Over-expressed estrogen receptor-alpha up-regulates hTNF-alpha gene expression and down-regulates beta-catenin signaling activity to induce the apoptosis and inhibit proliferation of LoVo colon cancer cells

摘要

Epidemiologic studies reported that the prevalence of hereditary non-polyposis colon cancer (HNPCC) in male is about 1.5-fold higher than that in female. Decreases in circulatory estrogen (E-2) have been reported to downregulate the expression of E-2 receptor (ER) and significantly increase the risk of colorectal cancer. Patients that received E-2 replacement therapy were found to have a reduction in the incidence of colon adenoma and carcinoma. Furthermore, significant decreases in the expression of ER have been found in colorectal cancer specimens. Evidences strongly suggest the protective roles of E-2 and ER against colorectal cancer. However, the mechanisms of ER alpha effects on colorectal cancer cells remained un-clear. LoVo cells were transient transfected to overexpress ER alpha, DNA fragmentation and the activated caspases measurements were performed to evaluate apoptotic effects. Western blotting was used to evaluate protein levels, and luciferase activity assay to measure the Htnf-a promoter activity. The results clearly demonstrated that overexpressed ER alpha with or without E-2 (10(-8) M) treatment could activate caspase -8, -9, and 3 and induce DNA fragmentation in LoVo cell. At the same time, overexpressed ER alpha plus E-2 significantly increases the expression and promoter activity of hTNF-alpha, and the DNA fragmentation effect induced by E-2 plus ER alpha were reduced by the addition of hTNF antibody (0.1 ng(ml). In addition, E-2 plus ER alpha significantly upregulated p21 and p27 levels and downregulated the beta-catenin and its target genes, cyclin D1 and Rb, which regulate the cell cycle and cell proliferation. The results indicate that E-2 plus overexpressed ER alpha induce LoVo cell apoptosis might mediate through the increase of hTNF-alpha gene expression, which in turn activate caspase-8, -9 and caspase-3 and lead to the DNA fragmentation and apoptosis. E-2 plus ER alpha also showed the downregulation of beta-catenin signalings implicating the suppression of proliferation and metastasis of colorectal cells. Efforts aiming at enhancing ER alpha expression and(or activity may be proved to be an alternative therapy against colorectal cancer.