目的：探讨扁柏酚对鼻咽癌细胞CNE1细胞周期和凋亡的影响及相关分子机制。方法：体外培养CNE1细胞，与不同浓度和厚朴酚孵育，将细胞分为空白对照组、10 μmol/L、20 μmol/L、40 μmol/L和厚朴酚处理组以及10 μg/ml和厚朴酚处理组。顺铂组。采用甲基噻唑基二苯基溴化四唑（MTT）法测定细胞活力，流式细胞仪检测细胞周期分布，线粒体膜电位检测试剂盒检测线粒体膜电位，末端脱氧核苷酸转移酶介导的dUTP缺口末端标记（TUNEL）检测细胞凋亡）方法，采用免疫印迹法检测增殖细胞核抗原（PCNA）和G1/S特异性细胞周期蛋白D1（cyclin D1）的蛋白表达。 RNA-Seq 在经扁柏酚处理的细胞中进行。通过定量逆转录聚合酶链反应（RT-qPCR）检测yes相关蛋白δ（YAP）的mRNA表达。免疫印迹法检测磷酸YAP（p-YAP）和核YAP蛋白的表达，免疫荧光法检测经或未经哺乳动物STE20样激酶1/2（MST1/ 2)抑制剂(XMU-MP-1)、扁柏酚和XMU-MP-1扁柏酚。使用GraphPad Prism 8.0软件对数据进行统计分析。结果 与对照组相比，扁柏酚处理组CNE1细胞的细胞活力、线粒体膜电位水平、PCNA和cyclin D1蛋白表达均明显降低（均P值<0.05），而G0/ G1期细胞和TUNEL阳性细胞比例均显着增加（P值均<0.05）。转录组分析显示差异基因主要富集于Wnt信号通路、肿瘤坏死因子通路、Hippo信号通路。扁柏酚处理细胞中YAP mRNA水平和细胞核YAP蛋白表达水平降低，p-YAP蛋白水平升高，与对照组比较差异有统计学意义（均P值<0.05）。与扁柏酚组相比，XMU-MP-1扁柏酚组p-YAP蛋白表达量减少（1.157±0.076 vs 0.479±0.038，t=37.120，P<0.05），细胞核内YAP蛋白表达量增加（ 0.143±0.012 vs 0.425±0.031，t=29.181，P<0.05)，G0/G1期细胞比例减少[(72.494±3.309)% vs (58.747±2.865)%，t=17.265，P<0.05] ，凋亡率降低[(53.158±3.376)% vs (29.621±2.713)%，t=28.584，P<0.05]。结论：扁柏酚可通过Hippo/YAP信号通路阻滞CNE1细胞的细胞周期并诱导细胞凋亡。

Objective: To explore the effects of hinokiol on the cell cyle and apoptosis of CNE1 nasopharyngeal carcinoma cells and the relevant molecular mechanism. Methods: The CNE1 cells were cultured in vitro and incubated with different concentrations of honokiol, and the cells were divided into blank control group, 10 μmol/L, 20 μmol/L and 40 μmol/L hinokiol treatment groups, and 10 μg/ml cisplatin group. Cell viability was determined by methylthiazolyldiphenyl- tetrazolium bromide (MTT) method, the cell cycle distribution was detected by flow cytometry, mitochondrial membrane potential was detected by mitochondrial membrane potential test kit, apoptosis was detected by terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) method, and the proteins expression of proliferating cell nuclear antigen (PCNA) and G1/S specific cyclin D1 (cyclin D1) were detected by immunoblotting. RNA-Seq was conducted in the hinokiol-treated cells. The mRNA expression of yes-associated protein delta (YAP) was detected by quantitative reverse transcription polymerase chain reaction (RT-qPCR). The proteins expression of phosphor-YAP (p-YAP) and nuclear YAP were detected by immunoblotting, the nuclear distribution of YAP protein was detected by immunofluorescence in the cells with or without treated with the mammalian STE20-like kinase 1/2 (MST1/2) inhibitor (XMU-MP-1), hinokiol, and XMU-MP-1+hinokiol. Statistical analysis of the data was conducted using GraphPad Prism 8.0 software. Resluts Compared with the control group, the cell viablity of CNE1 cells, the levels of mitochondrial membrane potential, the proteins expression of PCNA and cyclin D1 in hinokiol treatment groups were markedly decreased (all P values<0.05), while the proportion of G0/G1 phase cells and the ratio of TUNEL-positive cells were significantly increased (both P values<0.05). Transcriptome analysis showed that differential genes were mainly enriched in Wnt signaling pathway, tumor necrosis factor pathway, and Hippo signaling pathway. The mRNA level of YAP and the protein expression of YAP in the nucleus were decreased and the level of p-YAP protein was increased in cells treated with hinokiol, which were significantly different from control group (all P values<0.05). Compared with the hinokiol group, XMU-MP-1+hinokiol groups showed the decrease of p-YAP protein expression (1.157±0.076 vs 0.479±0.038, t=37.120, P<0.05), the increase of YAP protein expression in the nucleus (0.143±0.012 vs 0.425±0.031, t=29.181, P<0.05), the reduced proportion of cells in G0/G1 phase [(72.494±3.309)% vs (58.747±2.865)%, t=17.265, P<0.05], and the decrease of apoptosis ratio [(53.158±3.376)% vs (29.621±2.713)%, t=28.584, P<0.05]. Conclusion: Hinokiol can arrest the cell cycle and induce the cell apoptosis of CNE1 cells via Hippo/YAP signaling pathway.