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

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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Inhibitory effect of lignans from Ailanthus altissima on proliferation of human lung cancer cells and its mechanism.

Journal of Clinical Oncology ◽

10.1200/jco.2021.39.15_suppl.e15059 ◽

2021 ◽

Vol 39 (15_suppl) ◽

pp. e15059-e15059

Author(s):

Yu Wang ◽

Siming Wang ◽

Yibing Wu ◽

Mei Dong ◽

Zhiyu Ni

Keyword(s):

Lung Cancer ◽

Caspase 3 ◽

Human Lung ◽

A549 Cells ◽

Inhibitory Effect ◽

Human Lung Cancer ◽

Ailanthus Altissima ◽

Control Group ◽

Caspase Inhibitor ◽

Significant Difference

e15059 Background: Plants are thought to contain compounds that inhibit the proliferation of cancer cells in vitro, and many attempts have been made to isolate anti-cancer drugs from plants. Ailanthus altissima is a family of Simaroubaceae, the bark is gray to grayish-black, because of the smell of the gland patches at the base of the leaves. 3,5,3-trimethoxy-8,1-neoligna-4,2,7,9,9-pentol (TNP) isolated from Ailanthus altissima. Methods: (1) Apoptosis was detected by flow cytometry: the cells were incubated with 10 mmol/L TNP for 24 h, and the cells were collected. The cell concentration was adjusted to 5×105 ̃ 5×106/ L. While propionate iodide was added for DNA fluorescence staining (Propidium Iodide, PI: 50 mg/L，triton-x100 1.0%). (2) The changes of p53, Bax and caspase-3 were detected by Western blot. The cells were cultured in 10 μmol/L cisplatin and TNP medium for 24 hours. Image J image analysis software is used for semi quantitative analysis. The data were expressed by mean standard deviation, and the mean values of each group were compared by analysis of variance and the least significant difference. There was a significant difference in P< 0.05. (3) A549 cells were treated with 10 μmol/L cisplatin and TNP. A549 cells were incubated with cisplatin and TNP, and then treated with solvent control (final concentration 0.1% dimethyl sulfoxide) with or without caspase inhibitor for 48 hours The OD value was determined by MTT method. Cell survival rate (%) = (OD value of control group /OD value of control group) 100%. Results: (1) The reverse effect of caspase inhibitor on the proliferation of A549 cells: the cell survival rates of A549 cells treated with 0.1, 1 and 10 μmol/L cisplatin and TNP were 96.11%, 72.36%, 27.57% and 71.42%, 28.97% and 4.34% respectively. The survival rate of A549 cells incubated with cisplatin and TNP 20 μmol/L increased to 100.00%, 81.92%, 57.54% and 85.69%, 48.57% and 16.95% respectively. Caspase inhibitor reversed the inhibitory effect of TNP on A549 cells. (2) The effect of TNP on apoptosis of A549 cells: after treatment with 10 μmol/L TNP for 24 h, the apoptosis rate of A549 cells was (24.01)%, which was significantly higher than that of the blank control group (6.79)% ( P< 0.05). (3) The results showed that the up regulation of p53, Bax and caspase-3 protein by TNP was 10 μmol/L. Conclusions: TNP can induce apoptosis of human lung cancer cell line A549, and it can up regulate the expression of p53, Bax and Caspase-3, which may be one of the mechanisms of TNP inducing apoptosis of A549 cells. The experiment also proves that caspase inhibitor can reverse the effect of TNP. TNP inhibits the proliferation of A549 cells.

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The pro-apoptosis effects of Echinacea purpurea and Cannabis sativa extracts in human lung cancer cells through caspase-dependent pathway

BMC Complementary Medicine and Therapies ◽

10.1186/s12906-021-03204-6 ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

Fatemeh Hosami ◽

Azadeh Manayi ◽

Vahid Salimi ◽

Farshad Khodakhah ◽

Mitra Nourbakhsh ◽

...

