This pathway responds to signals from a variety of growth factors (EGF, NGF, PDGF, etc.), mitogens and environmental stimulations, eventually leading to activation and phosphorylation of extracellular selleck compound signal-regulated kinase (ERK) through the signal amplification cascade. Phosphorylated ERK translocates to nucleus, where it acts on the AP-1, NF-κB and other nuclear transcription factors, thereby regulating
gene expression and promoting tumor cell proliferation, differentiation and survival. Over-activation of ERK has been found in many human malignant tumors including oral cancer, melanoma and breast cancer[2, 3]. Urinary trypsin inhibitor ulinastatin as a broad-spectrum protease inhibitor can inhibit trypsin, chymotrypsin, plasmin, human leukocyte elastase and hyaluronidase. It has anti-tumor metastasis and protective effects on patients accepted radiotherapy and chemotherapy and been widely used to treat acute pancreatitis and shock and to improve surgical outcome in clinic. Ulinastatin can bind to tumor cells through its N-terminal Selleck QNZ domain I
and exert its inhibitory effect on proteolytic activity of plasmin by binding to tumor cells through its C-terminal domain II, the major anti-fibrinolytic group. The impact of ulinastatin on uPA is more complicated. In addition to its inhibitory effects on gene transcription, it also inhibits uPA protein expression by affecting kinase C and MEK/ERK/c-Jun signaling pathways[4, 5]. To find a more effective treatment for breast cancer, this study check details explored PRKACG the additive effects of docetaxel and ulinastatin on the proliferation of breast cancer MDA-MB-231 cells and tumor growth in nude mice. Materials and methods 1. Materials Ulinastatin was purchased from Guangdong Techpool Bio-Pharma Co., Ltd. Docetaxel was bought from Sanofi-Aventis (French). SYBR Green/ROX qPCR Master Mix (2X) were purchased from Fermentas Inc. (Canada). Anti-uPA antibody was from Bioworld (USA). Anti-uPAR and anti-pERK antibodies were from Santa Cruz (USA). 24 well Transwell plates were from Corning (USA). Matrigel was from BD Company (USA). 2. Cell culture Human
breast cancer cell line MDA-MB-231 (ER-) and MCF-7 (ER+) were kindly gifted by Shanghai Institute of Biological Sciences, Chinese Academy of Sciences, and maintained in RPMI-1640 medium supplemented with 10% fetal bovine serum, 100 U/mL penicillin, 100 mg/L streptomycin at 37°C in an incubator supplemented with 5% CO2 under saturated humidity. 3. Animals 100 female BALB/c (nunu) mice at age 4-6 weeks and with body weight of 17-21 g from Animal Research Center of Chongqing Medical University (Production License No.: SCXK (Beijing) 2005-0013, the use permit number: SYX (Chongqing) 2007-0001) were kept in SPF-class environment at 22-25°C and 50-65% humidity. Drinking water, feed and experimental materials were sterilized and all experiments were complied with sterile principle. 4.