Genome-wide Evaluation Of Change In LncRNA Expression In Non-functioning Pituitary Adenoma

ObjectiveLong noncoding RNAs(lnc RNAs) have been implicated in the formation of many different types of tumors.The contributions of lnc RNAs to non-functioning pituitary adenomas(NFPAs) remain unknown. Here, we describe the expression profile of lnc RNAs in formalin-fixed and paraffin-embedded(FFPE) tissue specimens of human NFPAs and normal pituitaries(NPs) using microarray analyses and study their biology funtions preliniarily.Materials and MethodsTotal RNA from FFPEtissue specimens of five human NFPAs and five NPs was extracted, and synthetic c DNA was performed after the quality inspection qualified. Gene Chip? Human Transcriptome Array 2.0(HTA2.0) was used to describe the expression profile of lnc RNAs and m RNAs.The most differentially expressed 10 lnc RNAs were subsequently validated by quantitative real-time polymerase chain reaction(q RT-PCR). The functions of m RNAs were determined by pathway and Gene Ontology(GO) analyses, and the functions of lnc RNAs were identified by their co-expressing m RNAs.Results1. A total of 30020 lnc RNAs and 25994 m RNAs were detected in the two groups of FFPE samples. 87 lnc RNAs and 54 m RNAs were consistently up-regulated, while 26 lnc RNAs and 26 m RNAs were consistently down-regulated(fold-change ≥ 2.0, p < 0.05) in NFPAs, based on microarray analyses.2. Of the 113 candidate lnc RNAs identified by microarray analyses as potentially differentially expressed in NFPAs, we selected 10 lnc RNAs(n337303、n340496、n334406、n332607、n333074、n332409、 n334366、n335657、n409198 and MEG3) to validate the credibility of the microarrays. Results demonstrated that n337303, n340496, n334406, n332607, n333074, and n332409 were up-regulated and that n334366, n335657, n409198 and MEG3 were down-regulated, which were concordant with the microarray data.3. We found the highest enriched GOs targeted by up-regulated transcripts were “respiratory electron transport chain”(ontology: biological process), “vesicle”(ontology: cellular component) and “cytochrome-c oxidase activity”(ontology: molecular function). While the highest enriched GOs down-regulated transcripts were “peptide hormone processing”(ontology: biological process), “extracellular region”(ontology: cellular component), and “hormone activity”(ontology: molecular function). Furthermore, pathway analysis revealed that 99 pathways were correlated to the up-regulated transcripts, while 24 pathways were associated with the down-regulated transcripts.4. The lnc RNA-m RNA co-expression network predicted connections between the 10 lnc RNAs and their 57 coding genes(Pearson correlation coefficients(PCCs) no less than 0.9).Conclusion1. It is indeed effective that the expression profile of lnc RNAs in FFPE tissue specimens of human NFPAs and NPs performed by microarray analyses.2. We first systematically revealed the lnc RNA expression profile in NFPA using FFPE samples, and some of these lnc RNAs were validated by q RT-PCR to promote the credibility of the microarray results.3. Lnc RNAs may serve as biomarkers for human NFPAs.In addition, some of these lnc RNAs may constitute potential therapeutic targets of NFPAs.