ZBED2/STAT5A/ILK信号通路促进食管癌转移的功能机制及临床意义研究

Esophageal cancer is the sixth most common cancer and the fourth most frequent cause of cancer death in China, with esophageal squamous cell carcinoma (ESCC) being the predominant histologic subtype. The disease carries a dismal prognosis particularly for patients who present with metastatic esophageal cancer. Development of treatment strategies that can prevent the migration and invasion of cancer cells are therefore essential to tackle this highly aggressive disease. ..We have established (through in vivo selection) human ESCC cells (LM3) that are highly metastatic as cell line models that can be used to identify metastasis-associated microRNAs. The cDNA microarray profiling showed that ZBED2 was one of the most upregulated genes in the highly metastatic ESCC cells. The function of ZBED2 in esophageal cancer metastasis has not been documented previously. Our preliminary data showed that expression level of ZBED2 is significantly higher in ESCC tumor tissue compared with adjacent normal tissues, and deregulation of ZBED2 in ESCC is associated with patient survival and metastasis. In addition, expression level of ZBED2 was found to be further increased in metastatic ESCC in lymph nodes. Functional studies showed that forced expression of ZBED2 in ESCC cells robustly promoted cell invasion in vitro and tumor metastasis in vivo. Mechanistically, we found that ZBED2 could directly bind to promoter region and induce transcription of STAT5A (Signal transducers and activators of transcription), which was amongst the upregulated genes identified in ZBED2-overexpressing cells using RNA-seq, and was also predicted to be a transcription factor being able to bind to promoter of ILK. More importantly, knockdown of ILK or STAT5A could significantly abolished the promoting effects of ZBED2 on ESCC cell invasion. ..We therefore hypothesize that ZBED2 can promote tumor metastasis in ESCC through activation of STAT5A/ILK regulatory axis, and ZBED2 has its clinical significance in diagnosis and prognosis as novel biomarker and in precision medicine as therapeutic target. This regulatory mechanism has not been reported previously. In the proposed study, we will perform experiments with the following aims:.1..To study the effects of ZBED2 on ESCC cell invasion and metastasis.2..To investigate the molecular mechanisms how ZBED2 regulates ILK signaling pathway through activation of STAT5A transcription.3..To investigate the clinicopathological significance of ZBED2 expression in ESCC.4..To evaluate the therapeutic efficacy of ILK inhibitor in suppressing ESCC tumor metastasis in preclinical study...Deciphering the role of ZBED2 in activating STAT5A/ILK signaling pathway, and its function in promoting invasion and metastasis has great mechanistic and functional significance in the study of tumor metastasis. More importantly, the outcome of this project will facilitate the identification of effective biomarker in cancer diagnosis and prognosis, and provide useful preclinical data for development of novel therapeutic strategies for this lethal disease.

肿瘤转移是癌症治疗失败的主要原因，亟需深入研究和有效治疗方案。我研究组采用裸鼠尾静脉循环注射方法，建立高度转移食管癌细胞株，基因芯片分析筛选出ZBED2在食管癌高度转移细胞株中表达显著上调。然而，ZBED2在调控肿瘤转移方面的生物功能目前未知。我们发现，ZBED2在食管癌肿瘤组织中高表达，并且与患者生存期和转移显著相关，表达水平在转移瘤组织进一步升高。通过RNA测序、染色质免疫沉淀测序、生物信息学分析和一系列实验验证，我们提出科学假说：ZBED2可能直接结合STAT5A的启动子，通过增强STAT5A的表达水平而转录激活ILK信号通路，从而促进食管癌细胞侵袭和转移。我们拟从生物学功能、分子调控机制、潜在临床诊断意义、靶向治疗四个方面剖析ZBED2/STAT5A/ILK信号通路在肿瘤转移方面的重要作用。本项目将为肿瘤转移机制研究提供新线索，为临床提供新的无创诊断预后分子标记物和精准治疗靶点。

肿瘤转移是癌症治疗失败的主要原因，亟需深入研究和有效治疗方案。我研究组采用裸鼠尾静脉循环注射方法，建立高度转移食管癌细胞株，基因芯片分析筛选出ZBED2在食管癌高度转移细胞株中表达显著上调。我们发现，ZBED2在食管癌肿瘤组织中高表达，并且与患者生存期和转移显著相关，表达水平在转移瘤组织进一步升高。通过RNA测序、染色质免疫沉淀测序、生物信息学分析和一系列实验验证，我们发现ZBED2可能直接结合SOX9的启动子区域，通过增强SOX9的表达水平而转录激活ILK信号通路，从而促进食管癌细胞侵袭和转移。通过细胞水平和动物水平实验，本项目从生物学功能、分子调控机制、潜在临床诊断意义、靶向治疗四个方面剖析ZBED2/SOX9/ILK信号通路在肿瘤转移方面的重要作用，并通过分子对接虚拟筛选鉴定ILK抑制剂，评估其在抑制食管癌转移方面的治疗潜力。本项目为肿瘤转移机制研究提供了新线索，为临床提供了新的预后分子标记物和精准治疗靶点。