miR34a/FNDC3B调控肿瘤干细胞样细胞干性影响食管癌浸润转移的机制研究

Esophageal cancer（EC） is a common malignant tumor of the digestive system, and the incidence of Esophageal cancer of kazaks which located in the northern xinjiang was significantly higher than other ethnic groups, high properties of invasion and metastasis is one of the leading causes of death among patients with EC;A new study has found that cancer stem cells is closely related to infiltration and metastasis of tumor, we will Take the EC population in xinjiang which the high incidence area of EC as research objects, and using RNA sequencing, bioinformatics software, CO-IP method and Western blot technology screening and verification of invasion and metastasis related signaling pathway proteins that regulation by FNDC3B, and then respectively in histology, cytology level in vitro and in vivo zoology levels, using the immunohistochemical technique, plasmid transfection, nude mouse transplantation tumor model building to detect the expression of miR34a/FNDC3B and related pathway molecules which Downstream invasion and metastasis, cancer stem cells markers and Drug resistance molecular, observation the possible regulatory mechanism which the influence of miR34a/FNDC3B on esophageal cell/stem-like cells and involved in the invasion and metastasis of esophageal cancer and chemotherapy resistance, providing the new ideas for the development of genetic level and the accuracy therapy of molecular targeted and the choice of treatment strategy and improve of the prognosis targeted cancer stem cells for xinjiang kazaks esophageal cancer.

食管癌是消化系统常见恶性肿瘤，位于新疆北部的哈萨克族（哈族）食管鳞癌发病率明显高于同地区其他民族，食管癌的高侵袭和转移特性是患者死亡的主要原因之一，最新研究发现肿瘤干细胞与肿瘤的浸润转移密切相关；本项目拟以新疆食管癌高发区人群为研究对象，运用RNA测序、生物信息学软件、CO-IP方法及Western blot技术筛选及验证FNDC3B调控的侵袭转移相关信号通路分子，然后分别在组织学水平、体外细胞学水平和体内动物学水平运用免疫组织化学技术、质粒转染、裸鼠移植瘤模型构建等方法检测miR34a/FNDC3B及FNDC3B调控的侵袭转移相关通路分子和食管癌干性相关分子及耐药相关分子的表达情况，观察miR34a/FNDC3B及其调控的侵袭转移相关通路分子对食管癌细胞/干细胞样细胞干性及侵袭转移等生物学行为的影响及可能调控机制，分析、探讨miR34a/FNDC3B影响食管癌侵袭转移及化疗抵抗的可能机制

食管癌是消化系统常见恶性肿瘤，食管癌的高侵袭和转移特性是患者死亡的主要原因之一，最新研究发现肿瘤干细胞与肿瘤的浸润转移密切相关，本项目以与食管癌干细胞样细胞相关分子FNDC3B为研究对象，通过以下三方面研究结果，分析、阐明了miR34a/FNDC3B调控肿瘤干细胞样细胞干性影响食管癌浸润转移的机制。（1）食管癌组织中FNDC3B蛋白呈高表达，且与食管癌的浸润深度、淋巴结转移及总生存率密切相关，可以作为判断哈族食管鳞癌患者生存预后的关键因素；食管癌干性分子Nanog、BMI1和侵袭转移分子MMP2及FNDC3B浸润转移相关调控信号通路蛋白Akt在食管癌组织中均高表达，且FNDC3B与食管癌干性分子、MMP2及p-Akt蛋白表达水平正相关。（2）体外细胞学水平运用球体形成实验、质粒转染、Western blot、qRT-PCR、Transwell侵袭迁移实验等发现miR-34a及其靶基因FNDC3B影响食管癌干细胞样细胞的干性（如增殖能力，成球能力，球体分化能力，化疗敏感性等）及侵袭迁移能力。（3）机制研究显示miR-34a/FNDC3B通过PI3K/AKT/GSK-3β信号通路影响食管癌干细胞样细胞干性、克隆形成能力、侵袭迁移能力等，因此miR-34a/FNDC3B调节食管癌干细胞样细胞的干性特征及浸润转移，提示miR-34a或FNDC3B在食管癌干细胞的靶向治疗策略中具有一定的价值。