Products/Services Used | Details | Operation |
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Recombinant Proteins> | The MCF-7, a human breast adenocarcinoma cell line, was obtained from Chinese academy sciences (Shanghai, China). Earthworms, E. foetida were provided by Huaneng Pharmaceutical Co. Ltd (Shandong, China). Columns of diethylaminoethanol (DEAE)-Sepharose FF, Sephacryl S-200 HR and Sephadex G-10 were purchased from GE (USA). Primers for glyceraldehyde 3-phosphate dehydrogenase (GAPDH), focal adhesion kinase (FAK) and CD44v6 were purchased from GenScript (Jiangsu, China); antibodies for GAPDH and FAK were purchased from Santa Cruz (USA). Dulbecco's modified Eagle medium (DMEM), foetal bovine serum (FBS) and trypsin were all purchased from Gibco (USA); CCK-8 kit Beyotime (Shanghai, China); human fibrinogen and thrombin were purchased from Sigma (USA). Other reagents and chemicals used were of analytical grade and commercially available. | Get A Quote |
Earthworm fibrinolytic enzyme from Chinese earthworm Eisenia foetida was isolated to investigate its antitumor activity in breast cancer cells. The protein isolated was characterised as earthworm fibrinolytic enzyme using the fibrin plate method. The molecular weight of earthworm fibrinolytic enzyme component was determined to be 25 kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis assay using a standard protein ladder. Earthworm fibrinolytic enzyme can markedly inhibit the growth and migration of MCF-7 cells in a dose and time dependent manner. In addition, MCF-7 cells treated with earthworm fibrinolytic enzyme (40 μg/ml) began to undergo apoptosis after 24 h. Expression of focal adhesion kin... More