Keyword(s):

Lung Cancer ◽

Cell Cycle ◽

Caspase 3 ◽

Cannabis Sativa ◽

A549 Cells ◽

Echinacea Purpurea ◽

G1 Phase ◽

Cell Cycle Distribution ◽

Cb2 Receptor ◽

Anti Cancer

Abstract Background Considering the advantages of using medicinal herbs as supplementary treatments to sensitize conventional anti-cancer drugs, studying functional mechanisms and regulatory effects of Echinacea purpurea (as a non-cannabinoid plant) and Cannabis sativa (as a cannabinoid plant) are timely and required. The potential effects of such herbs on lung cancer cell growth, apoptosis, cell cycle distribution, cellular reactive oxygen species (ROS) level, caspase activity and their cannabinomimetic properties on the CB2 receptor are addressed in the current study. Methods The cytotoxic effect of both herb extracts on the growth of lung cancer cells (A549) was assessed using the MTT assay. The annexin-V-FITC staining and propidium iodide (PI) staining methods were applied for the detection of apoptosis and cell cycle distribution using flow cytometry. The cellular level of ROS was measured using 7′-dichlorofluorescin diacetate (DCFH-DA) as a fluorescent probe in flow cytometry. The caspase 3 activity was assessed using a colorimetric assay Kit. Results Echinacea purpurea (EP) root extract induced a considerable decrease in A549 viable cells, showing a time and dose-dependent response. The cell toxicity of EP was accompanied by induction of early apoptosis and cell accumulation at the sub G1 phase of the cell cycle. The elevation of cellular ROS level and caspase 3 activity indicate ROS-induced caspase-dependent apoptosis following the treatment of A549 cells by EP extract. The observed effects of EP extract on A549 growth and death were abrogated following blockage of CB2 using AM630, a specific antagonist of the CB2 receptor. Increasing concentrations of Cannabis sativa (CS) induced A549 cell death in a time-dependent manner, followed by induction of early apoptosis, cell cycle arrest at sub G1 phase, elevation of ROS level, and activation of caspase 3. The CB2 blockage caused attenuation of CS effects on A549 cell death which revealed consistency with the effects of EP extract on A549 cells. Conclusions The pro-apoptotic effects of EP and CS extracts on A549 cells and their possible regulatory role of CB2 activity might be attributed to metabolites of both herbs. These effects deserve receiving more attention as alternative anti-cancer agents. Graphical abstract

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Effects of 5,6-Dihydroxy-2,4-Dimethoxy-9,10-Dihydrophenanthrene on G2/M Cell Cycle Arrest and Apoptosis in Human Lung Carcinoma Cells

The American Journal of Chinese Medicine ◽

10.1142/s0192415x16500828 ◽

2016 ◽

Vol 44 (07) ◽

pp. 1473-1490 ◽

Cited By ~ 6

Author(s):

Wipada Duangprompo ◽

Kalaya Aree ◽

Arunporn Itharat ◽

Pintusorn Hansakul

Keyword(s):

Lung Cancer ◽

Cell Cycle ◽

Cell Cycle Arrest ◽

Caspase 3 ◽

Human Lung ◽

A549 Cells ◽

Human Lung Cancer ◽

Apoptotic Cells ◽

Mrna Levels ◽

Cycle Arrest

5,6-dihydroxy-2,4-dimethoxy-9,10-dihydrophenanthrene (HMP) is an active compound isolated from the rhizome extracts of Dioscorea membranacea Pierre, a Thai medicinal plant. This study aimed to investigate the growth-inhibitory and apoptosis-inducing effects of HMP in human lung cancer A549 cells. The antiproliferative and cytotoxic effects of HMP were analyzed by a Sulforhodamine B assay. Cell division, cell cycle distribution and membrane asymmetry changes were each performed with different fluorescent dyes and then analyzed by flow cytometry. Real-time PCR and immunoblotting were used to detect cell cycle- and apoptosis-related mRNA levels and proteins, respectively. The nuclear morphology of the cells stained with DAPI and DNA fragmentation were detected by fluorescence microscopy and gel electrophoresis, respectively. The results showed that HMP exerted strong antiproliferative and cytotoxic activities in A549 cells with the highest selectivity index. It halted the cell cycle in [Formula: see text]/M phase via down-regulation of the expression levels of regulatory proteins Cdc25C, Cdk1 and cyclinB1. In addition, HMP induced early apoptotic cells with externalized phosphatidylserine and subsequent apoptotic cells in sub-[Formula: see text] phase. HMP increased caspase-3 activity and levels of the cleaved (active) form of caspase-3 whose actions were supported by the cleavage of its target PARP, nuclear condensation and DNA apoptotic ladder. Moreover, HMP significantly increased the mRNA and protein levels of proapoptotic Bax as well as promoted subsequent caspase-9 activation and BID cleavage, indicating HMP-induced apoptosis via both intrinsic and extrinsic pathways. These data support, for the first time, the potential role of HMP as a cell-cycle arrest and apoptosis-inducing agent for lung cancer treatment.

