Kaempferide Induces G0/G1 Phase Arrest and Apoptosis via ROS-Mediated Signaling Pathways in A549 Human Lung Cancer Cells

Kaempferide is an O-methylated flavonol that has received much attention due to its various biological activities. In this study, we explored the underlying mechanisms of kaempferide in human lung cancer A549 cells. The Cell Counting Kit-8 (CCK-8) assay, Hoechst 33342/propidium iodide double staining, flow cytometry, scratch wound healing assay, and Western blot analysis were used to measure cell apoptosis, the cell cycle, reactive oxygen species (ROS) levels, and cell migration of human lung cancer cells. Kaempferide significantly inhibited human lung cancer cell proliferation, and its toxic effects on normal cells were significantly lower than those of 5-fluorouracil. Kaempferide induced A549 cell apoptosis by decreasing the mitochondrial membrane potential and the expression level of B-cell lymphoma 2, and by increasing the expression levels of Bcl-2-associated X protein and caspase-3. It also regulated mitogen-activated protein kinase (MAPK), signal transducer and activator of transcription 3 (STAT3), and nuclear factor kappa B (NF-κB) signaling pathways by increasing the expression levels of phosphorylated c-Jun N-terminal kinase, p-p38, I kappa B, and by decreasing the expression levels of phosphorylated extracellular signal-regulated kinase, p-STAT3, and NF-κB. Kaempferide induced cell cycle arrest in the G0/G1 phase in A549 cells by downregulating the expression levels of p-AKT, cyclin D1, and cyclin-dependent kinase 2. Furthermore, kaempferide blocked A549 cell migration by downregulating the expression levels of transforming growth factor beta 1 (TGF-β1), p-β-catenin, p-glycogen synthase kinase 3 beta, N-cadherin, and vimentin, and by upregulating the expression level of E-cadherin. Kaempferide enhanced the accumulation of ROS, and N-acetyl-l-cysteine (a ROS inhibitor) decreased the regulation of MAPK, NF-κB, AKT, and TGF-β signaling pathways by kaempferide, inhibited cell apoptosis, and reversed cell cycle arrest. Our results showed that kaempferide induced apoptosis via ROS-mediated MAPK, NF-κB, AKT, and TGF-β signaling pathways in A549 cells. Thus, kaempferide may be a novel drug candidate for lung cancer.

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10-HDA Induces ROS-Mediated Apoptosis in A549 Human Lung Cancer Cells by Regulating the MAPK, STAT3, NF-κB, and TGF-β1 Signaling Pathways

BioMed Research International ◽

10.1155/2020/3042636 ◽

2020 ◽

Vol 2020 ◽

pp. 1-15

Author(s):

Xin-Mei Lin ◽

Shao-Bin Liu ◽

Ying-Hua Luo ◽

Wan-Ting Xu ◽

Yu Zhang ◽

...

Keyword(s):

Lung Cancer ◽

Cell Cycle ◽

Cancer Cells ◽

Signaling Pathways ◽

Human Lung ◽

Lung Cancer Cells ◽

Human Lung Cancer ◽

Cycle Arrest ◽

Human Lung Cancer Cells ◽

Tgf Β1

10-Hydroxy-2-decenoic acid (10-HDA), also known as royal jelly acid, has a variety of physiological functions, and recent studies have shown that it also has anticancer effects. However, its anticancer mechanisms have not been clearly defined. In this study, we investigated the underlying mechanisms of 10-HDA in A549 human lung cancer cells. We used Cell Counting Kit-8 assay, scratch wound healing assay, flow cytometry, and western blot analysis to investigate its apoptotic effects and underlying mechanism. Our results showed that 10-HDA inhibited the proliferation of three types of human lung cancer cells and had no significant toxic effects on normal cells. Accompanying reactive oxygen species (ROS), 10-HDA induced A549 cell apoptosis by regulating mitochondrial-associated apoptosis, and caused cell cycle arrest at the G0/G1 phase in a time-dependent manner. Meanwhile, 10-HDA also regulated mitogen-activated protein kinase (MAPK), signal transducer and activator of transcription 3 (STAT3), and nuclear factor kappa B (NF-κB) signaling pathways by increasing the expression levels of phosphorylated c-Jun N-terminal kinase, p-p38, and I-κB, and additionally, by decreasing the expression levels of phosphorylated extracellular signal-regulated kinase, p-STAT3, and NF-κB. These effects were blocked by MAPK inhibitors and N-acetyl-L-cysteine. Furthermore, 10-HDA inhibited cell migration by regulating transforming growth factor beta 1 (TGF-β1), SNAI1, GSK-3β, E-cadherin, N-cadherin, and vimentin. Taken together, the results of this study showed that 10-HDA induced cell cycle arrest and apoptosis in A549 human lung cancer cells through ROS-mediated MAPK, STAT3, NF-κB, and TGF-β1 signaling pathways. Therefore, 10-HDA may be a potential therapy for human lung cancer.

