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A synthetic coumarin derivative (4‐flourophenylacetamide‐acetyl coumarin) impedes cell cycle at G0/G1 stage, induces apoptosis, and inhibits metastasis via ROS‐mediated p53 and AKT signaling pathways in A549 cells
Author(s) -
Umar Shweta,
Soni Rina,
Durgapal Sunil D.,
Soman Subhangi,
Balakrishnan Suresh
Publication year - 2020
Publication title -
journal of biochemical and molecular toxicology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.526
H-Index - 58
eISSN - 1099-0461
pISSN - 1095-6670
DOI - 10.1002/jbt.22553
Subject(s) - cytotoxicity , a549 cell , chemistry , apoptosis , acridine orange , cell cycle , cancer research , pi3k/akt/mtor pathway , protein kinase b , cell cycle checkpoint , microbiology and biotechnology , biology , biochemistry , in vitro
Abstract New chemotherapeutic agents with minimum side effects are indispensable to treat non–small‐cell lung cancer (NSCLC) since the mortality rate of patients suffering from NSCLC remains high despite receiving conventional medication. In our previous study, many coumarin derivatives were screened for their anticancer properties in A549, an in vitro NSCLC model. One of these, 4‐flourophenylacetamide‐acetyl coumarin (4‐FPAC), induced cytotoxicity at a concentration as low as 0.16 nM. Herein, initially, the cytotoxic potential of 4‐FPAC was tested on a noncancerous cell line NIH3T3 and was found safe at the selected dose of 0.16 nM. Further, we investigated the mechanism by which 4‐FPAC induced cytotoxicity and arrested the progression of cell cycle as well as metastasis in A549. Results of ethidium bromide/acridine orange (EtBr/AO), 4,6‐diamidino‐2‐phenylindole, comet, and lactate dehydrogenase assays revealed that 4‐FPAC caused cytotoxicity via reactive oxygen species‐induced p53‐mediated mechanism, which involves both extrinsic and intrinsic pathways of apoptosis. Dichlorodihydrofluorescein diacetate, rhodamine 123, and AO staining confirmed the involvement of both mitochondria and lysosome in inducing apoptosis. However, flow cytometric analysis revealed that it causes cell cycle arrest at the G0/G1 phase by modulating p21, CDK2, and CDK4 expression. Aggregation, soft‐agar, clonogenic, and scratch assays as well as gene expression analysis collectively confirmed that 4‐FPAC minimizes the metastatic property of A549 by downregulating Snail, matrix metalloproteinase 9, and interleukin‐8. Additional studies reaffirmed the above findings and substantiated the role of PI3K/AKT in achieving them. The cell‐type‐specific selective cytostatic and antimetastatic properties shown by 4‐FPAC indicate its potential to emerge as a drug of choice against NSCLC in the future.