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

IIUM Medical Journal Malaysia ◽

10.31436/imjm.v15i1.1275 ◽

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.

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Apoptotic effect of astaxanthin from white shrimp shells on lung cancer A549 cells

Tropical Journal of Pharmaceutical Research ◽

10.4314/tjpr.v19i9.6 ◽

2020 ◽

Vol 19 (9) ◽

pp. 1835-1842

Author(s):

Supita Tanasawet ◽

Wanida Sukketsiri ◽

Pennapa Chonpathompikunlert ◽

Wanwimol Klaypradit ◽

...

Keyword(s):

Lung Cancer ◽

Cell Cycle ◽

Membrane Potential ◽

Mitochondrial Membrane Potential ◽

Litopenaeus Vannamei ◽

Mitochondrial Membrane ◽

Caspase 3 ◽

A549 Cells ◽

Nuclear Staining ◽

Dependent Manner

Purpose: To investigate the anti-cancer potential of astaxanthin from Litopenaeus vannamei encapsulated in liposomes (ASX) to treat lung cancer A549 cells.Methods: Lung adenocarcinoma A549 cells were cultured and treated with ASX, following which cell viability and nuclear staining were performed. Generation of ROS was identified by the DCFH-DA assay while tetramethylrhodamine ethyl ester was used to determine the mitochondrial membrane potential. Flow cytometry was applied to investigate caspase-3/7 activity and cell cycle distribution.Results: ASX inhibited growth of A549 in a concentration- and time- dependent manner. The IC50 values at 24, 48 and 72 h were 53.73, 22.85, 17.46 μg/mL, respectively (p < 0.05). After incubation with ASX, the morphological changes were observed in A549 cells following Hoechst 33342/PI fluorescent staining. ASX increased ROS generation and was associated with the collapse of mitochondrial membrane potential, which subsequently triggered the activation of caspase-3/7 activity leading to apoptosis (p < 0.05). In addition, A549 cells accumulated in the G0/G1 phase.Conclusion: The results suggest that ASX is a valuable nutraceutical agent to target A549 lung cancer cells via ROS-dependent pathway as well as blockage of cell cycle progression. Keywords: Astaxanthin, Litopenaeus vannamei, Lung cancer, A549, Apoptosis

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Forsythia suspensa extract has inhibitory effect on proliferation and apoptosis of A549 lung cancer cells

Tropical Journal of Pharmaceutical Research ◽

10.4314/tjpr.v18i9.24 ◽

2021 ◽

Vol 18 (9) ◽

pp. 1949-1954

Author(s):

Wei-guo Zhang ◽

Qin Liu ◽

Cai-peng Lei

Keyword(s):

Lung Cancer ◽

Cell Cycle ◽

Cancer Cells ◽

A549 Cells ◽

Inhibitory Effect ◽

Lung Cancer Cells ◽

Blue Staining ◽

G1 Phase ◽

Forsythia Suspensa ◽

A549 Lung

Purpose: To investigate the effect of Forsythia suspensa extract (FSE) on apoptosis and proliferation in A549 human lung cancer cells. Methods: Inverted microscope was employed to observe morphological changes in A549 cells after exposure to FSE. Trypan blue staining of living cells was used to construct the cell growth curve after treatment with varying concentrations of FSE. The influence of FSE on cell proliferation, apoptosis and cell cycle was determined by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay, while protein expressions of key apoptosis-related enzymes were evaluated by immunocytochemical method. Results: FSE inhibited the growth of A549 lung cancer cells at a concentration range of 10 - 150 μg/mL. Flow cytometry results showed that FSE induced apoptosis in A549 cells. The proportion of cells in G0/G1-phase increased significantly (p < 0.01), while the proportion of cells in S- and G2/M-phase decreased correspondingly, indicating that the cells were in G0/G1-phase arrest. Cell cycle arrest and apoptosis-inducing effect gradually rose with increase in FSE concentration. With increasing concentrations of FSE, there was also significant increase in the expressions of caspase-3 (p < 0.05), caspase-8 (p < 0.01) and caspase-9 (p < 0.05), but significant decrease in Ki-67 (p < 0.01) and p21 ras protein (p < 0.01). Conclusion: FSE exerts significant inhibitory effect on the proliferation of A549 lung cancer cells. Therefore, the plant can potentially be developed for the treatment of lung cancer.

