Isoliquiritigenin induces apoptosis of human bladder cancer T24 cells via a cyclin‑dependent kinase‑independent mechanism Corrigendum in /10.3892/ol.2021.12529

Si,Lingling; Yang,Xinhui; Yan,Xinyan; Wang,Yanming; Zheng,Qiusheng

doi:10.3892/ol.2017.6159

Isoliquiritigenin induces apoptosis of human bladder cancer T24 cells via a cyclin‑dependent kinase‑independent mechanism

Corrigendum in: /10.3892/ol.2021.12529

Authors:
- Lingling Si
- Xinhui Yang
- Xinyan Yan
- Yanming Wang
- Qiusheng Zheng
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Affiliations: School of Pharmacy, Shihezi University, Shihezi, Xinjiang 832002, P.R. China, Department of Pharmacy, The First Affiliated Hospital of the Medical College, Xinjiang Shihezi University, Shihezi, Xinjiang 832002, P.R. China
Published online on: May 12, 2017 https://doi.org/10.3892/ol.2017.6159
Pages: 241-249
Copyright: © Si et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The aim of the present study was to investigate whether an increase in cyclin‑dependent kinase 2 (CDK2) activity is involved in apoptosis of human bladder cancer T24 cells induced by isoliquiritigenin (ISL). The viability of T24 cells was estimated using a sulforhodamine B assay. Cell morphological changes were examined using Hoechst 33258 staining. The apoptotic rate was determined by staining cells with Annexin V‑fluorescein isothiocyanate and propidium iodide labeling. The mitochondrial membrane potential (ΔΨm) was measured using 5,5',6,6'‑tetrachloro‑1,1', 3,3'‑tetraethyl benzimidazole carbocyanine iodide. Alterations in the apoptosis‑related regulators B‑cell lymphoma‑2 (Bcl‑2), Bcl‑2‑associated X protein (Bax), Bcl‑2‑interacting mediator of cell death (Bim), apoptotic protease‑activating facter‑1 (Apaf‑1), caspase‑9 and caspase‑3 were determined using reverse transcription‑polymerase chain reaction (PCR) and quantitative PCR methods. Western blot analysis was used to detect the expression of Bcl‑2, Bax and caspase‑3. CDK2 activity was measured using a spectrometric assay. Following treatment with ISL (between 30 and 70 µg/ml) for 24 h, typical apoptotic morphological changes were observed in T24 cells, exhibiting an edge set of chromosomes, nuclear condensation, nuclear fragmentation and other morphological features. Treatment with ISL increased the apoptotic ratio of T24 cells in a concentration‑dependent manner and induced a decrease in the ΔΨm in a time‑dependent manner. Treatment with ISL upregulated the expression of Bax, Bim, Apaf‑1, caspase‑9 and caspase‑3, downregulated the expression of Bcl‑2, and increased CDK2 activity. MK‑8776 (an inhibitor of CDK2) antagonized the apoptosis induced by ISL, and, compared with treatment with ISL alone, pretreatment with MK‑8776 inhibited the decrease in ΔΨm, downregulated the mRNA expression of Bax, Bim, Apaf‑1, caspase‑9 and caspase‑3, and upregulated Bcl‑2 mRNA expression. Western blot analysis demonstrated that, with increasing ISL concentration, the Bcl‑2 expression level was significantly decreased (P<0.05), whereas caspase‑3 and Bax expression levels were significantly increased (P<0.01). These results indicated that ISL treatment caused a significant decrease in the proliferation rate and increase in apoptosis of T24 cells. The mechanism by which ISL induces T24 cell apoptosis in vitro may be associated with an increase in CDK2 activity, downregulation of the ΔΨm and activation of caspase‑3/caspase‑9‑mediated mitochondrial apoptotic signaling pathways.

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