrest and Apoptosis Via Mitochondrial Pathway in Human Lung Aden carcinoma A549 Cells

2018 ◽  
Vol 7 (3.21) ◽  
pp. 287
Author(s):  
Radiah Abdul Ghani ◽  
Nik Nurasyikin Nik Abdul Malek ◽  
Noor Suryani Mohd Ashari ◽  
Norzamzila Abdullah
Keyword(s):  
Cell Cycle ◽  
Cell Death ◽  
Cell Cycle Arrest ◽  
Mitochondrial Membrane ◽  
Caspase 3 ◽  
A549 Cells ◽  
Mitochondrial Pathway ◽  
Pomegranate Juice ◽  
Mitochondrial Membrane Permeability ◽  
Cycle Arrest

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 extract, LXB-1, from the barks of Liriodendron × hybrid, induced apoptosis via Akt, JNK and ERK1/2 pathways in A549 lung cancer cells

2015 ◽  
Vol 70 (11-12) ◽  
pp. 305-311 ◽  
Author(s):  
Jin-Hui Chen ◽  
Sen-Sen Lin ◽  
Wei-Xin Wang ◽  
Sheng-Tao Yuan ◽  
Ji-Sen Shi ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Western Blot ◽  
A549 Cells ◽  
Annexin V ◽  
Caspase 9 ◽  
M Cell ◽  
Protein Levels ◽  
Cycle Arrest ◽  
Induced Apoptosis ◽  
Human Lung Adenocarcinoma Cell

Abstract The effect of LXB-1, an extract from Liriodendron × hybrid, was determined on A549 human lung adenocarcinoma cell lines. Growth inhibition of LXB-1 was analyzed by MTT assay. Cancer cell cycle was measured by flow cytometry. To verify the apoptosis effect of LXB-1 on A549 cells, annexin V/PI double staining assay was performed. The expression levels of proapoptotic proteins were also measured by western blot. The potential mechanisms of LXB-1 inducing apoptosis – the expression and phosphorylation of ERK, p38, JNK and Akt – were investigated by western blot. The IC50 values of LXB-1 on A549 for 24, 48 and 72 h treatment were determined to be 12.97±1.53 μg/mL, 9.55±1.42 μg/mL, and 5.90±0.74 μg/mL, respectively. LXB-1 induced an obvious G2/M cell cycle arrest in A549 cells and resulted in significant cell apoptosis. LXB-1 also increased the cleavage of both caspase-3 and caspase-9, and greatly decreased the protein levels of Bcl-2. Moreover, LXB-1 increased the expression of phosphorylated JNK but decreased the levels of phosphorylated ERK1/2 and Akt. These results suggest that that LXB-1 induced apoptosis through JNK, ERK1/2, and Akt pathways in A549 cells.

scholarly journals Apoptosis Effect of Girinimbine Isolated fromMurraya koenigiion Lung Cancer CellsIn Vitro

2013 ◽  
Vol 2013 ◽  
pp. 1-12 ◽  
Author(s):  
Syam Mohan ◽  
Siddig Ibrahim Abdelwahab ◽  
Shiau-Chuen Cheah ◽  
Mohd Aspollah Sukari ◽  
Suvitha Syam ◽  
...  
Keyword(s):  
Lung Cancer ◽  
Cell Death ◽  
Membrane Potential ◽  
Cytochrome C ◽  
Mitochondrial Membrane Potential ◽  
Mitochondrial Membrane ◽  
Annexin V ◽  
Murraya Koenigii ◽  
Plasma Membrane Permeability ◽  
Lysosomal Stability

Murraya koenigiiSpreng has been traditionally claimed as a remedy for cancer. The current study investigated the anticancer effects of girinimbine, a carbazole alkaloid isolated fromMurraya koenigiiSpreng, on A549 lung cancer cells in relation to apoptotic mechanistic pathway. Girinimbine was isolated fromMurraya koenigiiSpreng. The antiproliferative activity was assayed using MTT and the apoptosis detection was done by annexin V and lysosomal stability assays. Multiparameter cytotoxicity assays were performed to investigate the change in mitochondrial membrane potential and cytochrome c translocation. ROS, caspase, and human apoptosis proteome profiler assays were done to investigate the apoptotic mechanism of cell death. The MTT assay revealed that the girinimbine induces cell death with an IC50of 19.01 μM. A significant induction of early phase of apoptosis was shown by annexin V and lysosomal stability assays. After 24 h treatment with 19.01 μM of girinimbine, decrease in the nuclear area and increase in mitochondrial membrane potential and plasma membrane permeability were readily visible. Moreover the translocation of cytochrome c also was observed. Girinimbine mediates its antiproliferative and apoptotic effects through up- and downregulation of apoptotic and antiapoptotic proteins. There was a significant involvement of both intrinsic and extrinsic pathways. Moreover, the upregulation of p53 as well as the cell proliferation repressor proteins, p27 and p21, and the significant role of insulin/IGF-1 signaling were also identified. Moreover the caspases 3 and 8 were found to be significantly activated. Our results taken together indicated that girinimbine may be a potential agent for anticancer drug development.