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Jaceosidin Induces Apoptosis in U87 Glioblastoma Cells through G2/M Phase Arrest

Evidence-based Complementary and Alternative Medicine ◽

10.1155/2012/703034 ◽

2012 ◽

Vol 2012 ◽

pp. 1-12 ◽

Cited By ~ 25

Author(s):

Muhammad Khan ◽

Bo Yu ◽

Azhar Rasul ◽

Ali Al Shawi ◽

Fei Yi ◽

...

Keyword(s):

Cell Cycle ◽

Caspase 3 ◽

Ethanol Extract ◽

Inhibitory Effect ◽

Bioactive Constituents ◽

Growth Inhibitory Effect ◽

Apoptosis Pathway ◽

Growth Inhibitory ◽

M Phase ◽

Induction Of Apoptosis

Artemisia argyiis a widely used medicinal plant in China. The present study was designed to identify the bioactive constituents with antiglioma activity from leaves ofArtemesia argyi. A bioactivity guided approach based on MTT assay for cells growth inhibition led to the isolation of a flavonoid, “jaceosidin” from ethanol extract of leaves ofArtemesia argyi. The growth inhibitory effect of jaceosidin was explored using flow cytometry and Western blot studies. Our results showed that jaceosidin exerts growth inhibitory effect by arresting the cells at G2/M phase and induction of apoptosis. Furthermore, our study revealed that induction of apoptosis was associated with cell cycle arrest at G2/M phase, upregulation of p53 and Bax, decrease in mitochondrial membrane potential, release of cytochrome c, and activation of caspase 3. This mitochondrial-caspase-3-dependent apoptosis pathway was confirmed by pretreatment with caspase 3 inhibitor, Ac-DEVD-CHO. Our findings suggested that jaceosidin induces mitochondrial-caspase-3-dependent apoptosis in U87 cells by arresting the cell cycle at G2/M phase.

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Combined Treatment of Cinobufotalin and Gefitinib Exhibits Potent Efficacy against Lung Cancer

Evidence-based Complementary and Alternative Medicine ◽

10.1155/2021/6612365 ◽

2021 ◽

Vol 2021 ◽

pp. 1-9

Author(s):

Youxi Han ◽

Ronghui Ma ◽

Guolei Cao ◽

Hao Liu ◽

Lili He ◽

...

Keyword(s):

Lung Cancer ◽

Combined Therapy ◽

Combined Treatment ◽

A549 Cells ◽

Annexin V ◽

Inhibitory Effect ◽

Combination Group ◽

Tunel Staining ◽

Lung Cancer Patients ◽

Cycle Arrest

This study aimed to evaluate the efficacy of cinobufotalin combined with gefitinib in the treatment of lung cancer. A549 cells were treated with gefitinib, cinobufotalin, or cinobufotalin plus gefitinib. MTT assay, annexin-V/PI staining and flow cytometry, TUNEL staining, DCFH-DA staining, Western blot, and real-time RT-PCR were performed to investigate the synergistic inhibitory effect of cinobufotalin combined with gefitinib on the growth of A549 cells. Results showed that cinobufotalin synergized with gefitinib displayed inhibited cell viability and enhanced apoptosis in the combination group. Cinobufotalin combined with gefitinib induced a significant enhancement in reactive oxygen species (ROS) production accompanied by cell cycle arrest in the S phase arrest, characterized by upregulation of p21 and downregulation of cyclin A, cyclin E, and CDK2. Besides, cinobufotalin plus gefitinib downregulated the levels of HGF and c-Met. In summary, cinobufotalin combined with gefitinib impedes viability and facilitates apoptosis of A549 cells, indicating that the combined therapy might be a new promising treatment for lung cancer patients who are resistant to gefitinib.

