Department of Pathology, Medical School of Southeast University, Nanjing 210009, China.
Department of Pathology, Medical School of Southeast University, Nanjing 210009, China; Center of Translational Medicine, Zibo Central Hospital, Zibo 255036, China.
Life Sci. 2018 Nov 1;212:182-193. doi: 10.1016/j.lfs.2018.09.033. Epub 2018 Sep 20.
Approximately 20% of head and neck squamous cell carcinomas (HNSCCs) are caused by human papillomavirus (HPV) infection. The effect of arsenic trioxide (ATO) on HPV oncogene expression of HNSCC cells remains unknown. In this study, we investigated the anti-cancer activity and possible molecular pathways of ATO on the six HNSCC cell lines (three HPV-positive and three HPV-negative).
The effects of ATO on the cell proliferation, apoptosis, cell cycle of HNSCC cells were analyzed using CCK-8 assay, colony formation and flow cytometry. Transwell assay was used to examine the effect of ATO on cell migration. The transcriptional and protein expression of key genes were determined by real-time PCR and Western blot, respectively. Using a xenograft model, we assessed the effects of ATO on HNSCC cells in vivo.
HPV-positive and -negative HNSCC cells had different expression of key genes. ATO inhibited HNSCC cell proliferation and migration and induced apoptosis and these effects were more significant in HPV-positive HNSCC cells than in HPV-negative HNSCC cells. ATO treatment reduced the expression of HPV16-E6/E7 and cyclin D1 proteins and enhanced the expression of p16, pRb, and p53 in HPV-positive HNSCC cells. By contrast, ATO treatment reduced the expression of epidermal growth factor receptor, cyclin D1 and mutant p53 and enhanced the expression of pRb in HPV-negative HNSCC cells. Anti-cancer effect of ATO on HNSCCs was confirmed by inhibiting xenograft growth in vivo.
Our data suggest that ATO is a potential therapeutic drug for HNSCCs, especially HPV-positive HNSCCs.
大约 20%的头颈部鳞状细胞癌（HNSCC）是由人乳头瘤病毒（HPV）感染引起的。三氧化二砷（ATO）对 HNSCC 细胞 HPV 癌基因表达的影响尚不清楚。在这项研究中，我们研究了 ATO 对六种 HNSCC 细胞系（三种 HPV 阳性和三种 HPV 阴性）的抗癌活性和可能的分子途径。
通过 CCK-8 测定、集落形成和流式细胞术分析 ATO 对 HNSCC 细胞增殖、凋亡、细胞周期的影响。Transwell 测定用于检测 ATO 对细胞迁移的影响。通过实时 PCR 和 Western blot 分别测定关键基因的转录和蛋白表达。通过异种移植模型，我们评估了 ATO 对体内 HNSCC 细胞的影响。
HPV 阳性和阴性 HNSCC 细胞关键基因的表达不同。ATO 抑制 HNSCC 细胞增殖和迁移，并诱导凋亡，这些效应在 HPV 阳性 HNSCC 细胞中比在 HPV 阴性 HNSCC 细胞中更为显著。ATO 处理降低了 HPV16-E6/E7 和细胞周期蛋白 D1 蛋白的表达，并增强了 HPV 阳性 HNSCC 细胞中 p16、pRb 和 p53 的表达。相比之下，ATO 处理降低了 HPV 阴性 HNSCC 细胞中表皮生长因子受体、细胞周期蛋白 D1 和突变型 p53 的表达，并增强了 pRb 的表达。体内抑制异种移植生长证实了 ATO 对 HNSCC 的抗癌作用。
我们的数据表明，ATO 是 HNSCC 的一种潜在治疗药物，特别是 HPV 阳性 HNSCC。
