PRKRA通过NF-κB途径上调MMP1转录促进胰腺癌进展。
PRKRA promotes pancreatic cancer progression by upregulating MMP1 transcription via the NF-κB pathway.
作者信息
Qiu Jiangdong, Feng Mengyu, Yang Gang, Su Dan, Zhao Fangyu, Liu Yueze, Tao Jinxin, Luo Wenhao, Zhang Taiping
机构信息
Department of General Surgery, State Key Laboratory of Complex Severe and Rare Diseases, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, 100730, China.
Key Laboratory of Research in Pancreatic Tumor, Chinese Academy of Medical Sciences, Beijing, 100730, China.
出版信息
Heliyon. 2023 Jun 10;9(6):e17194. doi: 10.1016/j.heliyon.2023.e17194. eCollection 2023 Jun.
OBJECTIVE
Pancreatic cancer (PC) is highly malignant, but the underlying mechanisms of cancer progression remain unclear. PRKRA is involved in cellular stress response, but its role in PC was unknown.
METHODS
The expression of PRKRA between normal and tumor tissues were compared, and the prognostic value of PRKRA was evaluated. SiRNA and plasmids were applied to investigate the effects of PRKRA on PC cells. Organoids and cell lines with knockout and overexpression of PRKRA were established by CRISPR/Cas9 and lentivirus. The effects of PRKRA on PC were evaluated in vivo by cell-derived xenografts. The downstream genes of PRKRA were screened by transcriptome sequencing. The regulation of the target gene was validated by RT-qPCR, western blot, ChIP and dual luciferase reporter assay. Besides, the correlation between PRKRA and gemcitabine sensitivity was investigated by PC organoids.
RESULTS
PRKRA was significantly overexpressed in PC tissues and independently associated with poor prognosis. PRKRA promoted the proliferation, migration, and chemoresistance of PC cells. The proliferation of PC organoids was decreased by PRKRA knockout. The growth and chemoresistance of xenografts were increased by PRKRA overexpression. Mechanistically, PRKRA upregulated the transcription of MMP1 via NF-κB pathway. ChIP and dual luciferase reporter assay showed that NF-κB subunit P65 could bind to the promoter of MMP1. The sensitivity of PC organoids to gemcitabine was negatively correlated with the expression of PRKRA and MMP1.
CONCLUSIONS
Our study indicated that the PRKRA/NF-κB/MMP1 axis promoted the progression of PC and may serve as a potential therapeutic target and prognosis marker.
目的
胰腺癌（PC）具有高度恶性，但癌症进展的潜在机制仍不清楚。PRKRA参与细胞应激反应，但其在PC中的作用尚不清楚。
方法
比较正常组织和肿瘤组织中PRKRA的表达，并评估PRKRA的预后价值。应用小干扰RNA（SiRNA）和质粒研究PRKRA对PC细胞的影响。通过CRISPR/Cas9和慢病毒建立PRKRA敲除和过表达的类器官和细胞系。通过细胞来源的异种移植在体内评估PRKRA对PC的影响。通过转录组测序筛选PRKRA的下游基因。通过逆转录定量聚合酶链反应（RT-qPCR）、蛋白质免疫印迹法（western blot）、染色质免疫沉淀法（ChIP）和双荧光素酶报告基因检测验证靶基因的调控。此外，通过PC类器官研究PRKRA与吉西他滨敏感性之间的相关性。
结果
PRKRA在PC组织中显著过表达，且与不良预后独立相关。PRKRA促进PC细胞的增殖、迁移和化疗耐药性。PRKRA敲除降低了PC类器官的增殖。PRKRA过表达增加了异种移植瘤的生长和化疗耐药性。机制上，PRKRA通过核因子κB（NF-κB）途径上调基质金属蛋白酶1（MMP1）的转录。ChIP和双荧光素酶报告基因检测表明，NF-κB亚基P65可与MMP1的启动子结合。PC类器官对吉西他滨的敏感性与PRKRA和MMP1的表达呈负相关。
结论
我们的研究表明，PRKRA/NF-κB/MMP1轴促进了PC的进展，可能作为潜在的治疗靶点和预后标志物。
Zotero 插件