Journal: Molecular Cancer
Article Title: The anti-erbB3 antibody MM-121/SAR256212 in combination with trastuzumab exerts potent antitumor activity against trastuzumab-resistant breast cancer cells
Figure Lengend Snippet: MM-121 enhances trastuzumab-mediated inactivation of Akt and growth inhibition in two erbB2+ breast cancer cell lines. A , SKBR3 and BT474 breast cancer cells were untreated or treated with either trastuzumab or MM-121 alone, or their combinations for 24 hrs. Cells were collected and subjected to western blot analyses of P-erbB2, erbB2, P-erbB3, erbB3, P-Akt, Akt, P-MAPK, MAPK, or β-actin. B , SKBR3 and BT474 cells were plated onto 96-well plates and incubated at 37°C with 5% CO2. After 24 hrs, the culture medium was replaced with 0.1 ml fresh medium containing 0.5% FBS or the same medium containing the indicated concentrations of trastuzumab in the absence (Trast) or presence (Trast + MM-121) of MM-121 (10 μg/ml) for another 72 hrs. The percentages of surviving cells from each cell line relative to controls, defined as 100% survival, were determined by reduction of MTS. Bars , SD. Data show a representative of three independent experiments.
Article Snippet: Antibodies used for western blots were as follows: erbB2 (EMD Chemicals, Inc., Gibbstown, NJ); erbB3 and P-erbB2 (Tyr1248) (LabVision Corp., Fremont, CA); P-erbB3 (Tyr1289), P-MAPK (Thr202/Tyr204), MAPK, P-Akt (Ser473), and Akt (Cell Signaling Technology, Inc., Beverly, MA); Cyclin D1 (M-20), E2F1 (KH95), and p27 kip1 (F-8) (Santa Cruz Biotechnology, Inc., Santa Cruz, CA); and β-actin (Sigma Co., St. Louis, MO).
Techniques: Inhibition, Western Blot, Incubation