Wang H, Tao D, Ma J, Zhang D, Shen Z, Deng C, Zhou J
School of Stomatology, Wannan Medical College, Wuhu 241002, China.
Department of Oral and Maxillofacial Surgery, First Affiliated Hospital (Yijishan Hospital) of Wannan Medical College, Wuhu 241100, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2024 Oct 20;44(10):1947-1954. doi: 10.12122/j.issn.1673-4254.2024.10.13.
To investigate whether cisplatin induces tumor necrosis factor-α (TNF-α) secretion in human head and neck squamous cell carcinoma (HNSCC) cells to trigger RIP1/RIP3/MLKL-dependent necroptosis of the cells.
HNSCC cell lines HN4 and SCC4 treated with cisplatin (CDDP) or the combined treatment with CDDP and z-VAD-fmk (a caspase inhibitor) or Nec-1 (a necroptosis inhibitor) for 24 h were examined for changes in cell viability using CCK8 assay and expressions of caspase-8 and necroptosis pathway proteins (RIP1/RIP3/MLKL) using Western blotting. The changes in migration of the cells were assessed with cell scratch assay, and the expressions of epithelial-mesenchymal transition (EMT) marker proteins N-cadherin, vimentin, and E-cadherin as well as the expressions of NF-κB (p65) and TNF-α were detected with Western blotting.
The IC of cisplatin was 10 μg/mL in HN4 cells and 15 μg/mL in SCC4 cells. Cisplatin treatment significantly decreased the expressions of caspase-8, N-cadherin and vimentin and increased the expressions of Ecadherin, the necroptosis pathway proteins (RIP1/RIP3/MLKL), TNF-α, and NF-κB (p65), and these changes were obviously inhibited by treatment with Nec-1. Cisplatin stimulation also significantly lowered migration of the cells, and this inhibitory effect was strongly attenuated by Nec-1 treatment.
Cisplatin activates nuclear factor-κB signaling in HNSCCs to promote TNF-α autocrine and induce RIP1/RIP3/MLKL-dependent necroptosis, thus leading to inhibition of cell proliferation.
探讨顺铂是否诱导人头颈部鳞状细胞癌（HNSCC）细胞分泌肿瘤坏死因子-α（TNF-α），从而触发细胞的RIP1/RIP3/MLKL依赖性坏死性凋亡。
用顺铂（CDDP）或CDDP与z-VAD-fmk（一种半胱天冬酶抑制剂）或Nec-1（一种坏死性凋亡抑制剂）联合处理HNSCC细胞系HN4和SCC4 24小时，采用CCK8法检测细胞活力变化，采用蛋白质免疫印迹法检测半胱天冬酶-8及坏死性凋亡途径蛋白（RIP1/RIP3/MLKL）的表达。采用细胞划痕试验评估细胞迁移变化，采用蛋白质免疫印迹法检测上皮-间质转化（EMT）标志物蛋白N-钙黏蛋白、波形蛋白和E-钙黏蛋白的表达以及核因子-κB（p65）和TNF-α的表达。
顺铂在HN4细胞中的半数抑制浓度（IC）为10μg/mL，在SCC4细胞中为15μg/mL。顺铂处理显著降低了半胱天冬酶-8、N-钙黏蛋白和波形蛋白的表达，增加了E-钙黏蛋白、坏死性凋亡途径蛋白（RIP1/RIP3/MLKL）、TNF-α和核因子-κB（p65）的表达，而Nec-1处理可明显抑制这些变化。顺铂刺激也显著降低了细胞的迁移能力，而Nec-1处理可强烈减弱这种抑制作用。
顺铂激活HNSCC中的核因子-κB信号通路，促进TNF-α自分泌并诱导RIP1/RIP3/MLKL依赖性坏死性凋亡，从而导致细胞增殖受到抑制。
