Limonene terpenoid obstructs human bladder cancer cell (T24 cell line) growth by inducing cellular apoptosis, caspase activation, G2/M phase cell cycle arrest and stops cancer metastasis. 2020

Zhaohua Ye, and Zhulin Liang, and Qiwu Mi, and Yonglian Guo
Department of Urology, Affiliated Dongguan People's Hospital, Southern Medical University, Dongguan 523059, Guangdong, P.R. China.

OBJECTIVE The primary purpose of the current study was to investigate the antitumor activity of limonene which is a plant monoterpene along with evaluating its effects on cell apoptosis, cell cycle phase distribution, cell migration and invasion. METHODS The cell proliferation of T24 bladder cancer cells was examined by WTS-1 assay. The apoptotic effects induced by limonene were investigated by a combination of fluorescence microscopy and flow cytometry and then confirmed by western blot assay. The effects of limonene on cell cycle in T24 bladder cancer cells were studied by flow cytometry. The effects on cell migration and invasion were examined by wound healing assay and transwell assay using Matrigel. RESULTS The results showed that limonene induced cytotoxic effects and reduced cell viability of T24 human bladder cancer cells showing an IC50 value of 9 μM. Limonene also induced significant apoptosis in bladder cancer cells since it induced significant nuclear fragmentation, chromatin condensation, and splitting of the nucleus, representative of the apoptotic cascade. The apoptotic cell percentage was 1.95, 5.35, 15.61 and 34.71% at limonene concentrations of 0, 9, 18 and 36 µM. Further, the apoptotic effects of limonene were also confirmed by Western blot analysis and the results showed increase in the expression of Bax and caspase-3 and decrease of Bcl-2 expression. Limonene also caused G2/M phase cell cycle arrest as well as suppression of cell migration and invasion. CONCLUSIONS These results indicate that limonene might be used as a potent anticancer agent against human bladder cancer for which further in depth studies are needed, especially over its toxicological studies.