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Isoorientin induces the apoptosis and cell cycle arrest of A549 human lung cancer cells via the ROS‑regulated MAPK, STAT3 and NF‑κB signaling pathways

International Journal of Oncology ◽

10.3892/ijo.2020.5079 ◽

2020 ◽

Vol 57 (2) ◽

pp. 550-561

Author(s):

Wan‑Ting Xu ◽

Gui‑Nan Shen ◽

Tian‑Zhu Li ◽

Yu Zhang ◽

Tong Zhang ◽

...

Keyword(s):

Lung Cancer ◽

Cell Cycle ◽

Cell Cycle Arrest ◽

Cancer Cells ◽

Signaling Pathways ◽

Human Lung ◽

Lung Cancer Cells ◽

Human Lung Cancer ◽

Cycle Arrest ◽

Human Lung Cancer Cells

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Overexpression of cyclin L2 induces apoptosis and cell-cycle arrest in human lung cancer cells

Chinese Medical Journal ◽

10.1097/00029330-200705020-00010 ◽

2007 ◽

Vol 120 (10) ◽

pp. 905-909 ◽

Cited By ~ 14

Author(s):

Hong-li LI ◽

Tong-shan WANG ◽

Xiao-yu LI ◽

Nan LI ◽

Ding-zhi HUANG ◽

...

Keyword(s):

Lung Cancer ◽

Cell Cycle ◽

Cell Cycle Arrest ◽

Cancer Cells ◽

Human Lung ◽

Lung Cancer Cells ◽

Human Lung Cancer ◽

Cycle Arrest ◽

Human Lung Cancer Cells

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Growth Inhibition and Induction of G1 Phase Cell Cycle Arrest in Human Lung Cancer Cells by a Phenylbutenoid Dimer Isolated from Zingiber cassumunar

Biological and Pharmaceutical Bulletin ◽

10.1248/bpb.30.1561 ◽

2007 ◽

Vol 30 (8) ◽

pp. 1561-1564 ◽

Cited By ~ 15

Author(s):

Jong-Won Lee ◽

Hye-Young Min ◽

Ah-Reum Han ◽

Hwa-Jin Chung ◽

Eun-Jung Park ◽

...

Keyword(s):

Lung Cancer ◽

Cell Cycle ◽

Cancer Cells ◽

Human Lung ◽

Human Lung Cancer ◽

Phase Cell ◽

G1 Phase ◽

Cycle Arrest ◽

Human Lung Cancer Cells ◽

Zingiber Cassumunar

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Viriditoxin Induces G2/M Cell Cycle Arrest and Apoptosis in A549 Human Lung Cancer Cells

Natural Product Sciences ◽

10.20307/nps.2015.21.4.282 ◽

2015 ◽

Vol 21 (4) ◽

pp. 282 ◽

Cited By ~ 15

Author(s):

Ju Hee Park ◽

Tae Hwan Noh ◽

Haibo Wang ◽

Nam Deuk Kim ◽

Jee H. Jung

Keyword(s):

Lung Cancer ◽

Cell Cycle ◽

Cell Cycle Arrest ◽

Cancer Cells ◽

Human Lung ◽

Lung Cancer Cells ◽

Human Lung Cancer ◽

M Cell ◽

Cycle Arrest ◽

Human Lung Cancer Cells

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G2/M cell cycle arrest and induction of apoptosis by a stilbenoid, 3,4,5-trimethoxy-4′-bromo-cis-stilbene, in human lung cancer cells

Life Sciences ◽

10.1016/j.lfs.2004.07.002 ◽

2004 ◽

Vol 75 (23) ◽

pp. 2829-2839 ◽

Cited By ~ 31

Author(s):

Eun-Jin Lee ◽

Hye-Young Min ◽

Hyen Joo Park ◽

Hwa-Jin Chung ◽

Sanghee Kim ◽

...

Keyword(s):

Lung Cancer ◽

Cell Cycle ◽

Cell Cycle Arrest ◽

Cancer Cells ◽

Human Lung ◽

Human Lung Cancer ◽

M Cell ◽

Cycle Arrest ◽

Human Lung Cancer Cells ◽

Induction Of Apoptosis

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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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