端粒酶逆转录酶（hTERT）通过NF-κB 抑制PLEKHA7促进肿瘤转移的分子机制研究

The relationship between telomerase reverse transcriptase (hTERT) and the metastasis of colon cancer has few studies. We previously found that the inhibition of PLEKHA7 can promote the metastasis of colon cancer cells, and the overexpression of hTERT can reduce PLEKHA7 expression. However, the mechanism of hTERT-mediate inhibition of PLEKHA7 is not yet known. We use Co-IP to confirm hTERT could bind p65, which is a NF-κB subunit, to form a protein complex. Then we use bioinformatics to find p65 could combine with PLEKHA7 promoter. These results suggest hTERT could bind p65 to inhibit PLEKHA7 promoter activity. In this study, we intend to use dual luciferase and chromosome immunoprecipitation (ChIP) to confirm hTERT / p65 complex can bind the promoter of PLEKHA7. By interfering with PLEKHA7 and overexpression of hTERT, we further study the important role of p65 and PLEKHA7 in hTERT-mediate metastasis of colon cancer in vitro and in vivo. This study intends to clarify hTERT/p65 can interaction with PLEKHA7 promoter to suppress PLEKHA7 promoter activity, thereby inhibiting PLEKHA7 expression and promoting the metastasis of colon cancer. The aim is to provide important theoretical basis for the treatment of colon cancer.

hTERT与结肠癌转移的关系少有研究.我们预实验发现抑制PLEKHA7可促结肠癌细胞转移,而hTERT可抑制PLEKHA7的表达,但其机制不清.我们通过Co-IP证实hTERT可与NF-κB 的p65亚基形成蛋白复合体,生物信息学预测到PLEKHA7启动子区域存在p65结合位点,提示hTERT抑制PLEKHA7表达的机制可能在于hTERT与p65形成复合物后作用于PLEKHA7的启动子,从而抑制PLEKHA7启动子活性.本研究拟采用双荧光素酶及ChIP等实验证实hTERT/p65复合体可作用于PLEKHA7启动子,通过干扰PLEKHA7及过表达hTERT,从体内外研究p65和PLEKHA7在hTERT促结肠癌转移中的作用.本研究拟阐明hTERT/p65可结合于PLEKHA7的启动子以抑制PLEKHA7启动子活性,下调PLEKHA7的表达,最终促结肠癌的转移.为结肠癌的治疗提供重要的理论基础