Real-time quantitative PCR analysis of human metastasis-related genes in cell pairs with different ROS stress conditions.

HEK293 T-Rex/K-ras cells were induced by doxycycline (20 ng/mL) for 6 h, K-ras/Off cells were treated with 15 mU GOx for 6 h, whereas K-ras/On cells were treated with 2 mM NAC for 6 h. Expression of metastasis-related genes was analyzed using the Human Tumor Metastasis RT² Profiler PCR Array. Human Tumor Metastasis RT² Profiler PCR Array profiling. HEK293 T-Rex/K-ras cells were used and treated with 15 mM GOX or 2 mM NAC separately as indicated in the summary. RNA was extracted from the cells using Trizol (T9424, Sigma), and cDNA was synthesized with the PrimeScript™ RT-PCR Kit (RR014B, Takara). Equal amount of total RNA extracted from cells with different pre-treatment was quantified prior to RT-PCR assay. Gene expression was analyzed by qPCR using TB Green® Fast qPCR Mix (RR430S, Takara) on a CFX384 Real-Time System (Biorad). Microarray analyses were performed according to the protocol for human tumor metastasis RT² Profiler PCR Arrays (PAHS-028Z, QIAGEN).