交感神经发育调节子宫内膜癌发生发展的机制

Autonomic nerve infiltration correlates with poor prognosis in certain tumor progression, and β receptor (sympathetic fibers receptor) blockers can significantly reduce the rate of tumor recurrence and mortality in breast cancer patients. We initially confirmed the expression of tyrosine hydroxylase, a sympathetic fibers marker, was significantly increased in endometrial carcinoma compared to that in normal endometrium. In this study, we would like to demonstrate the regulation of sympathetic development in endometrial cancer. Firstly, tissue microarrays, immunohistochemical analysis and bioinformatic analysis were conducted to clarify the correlation between sympathetic innervation and tumorous grading and staging, the recurrence or survival of patients. Secondly, co-culture of mouse sympathetic ganglion neurons with endometrial cancer cells, follow by MTT assay and transwell assay, was applied to investigate the role of perineural invasion in endometrial cancer cell proliferation, invasion and migration. Then orthotopic endometrial cancer model in nude mice, combined with sympathectomy and bioluminescence technique, was established to study the influence of sympathetic innervation on tumor formation, growth, invasion and metastasis. Furthermore, knockdown of β receptor by RNA interference was used to explore the regulatory mechanisms of sympathetic development mediated by β receptor on endometrial cancer progression. This study is expected to explore a new target for the treatment of endometrial cancer patients, and provide a theoretical basis for clinical diagnosis and treatment of endometrial carcinoma, including nerve-sparing surgery.

自主神经浸润与多种肿瘤的预后密切相关，且β受体（交感神经纤维受体）阻滞剂可显著降低乳腺癌患者的肿瘤复发与致死率。在初步证实内膜癌组织中交感神经纤维标记物酪氨酸羟化酶高表达的基础上，拟阐明交感神经调节在子宫内膜癌发生发展中的作用及机制。首先利用组织芯片、免疫组化与生物信息学统计明确交感神经分布与内膜癌分期（级）、复发率与生存率的相关性；体外共培养小鼠交感神经节神经元与子宫内膜癌细胞，结合MTT、跨膜迁移实验分析交感神经对癌细胞增殖与迁移的影响；进一步建立人子宫内膜癌的裸鼠宫腔原位移植模型，利用交感神经切断术与生物发光技术，体内分析交感神经分布对肿瘤生长、浸润与转移的影响。最后通过RNAi下调β受体表达，深入研究β受体介导交感神经调节内膜癌的作用机制。本研究有望为子宫内膜癌治疗找到新靶点，并为临床诊治（包括保留神经的手术）提供新理论依据。

肿瘤微环境(包括神经浸润)可调控肿瘤生长、浸润与转移，其中神经浸润与多种肿瘤预后不良密切相关。我们初步证实子宫内膜癌组织中神经纤维标记物酪氨酸羟化酶（TH）与神经丝蛋白NEFL的表达显著升高，因而有必要阐明神经调节在子宫内膜癌发生发展中的作用。本研究拟明确神经分布与子宫内膜癌的相关性，阐明神经发育调节子宫内膜癌转移（神经浸润）的作用机制，为指导子宫内膜癌的手术治疗策略提供新的标志物。采用PCR Array分析筛选出子宫内膜癌组织与癌旁组织差异表达的关键神经递质受体（谷氨酸受体2亚型，GRIA2），免疫组织化学/免疫荧光结果显示，子宫内膜癌与不典型增生组织中GRIA2表达高于正常组子宫内膜组织，与新生神经丝蛋白（NEFL）的表达一致，提示子宫内膜癌进展过程中神经在癌变部位的浸润早于肿瘤的转移。原代分离的DRG神经元与Ishikawa细胞共培养可显著促进后者的迁移与侵袭能力，GRIA2抑制剂与shRNA皆可显著抑制这种促转移作用。同时，DRG神经元与Ishikwa共培养可显著促进其培养上清中谷氨酸的浓度。综上，GRIA2介导了DRG神经元的促转移作用，提示其可能作为子宫内膜癌发生神经浸润的潜在标志物，指导临床对II期子宫内膜癌患者行保留神经的广泛性子宫切除术。