The effects of treatments with 2-methoxyestradiol, NanI and hNit1 in human embryonic kidney HEK293T cells

• Main Authors: Li-Ju Chen, 陳麗如
• Other Authors: Chin-Hsiang Chien
• Format: Others
• Language: zh-TW
• Published: 2013
• Online Access: http://ndltd.ncl.edu.tw/handle/54453882957714882950

碩士 === 國立陽明大學 === 生化暨分子生物研究所 === 101 === According to the vital statistics from Department of Health, malignant tumors listed in the first rank of 10 major death caused reasons in 2012. Traditional chemotherapy agents not only destroy tumor cells but also harm to normal cells and give rise to severe side-effects, drug-resistance of chemotherapy agents and problems with tumor metastasis also affect effectiveness and prognosis in patients. Therefore, many researchers are devoted to finding or developing more effective treatment. In this study, I try to investigate the effects of three different treatments drug, NanI and Pro1 in cancer cells for future translational research. Drug, a nature metabolite of 17β-estradiol, shows antiproliferative and antitumor properties in vitro and in vivo. I found that drug inhibited cell growth and caused G2/M phase arrest. NanI, a large sialidase from Clostridium perfringens, has been reported that NanI hydrolyzed the terminal sialic acids of glycoconjugates on human ovarian cancer cell line OVCAR-3 cell, suppressed the invasiveness and induced apoptosis. In this study, the treatment of NanI in cancer cells inhibited cell growth and induced apoptosis. More experiments are needed to investigate the mechanisms of NanI in cancer cell. Pro1 belongs to the nitrilase superfamily, previous studies reported that Pro1-deficient mouse kidney cells may accelerate cell proliferation and increase cyclin D1 expression. Pro1 overexpression induced caspase-dependent apoptosis in double knockout Pro1 mouse kidney cell. Previous studies in our Lab demonstrated that overexpression of Pro1 in cancer cells suppressed cell growth. In this study, I try to search the functional domain in Pro1 to determine the domain of Pro1. First of all, I want to know whether the N-terminal of 1-30 amino acids (Pro1 1-30) alone could inhibit cell growth. Overexpression of Pro1 significantly reduced the growth rate of cancer cells, but Pro1 1-30 did not. In addition, overexpression of Pro1 did not alter the progression of cell cycle but induced cell death. Taken together, the three independent treatments of drug, NanI and Pro1 showed different growth inhibition effects and mechanisms, the results may provide knowledge for animal experiments.