There was no significant difference in the distribution of demographic information between GSK2126458 datasheet patients enrolled and patients who did not. Written informed consent was obtained from each participant for the use of their DNA and clinical information. The study was approved by the ethnic committee of Qilu hospital. SNP genotyping Genomic DNA was extracted from 5-mL blood sample

that was collected from each patient upon recruitment. The OPN-66 T/G, -156(rs17524488), and −443(rs11730582) variants were genotyped by direct Selleckchem SRT1720 sequencing of the sense and anti-sense strands following polymerase chain reaction (PCR) amplification of the promoter regulatory region −473 to −3 (forward primer 50-CAA GCT ACT GCA TAC TCG AAA TCA CA-30; reverse YM155 supplier primer 50- ACA ACC AAG CCC TCC CAG AAT TTA-30), as previously described [19]. PCR was performed using 50 ng DNA as a template under the following conditions: 95°C for 10 min, then 36 cycles of 94°C for 30 s, an annealing temperature for 60 s, and 72°C for 60 s, with a final extension at 72°C for 15 min. After affinity membrane purification using the QIAquick Gel Extraction kit (Qiagen, Carlsbad, CA, USA), the PCR products were subjected to cycle sequencing with the respective forward and reverse primer using an automated ABI 3100 DNA sequencer by GeneCore Bio Technologies (Shanghai China). A 15% blind, random sample

of study subjects was genotyped twice by different persons (Yunzhen Chen and Haichun Liu) and the reproducibility was 100%. Statistical analysis Statistical

analysis was performed using SPSS 18.0 software. Quantitative variables departing from the normal distribution, including age, gender and smoking status were summarized. Comparison of age between cases and controls was assessed using an independent Student’s t-test. Comparison of gender, smoking stauts and genotype frequencies between cases and controls was assessed using a chi-square test and a Fisher’s exact test. Survival was calculated by the Kaplan-Meier method. All probability (P) values were two-tailed and statistical significance much was indicated as P < 0.05. Results Patient characteristics and clinical outcomes This study recruited 360 patients with lung cancer and 360 healthy controls. The baseline clinical characteristics of patients are summarized in Table 1. 79 patients (17.4%) were diagnosed with bone metastasis. By the time of the final analysis (July 2012), the median follow-up time had been 32 months. There were no significant differences in terms of distribution of age and gender, but significant on smoking status, suggest smoking is one of risk factors. Clinicopathologic characteristics of the patients and controls are shown in Table 1. Table 1 Clinicopathologic characteristics of patients with NSCLC and healthy controls   No. of patients or controls   Characteristics Case (n) Controls (n) P No. 360 360   Age, y     >0.05 Median 57.2 56.3   Range 24-81 23-87   Gender     >0.