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

doi: 10.1186/1476-4598-12-134

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

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

doi: 10.1186/1476-4598-12-134

Figure Lengend Snippet: The addition of MM-121 enhances trastuzumab-induced cell cycle G1 arrest in erbB2+ breast cancer cell lines. SKBR3 and BT474 cells were untreated or treated with either trastuzumab or MM-121 alone, or their combinations for 24 hrs. A , Half of the cells were collected and subjected to western blot analyses with specific antibodies directed against E2F-1, Cyclin D1, p27 kip1 , or β-actin. The densitometry analyses of E2F-1, Cyclin D1, and p27 kip1 signals were shown underneath, and the arbitrary numbers indicate the intensities of each sample relative to controls, defined as 1.0. B & C , The other half of the cells were collected for analysis of cell cycle distributions by flow cytometry as described in the Materials and Methods. 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: Western Blot, Flow Cytometry

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

doi: 10.1186/1476-4598-12-134

Figure Lengend Snippet: MM-121 in combination with trastuzumab significantly inhibits in vivo growth of tumor xenografts established from BT474-HR20 trastuzumab-resistant breast cancer cells. BT474-HR20 cells were s.c. injected into nude mice to establish tumor xenografts. The tumor-bearing mice (n = 5) received i.p. injections of PBS, trastuzumab, MM-121, or trastuzumab plus MM-121 as described in the Materials and Methods. After 6 treatments, the mice were euthanized at day 36 post injection of tumor cells, and all tumors were excised for histology and IHC analysis. A , The graphs show the tumor growth curves. Bars , SD. The combinations of MM-121 and trastuzumab significantly inhibited tumor growth as compared to control or single Ab treatment. B , Data show the representative tumors with hematoxylin and eosin (H&E) staining and IHC analysis of erbB2 and erbB3. The residual tumor cells obtained from combinatorial treatments retained similar expression levels of erbB2/erbB3 receptors on the cell membrane.

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: In Vivo, Injection, Staining, Expressing