Influence of Recombination C1-Inhibitor on proliferation and migration of Human Breast Cancer Cells

• Main Authors: Chi-xin Ye, 葉慈馨
• Other Authors: Shiping He
• Format: Others
• Language: en_US
• Published: 2010
• Online Access: http://ndltd.ncl.edu.tw/handle/23456312399292969892

碩士 === 國立中山大學 === 生物科學系研究所 === 98 === Human C1 inhibitor (C1 INH) belongs to the superfamily of serine protease inhibitors (serpins). It is the only known physiological inhibitor of C1s and C1r, therefore C1 INH plays an essential role as a regulator of complement classical pathway. C1 INH is also one of the major inhibitors of plasma kallikrein and activated factor XII of the kinin system. There is evidence that C1 INH can also inhibit the complement lectin pathway proteases, and the complement alternative pathway (C3b), plasminogen activator, activated factor XI, and kallikrein. A recent report indicated C1 inhibitor was significantly down-regulated in normal breast cells compared to malignant cell line. Thus, the major aim of this project is to study the effect of recombinant C1 inhibitor Ile379-Ala478 on human breast cancer cells. In this study, we constructed plasmid containing DNA fragment for C1 inhibitor C-terminal negion of Ile379-Ala478. Then the constructed plasmid was transferred into E.coli BL21(DE3) for expression. The recombinant protein was purifed by GST-affinity chromatography. Recombinant C1 inhibitor produced by E.coli was used to treat breast cancer cells (MDA MB 435s) to study its effect on cell proliferation and migration. Results showed that recombinant C1 inhibitor Ile379-Ala478 protein inhibited proliferation of MDA MB 435s cells after four, six or eight days, incubation in the presence of recombinant protein (1~100 nM). However, cell migration was inhibited only at 100 nM on 48 hrurs incubation..Our experimental data showed that the recombinant C1 inhibitor Ile379-Ala478 inhibited proliferation and migration of MDA MB 435s cells.