Ceramide induces the apoptosis of non‑small cell lung cancer cells through the Txnip/Trx1 complex

Shi,Yining; Jin,Yongmei; Liu,Fangfang; Jiang,Jianjun; Cao,Jiyu; Lu,Youjin; Yang,Jin

doi:10.3892/ijmm.2021.4918

Ceramide induces the apoptosis of non‑small cell lung cancer cells through the Txnip/Trx1 complex

Authors:
- Yining Shi
- Yongmei Jin
- Fangfang Liu
- Jianjun Jiang
- Jiyu Cao
- Youjin Lu
- Jin Yang
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Affiliations: Department of Respiratory Medicine, The Second Hospital of Anhui Medical University, Hefei, Anhui 230022, P.R. China, Department of Respiratory Medicine, The Second People's Hospital of Hefei, Hefei, Anhui 230022, P.R. China, Department of Respiratory Medicine, The First Hospital of Anhui Medical University, Hefei, Anhui 230022, P.R. China, The Teaching Center for Preventive Medicine, School of Public Health, Anhui Medical University, Hefei, Anhui 230022, P.R. China
Published online on: March 19, 2021 https://doi.org/10.3892/ijmm.2021.4918
Article Number: 85
Copyright: © Shi et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Ceramide is a biologically active sphingomyelin that inhibits cell growth and proliferation. In previous studies, it was demonstrated that the use of lipopolysaccharides induces acid sphingomyelinases to produce ceramide, promoting lung cancer cell apoptosis; however, the specific mechanisms of this action remain unclear. Thioredoxin‑interacting protein (Txnip) plays an important role in the signal transmission of redox reactions inside and outside the cell. Thus, it was hypothesized that ceramide induces apoptosis in lung adenocarcinoma cells (A549 and PC9) by modulating the Txnip/Trx1 complex. In the present study, the Cell Counting kit‑8 method was used to detect cell activity and the drug concentration. Hoechst 33258 staining and flow cytometry were used to detect cell apoptosis, and the positional association between Txnip and Trx1 upregulated by ceramide was observed by immunofluorescence confocal microscopy. Reverse transcription‑quantitative polymerase chain reaction and western blot analysis were used to detect the changes in related gene, mRNA and protein expression levels. The results revealed that ceramide treatment resulted in the upregulation of Txnip and in the reduction of Trx1 activities. However, the Txnip inhibitor, verapamil, reversed these changes. The analysis of mRNA expression further verified the changes observed in the protein expression of Txnip, Trx1 and apoptosis‑related proteins. On the whole, the present study demonstrates that ceramide induces the apoptosis of lung cancer cells by regulating the Txnip/Trx1 complex.

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