Bex1抑制结肠癌肝转移的功能、机制及转化研究

项目名称： Bex1抑制结肠癌肝转移的功能、机制及转化研究

项目编号： No.81472664

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 医药、卫生

项目作者： 丁克峰

作者单位： 浙江大学

项目金额： 64万元

中文摘要： 肝转移是导致结肠癌患者死亡的重要原因，但目前对肝转移的分子机制尚不明确。我们前期研究发现，与非转移性结肠癌组织相比，一个小分子蛋白Bex1在转移性组织中显著沉默。原代结肠癌肝转移细胞系中Bex1的表达也明显低于原代盲肠原位肿瘤细胞。动物模型也证实Bex1表达抑制后结肠癌细胞肝转移率明显增加。我们推测：Bex1表达可抑制结肠癌肝转移。本研究拟构建Bex1完全敲除结肠癌细胞系，应用细胞迁移侵袭实验及三维培养等体外实验平台，盲肠原位肿瘤肝转移等动物模型，评估Bex1被完全敲除后细胞肝转移能力；进一步运用基因芯片、定量PCR、免疫组化等技术筛选Bex1下游靶基因；用shRNA抑制上调的靶基因或过表达下调的靶基因后，在细胞和动物模型中验证可否逆转转移，最终明确Bex1在结肠癌肝转移中的功能和作用机制。我们也将探索Bex1靶基因抗体/抑制剂预防/治疗肝转移的可行性，为结肠癌肝转移的综合治疗探索新靶点。

中文关键词： Bex1基因；结肠癌；肝转移；分子机制；转化研究

英文摘要： Liver metastasis accounts for much of the colon cancer mortality. What makes it challenging to address this issue is that the molecular mechanism of metastasis is not fully understood. Our previous study found that a small molecular protein Bex1 was significantly silenced in metastatic colon cancer tissues, compared to non-metastatic colon cancer tissues. Bex1 expression was decreased statistically in hepatic metastatic cells versus its primary tumor cells, Which were all originated from the orthotopic colon cancer liver metastasis animal model. Further investigation also confirmed that suppressed expression of Bex1 in colon cancer cells promotes liver metastasis in subcutaneous tumor model. Bex1 was supposed to be a suppressor of liver metastasis in colon cancer based upon these studies. To fully understand of the function and mechanism of Bex1 plays in liver metastasis of colon cancer, we aim to evaluate the ability of liver metastasis of Bex1-knockout colon cancer cell line in vitro and in vivo. Specifically, we will use several kinds of methods including Transwell assay, Wound healing assay, 3-D culture, etc for in vitro study, and animal model such as orthotopic colon cancer liver metastasis mouse model for in vivo study. Multiple molecular biology techniques including gene microarray, Real time PCR, Immunohistochemistry, etc will be utilized to screen the downstream target genes of Bex1 inhibited liver metastasis. We plan to explore the possibility of reversing liver metastasis through inhibiting the expression of the up-regulated target genes with shRNA or over-expressing the down-regulated target genes by in vitro and in vivo tests, respectively. The role and molecular mechanism of Bex1 inhibited liver metastasis of colon cancer will be confirmed eventually. The feasibility of using antibody or inhibitor against Bex1 targets for preventing or treating liver metastasis will also be explored. The elucidation of Bex1's function and mechanisms could lead new insights into the comprehensive therapy of colorectal liver metastases.

英文关键词： Bex1 gene;Colon cancer;Liver metastasis;Molecular mechanism;Translational research