UI MeSH Term Description Entries
D009362 Neoplasm Metastasis The transfer of a neoplasm from one organ or part of the body to another remote from the primary site. Metastase,Metastasis,Metastases, Neoplasm,Metastasis, Neoplasm,Neoplasm Metastases,Metastases
D001749 Urinary Bladder Neoplasms Tumors or cancer of the URINARY BLADDER. Bladder Cancer,Bladder Neoplasms,Cancer of Bladder,Bladder Tumors,Cancer of the Bladder,Malignant Tumor of Urinary Bladder,Neoplasms, Bladder,Urinary Bladder Cancer,Bladder Cancers,Bladder Neoplasm,Bladder Tumor,Cancer, Bladder,Cancer, Urinary Bladder,Neoplasm, Bladder,Neoplasm, Urinary Bladder,Tumor, Bladder,Tumors, Bladder,Urinary Bladder Neoplasm
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D000077222 Limonene A naturally-occurring class of MONOTERPENES which occur as a clear colorless liquid at room temperature. Limonene is the major component in the oil of oranges which has many uses, including as flavor and fragrance. It is recognized as safe in food by the Food and Drug Administration (FDA). (+)-(R)-4-isopropenyl-1-methylcyclohexene,(+)-Limonene,(-)-Limonene,(4R)-1-methyl-4-(1-methylethenyl)cyclohexene,(4S)-1-methyl-4-isopropenylcyclohex-1-ene,(D)-Limonene,(R)-(+)-Limonene,(R)-4-isopropenyl-1-methylcyclohexene,1-Methyl-4-(1-methylethenyl)cyclohexene,4-Mentha-1,8-diene,AISA 5203-L (+)Limonene,Cyclohexene, 1-methyl-4-(1-methylethenyl)-, (4R)-,Dipentene,Limonene, (+)-,Limonene, (+-)-,Limonene, (+-)-isomer,Limonene, (R)-isomer,Limonene, (S)-isomer,d-Limonene,4 Mentha 1,8 diene,d Limonene
D013729 Terpenes A class of compounds composed of repeating 5-carbon units of HEMITERPENES. Isoprenoid,Terpene,Terpenoid,Isoprenoids,Terpenoids
D015398 Signal Transduction The intracellular transfer of information (biological activation/inhibition) through a signal pathway. In each signal transduction system, an activation/inhibition signal from a biologically active molecule (hormone, neurotransmitter) is mediated via the coupling of a receptor/enzyme to a second messenger system or to an ion channel. Signal transduction plays an important role in activating cellular functions, cell differentiation, and cell proliferation. Examples of signal transduction systems are the GAMMA-AMINOBUTYRIC ACID-postsynaptic receptor-calcium ion channel system, the receptor-mediated T-cell activation pathway, and the receptor-mediated activation of phospholipases. Those coupled to membrane depolarization or intracellular release of calcium include the receptor-mediated activation of cytotoxic functions in granulocytes and the synaptic potentiation of protein kinase activation. Some signal transduction pathways may be part of larger signal transduction pathways; for example, protein kinase activation is part of the platelet activation signal pathway. Cell Signaling,Receptor-Mediated Signal Transduction,Signal Pathways,Receptor Mediated Signal Transduction,Signal Transduction Pathways,Signal Transduction Systems,Pathway, Signal,Pathway, Signal Transduction,Pathways, Signal,Pathways, Signal Transduction,Receptor-Mediated Signal Transductions,Signal Pathway,Signal Transduction Pathway,Signal Transduction System,Signal Transduction, Receptor-Mediated,Signal Transductions,Signal Transductions, Receptor-Mediated,System, Signal Transduction,Systems, Signal Transduction,Transduction, Signal,Transductions, Signal
D017209 Apoptosis A regulated cell death mechanism characterized by distinctive morphologic changes in the nucleus and cytoplasm, including the endonucleolytic cleavage of genomic DNA, at regularly spaced, internucleosomal sites, i.e., DNA FRAGMENTATION. It is genetically programmed and serves as a balance to mitosis in regulating the size of animal tissues and in mediating pathologic processes associated with tumor growth. Apoptosis, Extrinsic Pathway,Apoptosis, Intrinsic Pathway,Caspase-Dependent Apoptosis,Classic Apoptosis,Classical Apoptosis,Programmed Cell Death,Programmed Cell Death, Type I,Apoptoses, Extrinsic Pathway,Apoptoses, Intrinsic Pathway,Apoptosis, Caspase-Dependent,Apoptosis, Classic,Apoptosis, Classical,Caspase Dependent Apoptosis,Cell Death, Programmed,Classic Apoptoses,Extrinsic Pathway Apoptoses,Extrinsic Pathway Apoptosis,Intrinsic Pathway Apoptoses,Intrinsic Pathway Apoptosis
D045744 Cell Line, Tumor A cell line derived from cultured tumor cells. Tumor Cell Line,Cell Lines, Tumor,Line, Tumor Cell,Lines, Tumor Cell,Tumor Cell Lines
D059565 G2 Phase Cell Cycle Checkpoints CELL CYCLE regulatory signaling systems that are triggered by DNA DAMAGE or lack of nutrients during G2 PHASE. When triggered they restrain cells transitioning from G2 phase to M PHASE. Antephase Checkpoint,G2 Cell Cycle Arrest,G2 Cell Cycle Checkpoints,G2 Cell Cycle Control,G2 Phase Checkpoints,G2 Restriction Point,G2-M Cell Cycle Checkpoint,Antephase Checkpoints,Checkpoint, Antephase,Checkpoint, G2 Phase,Checkpoints, Antephase,Checkpoints, G2 Phase,G2 M Cell Cycle Checkpoint,G2 Phase Checkpoint,G2 Restriction Points,Restriction Point, G2,Restriction Points, G2
D059566 M Phase Cell Cycle Checkpoints The cellular signaling system that halts the progression of cells through MITOSIS or MEIOSIS if a defect that will affect CHROMOSOME SEGREGATION is detected. M Phase Cell Cycle Checkpoint,M Phase Cell Cycle Arrest,M Phase Checkpoint,Mitotic Checkpoints,Mitotic Spindle Checkpoints,Spindle Assembly Checkpoint,Spindle Assembly Checkpoint Network,Assembly Checkpoint, Spindle,Assembly Checkpoints, Spindle,Checkpoint, M Phase,Checkpoint, Mitotic,Checkpoint, Mitotic Spindle,Checkpoint, Spindle Assembly,Checkpoints, M Phase,Checkpoints, Mitotic,Checkpoints, Mitotic Spindle,Checkpoints, Spindle Assembly,M Phase Checkpoints,Mitotic Checkpoint,Mitotic Spindle Checkpoint,Phase Checkpoint, M,Spindle Assembly Checkpoints,Spindle Checkpoint, Mitotic,Spindle Checkpoints, Mitotic

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