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Screening of Apoptosis Pathway-Mediated Anti-Proliferative Activity of the Phytochemical Compound Furanodienone against Human Non-Small Lung Cancer A-549 Cells

Life ◽

10.3390/life12010114 ◽

2022 ◽

Vol 12 (1) ◽

pp. 114

Author(s):

Ahmed Al Saqr ◽

El-Sayed Khafagy ◽

Mohammed F. Aldawsari ◽

Khaled Almansour ◽

Amr S. Abu Lila

Keyword(s):

Lung Cancer ◽

Cell Cycle ◽

Cell Cycle Arrest ◽

Molecular Mechanisms ◽

Wnt Signaling Pathway ◽

A549 Cells ◽

Human Lung Cancer ◽

Dependent Manner ◽

Apoptosis Pathway ◽

Cycle Arrest

Furanodienone (FDN), a major bioactive component of sesquiterpenes produced from Rhizoma curcumae, has been repeatedly acknowledged for its intrinsic anticancer efficacy against different types of cancer. In this study, we aimed to investigate the cytotoxic potential of furanodienone against human lung cancer (NSCLC A549) cells in vitro, as well as its underlying molecular mechanisms in the induction of apoptosis. Herein, we found that FDN significantly inhibited the proliferation of A549 cells in a dose-dependent manner. In addition, treatment with FDN potentially triggered apoptosis in A549 cells via not only disrupting the nuclear morphology, but by activating capsase-9 and caspase-3 with concomitant modulation of the pro- and antiapoptotic gene expression as well. Furthermore, FDN revealed its competence in inducing cell cycle arrest at G0/G1 phase in A549 cells, which was associated with decreased expression of cyclin D1 and cyclin-dependent kinase 4 (CDK4), along with increased expression of CDK inhibitor p21Cip1. Intriguingly, FDN treatment efficiently downregulated the Wnt signaling pathway, which was correlated with increased apoptosis, as well as cell cycle arrest, in A549 cells. Collectively, FDN might represent a promising adjuvant therapy for the management of lung cancer.

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Effect and Mechanism of Transthyretin Over-Expression on Proliferation and Cell Cycle of Lung Cancer A549 Cells

Iranian Journal of Public Health ◽

10.18502/ijph.v50i4.5995 ◽

2021 ◽

Author(s):

Deqing Zhu ◽

Xuan Li ◽

Hao Gong ◽

Jing Li ◽

Xike Lu ◽

...

Keyword(s):

Lung Cancer ◽

Cell Cycle ◽

Cell Proliferation ◽

Protein Expression ◽

Mrna Expression ◽

Quantitative Polymerase Chain Reaction ◽

Statistical Significance ◽

A549 Cells ◽

Control Group ◽

Over Expression

Background: The effects of transthyretin (TTR) over-expression on the proliferation and cell cycle of nonsmall cell lung cancer (NSCLC) A549 cells and its possible mechanism were verified. Methods: A total of 196 LC patients and 20 healthy controls were enrolled at Tianjin Hospital, Tianjin, China between Apr 2017 and Oct 2017. The serum TTR content was detected by ELISA. Through lentiviral transfection method, NSCLC cells were divided into non-transfected group (group A), negative control group (group B) transfected with empty vector and experimental group (group C) transfected with TTR overexpression. Cell proliferation was detected by CCK-8 method, TTR mRNA expression was detected by realtime quantitative polymerase chain reaction (RT-qPCR), and TTR protein expression was tested by Western blot (WB). Cell cycle was detected by flow cytometry, Wnt3a/β-catenin protein expression was detected by WB, and mRNA expression was detected by RT-qPCR. Results: The serum TTR content in early, middle and late LC group was remarkably lower than that in healthy group (P<0.05). Compared with late stage, TTR content in early and middle stages of LC group was higher, and the difference was statistically marked (P < 0.05). The absorbance value of group C was lower than that of groups A and B, indicating that the cell proliferation activity dramatically decreased, with statistically marked difference (P<0.05). LC A549 cells in group C were obviously blocked in G2M, with statistical significance (P<0.05). Conclusion: TTR over-expression can inhibit the proliferation of NSCLC A549 cells, and the expression is related to Wnt3a/β-catenin pathway. TTR in serum of patients was helpful for diagnosing LC and has certain clinical value.

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