CENPN suppresses autophagy and increases paclitaxel resistance in nasopharyngeal carcinoma cells by inhibiting the CREB-VAMP8 signaling axis.
CENPN suppresses autophagy and increases paclitaxel resistance in nasopharyngeal carcinoma cells by inhibiting the CREB-VAMP8 signaling axis.
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DOI：
10.1080/15548627.2023.2258052
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发表时间：
2024-02
期刊：
影响因子：
13.3
通讯作者：
Chen, Shi-Ming
中科院分区：
文献类型：
--
作者：
Wang, Bin-Ru;Han, Ji-Bo;Jiang, Yang;Xu, Shan;Yang, Rui;Kong, Yong-Gang;Tao, Ze-Zhang;Hua, Qing-Quan;Zou, You;Chen, Shi-Ming
化疗耐药是鼻咽癌患者预后不良的主要原因之一。我们发现CENPN能促进鼻咽癌细胞的生长、增殖和凋亡抵抗，但其与鼻咽癌化疗耐药的关系尚不清楚。本研究通过对临床病例的分析，证实鼻咽癌患者CENPN表达水平与紫杉醇（PTX）耐药程度及预后不良呈正相关。在通过转录组测序敲低CENPN后，VAMP 8表达显著增加。我们在细胞实验中发现，CENPN抑制巨自噬/自噬和VAMP 8表达，并显着增加PTX抗性。过表达CENPN可通过抑制自噬来降低PTX对鼻咽癌细胞存活、增殖、细胞周期进程和凋亡抵抗的抑制作用。反过来，CENPN的敲低可以通过增加自噬来影响NPC细胞的表型以实现PTX敏化。连续敲除CENPN和VAMP 8逆转了单独敲除CENPN的PTX致敏作用。裸鼠实验证实，敲低CENPN可增加VAMP 8表达，增强自噬，增加NPC细胞对PTX的敏感性。机制研究表明，CENPN抑制p-CREB易位到NPC细胞的细胞核中，导致p-CREB与VAMP 8启动子的结合减少，从而抑制VAMP 8的转录。这些结果表明，CENPN可能是一个标志物，用于预测化疗疗效和潜在的目标，诱导化疗敏感剂，如PTX。缩略语：3-甲基丙烯酸：3-甲基腺嘌呤; ATG5：自噬相关5例; CENPN：着丝粒蛋白N; CQ：氯喹; CREB：cAMP反应元件结合蛋白; ChIP：染色质免疫沉淀试验; IC 50：半数最大抑制浓度; LAMP 2A：溶酶体相关膜蛋白2A; MAP 1 LC 3/LC 3：微管相关蛋白1轻链3; NPC：鼻咽癌; NPG：鼻咽炎; oeCENPN：过表达CENPN; PTX：紫杉醇; RAPA：雷帕霉素; RNA-seq：转录组测序; shCENPN：靶向人CENPN基因的小发夹RNA表达载体; shCENPN-shVAMP 8：靶向人CENPN基因和VAMP 8基因的顺序敲低; shVAMP 8：靶向人VAMP 8基因的小发夹RNA表达载体; TEM：透射电子显微镜; TIR：肿瘤抑制率; VAMP 8：囊泡相关膜蛋白8。
Chemotherapeutic resistance is one of the most common reasons for poor prognosis of patients with nasopharyngeal carcinoma (NPC). We found that CENPN can promote the growth, proliferation and apoptosis resistance of NPC cells, but its relationship with chemotherapeutic resistance in NPC is unclear. Here we verified that the CENPN expression level in NPC patients was positively correlated with the degree of paclitaxel (PTX) resistance and a poor prognosis through analysis of clinical cases. VAMP8 expression was significantly increased after knockdown of CENPN by transcriptome sequencing. We found in cell experiments that CENPN inhibited macroautophagy/autophagy and VAMP8 expression and significantly increased PTX resistance. Overexpression of CENPN reduced the inhibitory effects of PTX on survival, cell proliferation, cell cycle progression and apoptosis resistance in NPC cells by inhibiting autophagy. In turn, knockdown of CENPN can affect the phenotype of NPC cells by increasing autophagy to achieve PTX sensitization. Sequential knockdown of CENPN and VAMP8 reversed the PTX-sensitizing effect of CENPN knockdown alone. Experiments in nude mice confirmed that knockdown of CENPN can increase VAMP8 expression, enhance autophagy and increase the sensitivity of NPC cells to PTX. Mechanistic studies showed that CENPN inhibited the translocation of p-CREB into the nucleus of NPC cells, resulting in the decreased binding of p-CREB to the VAMP8 promoter, thereby inhibiting the transcription of VAMP8. These results demonstrate that CENPN may be a marker for predicting chemotherapeutic efficacy and a potential target for inducing chemosensitization to agents such as PTX. Abbreviations: 3-MA: 3-methyladenine; ATG5: autophagy related 5; CENPN: centromere protein N; CQ: chloroquine; CREB: cAMP responsive element binding protein; ChIP: chromatin immunoprecipitation assay; IC50: half-maximal inhibitory concentration; LAMP2A: lysosomal associated membrane protein 2A; MAP1LC3/LC3: microtubule associated protein 1 light chain 3; NPC: nasopharyngeal carcinoma; NPG: nasopharyngitis; oeCENPN: overexpressed CENPN; PTX: paclitaxel; RAPA: rapamycin; RNA-seq: transcriptome sequencing; shCENPN: small hairpin RNA expression vector targeting the human CENPN gene; shCENPN-shVAMP8: sequential knockdown targeting the human CENPN gene and VAMP8 gene; shVAMP8: small hairpin RNA expression vector targeting the human VAMP8 gene; TEM: transmission electron microscopy; TIR: tumor inhibitory rate; VAMP8: vesicle associated membrane protein 8.
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影响因子：
16.6
作者：
Huang H;Ouyang Q;Zhu M;Yu H;Mei K;Liu R
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DOI：
10.1186/s13046-021-02146-8
发表时间：
2021-10-27
期刊：
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