scholarly journals PNU-74654 Suppresses TNFR1/IKB Alpha/p65 Signaling and Induces Cell Death in Testicular Cancer

2022 ◽  
Vol 44 (1) ◽  
pp. 222-232
Author(s):  
Wen-Jung Chen ◽  
Wen-Wei Sung ◽  
Chia-Ying Yu ◽  
Yu-Ze Luan ◽  
Ya-Chuan Chang ◽  
...  
Keyword(s):  
Cell Death ◽  
Testicular Cancer ◽  
Cell Viability ◽  
Cell Lines ◽  
Cancer Progression ◽  
Treatment Strategies ◽  
Annexin V ◽  
Western Blots ◽  
Execution Phase ◽  
Induced Apoptosis

Testicular cancer (TC) is a rare malignancy worldwide and is the most common malignancy in males aged 15–44 years. The Wnt/β-catenin signaling pathway mediates numerous essential cellular functions and has potentially important effects on tumorigenesis and cancer progression. The search for drugs to inhibit this pathway has identified a small molecule, PNU-74654, as an inhibitor of the β-catenin/TCF4 interaction. We evaluated the therapeutic role of PNU-74654 in two TC cell lines, NCCIT and NTERA2, by measuring cell viability, cell cycle transition and cell death. Potential pathways were evaluated by protein arrays and Western blots. PNU-74654 decreased cell viability and induced apoptosis of TC cells, with significant increases in the sub G1, Hoechst-stained, Annexin V-PI-positive rates. PNU-74654 treatment of both TC cell lines inhibited the TNFR1/IKB alpha/p65 pathway and the execution phase of apoptosis. Our findings demonstrate that PNU-74654 can induce apoptosis in TC cells through mechanisms involving the execution phase of apoptosis and inhibition of TNFR1/IKB alpha/p65 signaling. Therefore, small molecules such as PNU-74654 may identify potential new treatment strategies for TC.

Evaluation of Cytotoxic Mechanisms of Clinacanthus Nutans Extracts in Cancer Cells

2020 ◽  
Vol 10 ◽  
Author(s):  
Nurul Atiqah Sulaiman ◽  
Rajan Rajabalaya ◽  
Shirley Huan Fang Lee ◽  
Ya Chee Lim ◽  
Wei Hon Lim ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Cell Cycle ◽  
Flow Cytometry ◽  
Cell Death ◽  
Cancer Cells ◽  
Cancer Cell ◽  
Annexin V ◽  
Cell Cycle Profile ◽  
Clinacanthus Nutans ◽  
Mcf 7

Background: Commercially available Clinacanthus nutans (Burm.F) Lindau (Acanthaceae) (CN) leaf preparations are gaining attention as an alternative cancer treatment, particularly in South East Asia. Multiple studies have suggested that CN has potential anticancer activities; however, the mechanism of these activities has remained elusive. Objective: This study evaluated the cytotoxic mechanisms of CN extracts in cancer cells. Methods: CN extracts were prepared from either fresh or dried leaves, using different solvents. Cytotoxicity of CN extracts were tested on the A549 (lung cancer), HeLa (cervical cancer), MCF-7 (breast cancer) and MDA-MB-231 (breast cancer) cell lines using the MTT assay. Flow cytometry was used to assess changes in the cell cycle profile, while Western blotting was used to examine microtubule stability. Finally, the mode of cell death was investigated using the Annexin V-FITC Apoptosis Detection Kit. Results: Aqueous Fresh (AQF) extract was prepared to simulate the ethno-medicinal use of CN, and reduced cell viability of MCF-7 cells with IC50 = 1.71 mg/mL. Some CN extracts have the ability to inhibit the proliferation of four different cancer cell lines after a 24 hour treatment. Annexin V assay results shows that acetone extracts of CN induced increments in percentage of apoptotic cell death. However, flow cytometry results show that cancer cell cycle profile were not affected. Similarly, immunoblotting results also indicate that microtubule dynamics in MCF-7 cells were not altered. However, the aqueous extract, prepared to simulate the current ethnomedicinal use of CN leaves in cancer treatment, did not significantly inhibit cancer cell proliferation with IC50 = 1.71 mg/mL. Conclusions: This study was the first to show that microtubules in cancer cells remain dynamic after treatment with CN extracts, effectively ruling interference of microtubule dynamics as the mode of cell death. AMD extract showed the highest effects MCF-7 cell proliferation.

