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The original signal values were achieved by analyzing the chip gray scale scanning graph. Intensity for each feature of the array was determined as a single raw expression level for a gene. The screening criteria of differentially expressed genes should meet the following conditions simultaneously: (1) the different gene either in metastasis group http://www.selleckchem.com/products/carfilzomib-pr-171.html or in nonmetastasis group is an effective gene (P http://en.wikipedia.org/wiki/YES1 regulated genes and their corresponding GO terms was considered to be significant GO. A P value ��0.05 (Fisher's exact test) between the differentially regulated genes and their corresponding pathway was considered to be significant pathway. Differentially expressed genes, lipocalin 2 (LCN2) and PDZK1 interacting protein 1 (PDZK1IP1), were chosen to be confirmed by real-time quantitative reverse transcription http://www.selleckchem.com/products/azd9291.html PCR (real-time qRT-PCR) analysis. Primers were designed to have similar guanosine�Ccytosine content and melting temperatures (Table 3). Total RNA was extracted from 17 lung cancer tissues as mentioned previously in microarray experiment. Reactions were carried out with AB 7900 real-time quantization PCR apparatus (Applied Biosystems). Real-time qRT-PCR was performed in 10 ��L of final reaction volume with 1.0 ��L of RNA sample (50 ng/��L), 5 ��L of SYBR Green Real-time PCR Master Mix (Genepharma, Shanghai), and 2 ��L of primer. Negative control samples were run for each real-time qRT-PCR assay consisting of without RNA in the reverse transcriptase reaction and without cDNA in the PCR. Reactions were carried out under the following cycling conditions: 2 min at 50��C and 10 min at 95��C followed by 40 two-temperature cycles (15 sec at 95��C, 60 sec at 60��C), with the final stage being 15 sec at 95��C, 15 sec at 60��C, and 15 sec at 95��C. Fluorescence changes were monitored after each cycle, and melting curve analysis was done at the end of cycles to verify the identity of PCR product. Average cycle threshold (Ct) values were calculated from triplicate reactions. To adjust the different amounts of cDNA in each reaction, average Ct values for the target genes were normalized with an average Ct value of an endogenous control gene, ��-actin.