Cloning and sequencing of qRT-PCR products The qRT-PCR products of lncRNA were first purified using a UNIQ-10 PCR Product Purification Kit and then cloned into the pUCm-T vector (Sangon Biotech, Shanghai, China) selleck following the manufacturer��s instructions. Then, DNA sequencing was performed by Sangon Biotech Co., Ltd. Serological tumor marker analysis Serum carcinoembryonic antigen (CEA) and carbohydrate antigen 19-9 (CA19-9) were measured using an Elecsys 2010 machine (Roche Diagnostics, Basel, Switzerland). The cutoff values were 5 ng/mL and 35 U/mL for CEA and CA19-9, respectively. Statistical analysis All statistical data were analyzed by Statistical Program for Social Sciences (SPSS) 18.0 software (SPSS, Chicago, IL) and GraphPad Prism 6.0 (GraphPad Software, La Jolla, CA).
One way analysis of variance test, two-tailed Student��s t-test and rank-sum test were used as appropriate. A receiver operating characteristic (ROC) curve was established to evaluate the diagnostic value. P<0.05 was considered statistically significant. Results LncRNA expression profiles in gastric cancer tissues relative to adjacent non-tumorous tissues The lncRNA expression patterns between gastric cancer tissues and adjacent non-tumorous tissues were found to be significantly different (Figure 1). Total of 135 lncRNAs, which expression change was more than twofold, were found (GEO accession numbers is 47850; http://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc="type":"entrez-geo","attrs":"text":"GSE47850","term_id":"47850"GSE47850). Among them, 71 and 64 were up and down expressed in tumor tissues, respectively.
The most down-regulated lncRNAs in gastric cancer tissues were FER1L4, uc001lsz, “type”:”entrez-nucleotide”,”attrs”:”text”:”BG491697″,”term_id”:”13453209″,”term_text”:”BG491697″BG491697, “type”:”entrez-nucleotide”,”attrs”:”text”:”AF131784″,”term_id”:”4406612″,”term_text”:”AF131784″AF131784, uc009ycs, “type”:”entrez-nucleotide”,”attrs”:”text”:”BG981369″,”term_id”:”14384104″,”term_text”:”BG981369″BG981369, “type”:”entrez-nucleotide”,”attrs”:”text”:”AF147447″,”term_id”:”4761798″,”term_text”:”AF147447″AF147447, HMlincRNA1600, and “type”:”entrez-nucleotide”,”attrs”:”text”:”AK054588″,”term_id”:”16549158″,”term_text”:”AK054588″AK054588.
The most up-regulated lncRNAs in gastric cancer tissues were H19, HMlincRNA717, “type”:”entrez-nucleotide”,”attrs”:”text”:”AI769947″,”term_id”:”5236456″,”term_text”:”AI769947″AI769947, Cilengitide “type”:”entrez-nucleotide”,”attrs”:”text”:”BQ213083″,”term_id”:”20393931″,”term_text”:”BQ213083″BQ213083, “type”:”entrez-nucleotide”,”attrs”:”text”:”AK054978″,”term_id”:”16549616″,”term_text”:”AK054978″AK054978, and “type”:”entrez-nucleotide”,”attrs”:”text”:”DB077273″,”term_id”:”83165446″,”term_text”:”DB077273″DB077273 (Table 1). Figure 1 Alterations in lncRNA expression profiles between gastric carcinoma tissues and non-tumorous tissues.