scholarly journals The role of ARK in stress-induced apoptosis in Drosophila cells

2002 ◽  
Vol 156 (6) ◽  
pp. 1077-1087 ◽  
Author(s):  
Katja C. Zimmermann ◽  
Jean-Ehrland Ricci ◽  
Nathalie M. Droin ◽  
Douglas R. Green
Keyword(s):  
Cell Death ◽  
Cytochrome C ◽  
Molecular Mechanisms ◽  
Mitochondrial Pathway ◽  
Exact Function ◽  
Increased Sensitivity ◽  
Induced Apoptosis ◽  
Drosophila Cells ◽  
Pronounced Inhibition

The molecular mechanisms of apoptosis are highly conserved throughout evolution. The homologs of genes essential for apoptosis in Caenorhabditis elegans and Drosophila melanogaster have been shown to be important for apoptosis in mammalian systems. Although a homologue for CED-4/apoptotic protease-activating factor (Apaf)-1 has been described in Drosophila, its exact function and the role of the mitochondrial pathway in its activation remain unclear. Here, we used the technique of RNA interference to dissect apoptotic signaling pathways in Drosophila cells. Inhibition of the Drosophila CED-4/Apaf-1–related killer (ARK) homologue resulted in pronounced inhibition of stress-induced apoptosis, whereas loss of ARK did not protect the cells from Reaper- or Grim-induced cell death. Reduction of DIAP1 induced rapid apoptosis in these cells, whereas the inhibition of DIAP2 expression did not but resulted in increased sensitivity to stress-induced apoptosis; apoptosis in both cases was prevented by inhibition of ARK expression. Cells in which cytochrome c expression was decreased underwent apoptosis induced by stress stimuli, Reaper or Grim. These results demonstrate the central role of ARK in stress-induced apoptosis, which appears to act independently of cytochrome c. Apoptosis induced by Reaper or Grim can proceed via a distinct pathway, independent of ARK.

scholarly journals Effects of Holothurian Glycosaminoglycan on the Sensitivity of Lung Cancer to Chemotherapy

2020 ◽  
Vol 19 ◽  
pp. 153473542091143
Author(s):  
Cunzhi Lin ◽  
Xinhong Zhu ◽  
Qing Jin ◽  
Aihua Sui ◽  
Jinfeng Li ◽  
...  
Keyword(s):  
Lung Cancer ◽  
Cell Cycle ◽  
Sea Cucumber ◽  
Caspase 3 ◽  
A549 Cells ◽  
Annexin V ◽  
Inhibitory Effect ◽  
The Body ◽  
Control Group ◽  
Apoptosis Pathway

Sea cucumber is a kind of food. Holothurian glycosaminoglycan (hGAG) is extracted from the body wall of the sea cucumber. Administration of hGAG and cisplatin (DDP) together to treat lung cancer was investigated. Lung adenocarcinoma A549 cells were cultured and divided into 4 groups: control group, hGAG 100 µg/mL group, DDP 3 µg/mL group, and hGAG 100 µg/mL + DDP 3 µg/mL group. Cell inhibition and apoptosis was evaluated by CCK8 and Hoechst33258 staining. Cell cycle was tested by Annexin V-FITC/PI (propidium iodide) double-staining and flow cytometry. The expression of mRNA and protein of Bcl-2, Bax, caspase-3, and survivin were detected by reverse transcriptase-polymerase chain reaction and Western blot, respectively. The results showed that hGAG combined with DDP enhanced the inhibitory effect of DDP on A549 lung cells through apoptosis pathway. The mechanism of apoptosis may be related to the reduction of Bcl-2 and survivin, as well as the ascension of Bax and caspase-3. hGAG could promote A549 cell cycle arrest in G1 and G2 phase and improve the DDP chemotherapy effects on A549 cells.

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