IL-1β/NF-κB/ESE3通路在胰腺星型细胞激活及肿瘤耐药中的机制研究

Desmoplastic stroma promotes the development of pancreatic cancer, in which pancreatic stellate cells (PSC) play an important function. With the support of the National Natural Science Foundation of China(81502067), the applicant confirmed that epithelial-specific Ets-3 (ESE3), as a tumor suppressor gene in pancreatic cancer, affects the metastasis of pancreatic cancer (Results published in Cancer Research, PMID: 27923832). However, the expression, function and mechanism of ESE3 in PSC have not been reported yet. Previous work found that ESE3 expression was elevated in PSC of pancreatic cancer tissues, and its expression influenced the prognosis of patients; IL-1β stimulated PSC activation accompanied by NF-κB pathway activation and ESE3 elevated. Therefore, this project will deeply explore the following issues: ① molecular mechanism of ESE3 elevated with PSC activation by IL-1β stimulation；② function of ESE3 on PSC activation；③ the function of IL-1β/ NF-κB/ ESE3 pathway in PSC on Gemcitabine resistance；④ how to increase gemcitabine efficacy by application NF-κB pathway inhibitor. The results not only can enrich the roles of ESE3, but also can provide new strategies for clinical treatment.

间质纤维化促进胰腺癌的发生发展，星型细胞（PSC）在其中发挥重要作用。在国家自然科学基金（81502067）资助下，申请人证实上皮特异性Ets-3（ESE3）在胰腺癌上皮细胞中发挥抑癌基因功能，并影响胰腺癌转移（成果发表在Cancer Research杂志上，PMID：27923832）。但是ESE3在间质PSC中的表达、功能及机制，目前尚无文献报道。前期发现胰腺癌PSC中高表达ESE3，并且其表达水平与患者的预后密切相关；IL-1β活化PSC过程中伴随NF-κB通路激活及ESE3的表达增加。因此本课题拟深入探讨①IL-1β通过NF-κB通路促进PSC中ESE3表达增加的分子机制②ESE3在PSC激活中的具体机制③PSC中IL-1β/NF-κB/ESE3信号通路在吉西他滨耐药中的作用④应用NF-κB通路抑制剂如何增加吉西他滨有效性。这不仅可丰富ESE3的功能，且有望为临床提供新的治疗策略。

方法：通过western blot, RT-PCR, 免疫荧光，流式细胞学检测，染色质免疫共沉淀，双荧光素酶分析，免疫组化和小鼠皮下成瘤实验探讨上皮特异性E26因子3（ESE3）阳性的PSCs在胰腺癌纤维化和化疗耐药中的作用。.结果：与正常的PSCs相比较，ESE3在胰腺癌PSCs中的表达增加。临床数据分析表明PSCs中ESE3的表达与肿瘤大小，pTNM分期，CA19-9,CEA和CA242呈正相关性。PSCs中ESE3与患者的DFS和OS呈负性相关。机制上，PSCs中过表达/降表达ESE3后的上清液影响胰腺癌的化疗效果和肿瘤增殖。ESE3通过与 α-SMA, collagen-I 和 IL-1β启动子区的ESE3结合位点相结合，直接转录调控上述基因的表达。IL-1β通过激活NF-κB通路上调了PSCs中ESE3的表达，并且肿瘤来源的IL-1β通过调节ESE3的表达促进PSCs的激活。.结论：通过抑制IL-1β/ESE3 (PSCs)/IL-1β正反馈环路对于减少胰腺癌的纤维化并增加其化疗效果可能是一个有效的治疗策略。