HAMNO (NSC111847)

Cell experiment:

Cell cycle assessment and γ-H2AX staining are monitored in UMSCC38 and OKF4 cells after 2 h incubation with HAMNO (2, 20, 50 μM) and fixed in 70% ethanol overnight. Cells are washed with PBS and incubated overnight in PBS containing 1% BSA, 10% goat serum and PS139-H2AX antibodies, washed and incubated in goat anti-mouse Alexa Fluor 647 antibody for 30 min at room temperature. Cells are incubated in 50 μg/mL propidium iodide and 100 μg/mL RNase A for 30 min, and 10,000 cells per sample are analyzed[1].

Animal experiment:

Athymic nude mice are used in this study. UMSCC38 and UMSCC11B cells are implanted into 6-week-old female mice by a single subcutaneous injection of tumor cells (2 to 6×105 cells in 100 mL of sterile PBS). The growth rates of tumors are determined by daily monitoring of tumor volume with vernier calipers [tumor volume=1/2(length×width2)]. Once the tumor size reaches 50 mm3, etoposide (10 mg/kg mouse) and HAMNO (2 mg/kg) are administered intraperitoneally every day for 3 days. Tumor size is monitored daily and the volume of the tumor is compared among all experimental groups. At least three mice are used per group[1].

References:

[1]. Glanzer JG, et al. RPA inhibition increases replication stress and suppresses tumor growth. Cancer Res. 2014 Sep 15;74(18):5165-72.