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.
Int J Mol Med. 2021 May;47(5). doi: 10.3892/ijmm.2021.4918. Epub 2021 Mar 24.
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.
神经酰胺是一种具有生物活性的鞘磷脂,能抑制细胞生长和增殖。在之前的研究中,已经证明使用脂多糖可诱导酸性鞘磷脂酶产生神经酰胺,从而促进肺癌细胞凋亡;然而,其具体作用机制尚不清楚。硫氧还蛋白相互作用蛋白(Txnip)在细胞内外的氧化还原反应信号转导中发挥着重要作用。因此,研究人员假设神经酰胺通过调节 Txnip/Trx1 复合物诱导肺腺癌细胞(A549 和 PC9)凋亡。在本研究中,采用细胞计数试剂盒-8 法检测细胞活性和药物浓度。采用 Hoechst 33258 染色和流式细胞术检测细胞凋亡,并通过免疫荧光共聚焦显微镜观察神经酰胺上调的 Txnip 和 Trx1 的位置关联。采用反转录-定量聚合酶链反应和 Western blot 分析检测相关基因、mRNA 和蛋白表达水平的变化。结果表明,神经酰胺处理导致 Txnip 上调和 Trx1 活性降低。然而,Txnip 抑制剂维拉帕米逆转了这些变化。mRNA 表达分析进一步验证了 Txnip、Trx1 和凋亡相关蛋白的蛋白表达变化。总的来说,本研究表明神经酰胺通过调节 Txnip/Trx1 复合物诱导肺癌细胞凋亡。