ABSTRACT
OBJECTIVE: Previous studies have shown the carcinogenic role of long-chain non-coding RNAs (lncRNA) TRERNA1. However, the role of TRERNA1 in non-small cell lung cancer (NSCLC) has not been reported. This research aims to explore the regulatory effect of TRERNA1/FOXL1 axis on the malignant progression of NSCLC. PATIENTS AND METHODS: Quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) was performed to examine the expression levels of TRERNA1 and FOXL1 in 39 pairs of tumor tissues and paracancerous ones collected from NSCLC patients. The potential relation between TRERNA1 expression and clinical indicators of NSCLC patients was analyzed. Meanwhile, expression levels of TRERNA1 and FOXL1 in NSCLC cell lines were also detected by qRT-PCR. In addition, TRERNA1 knockdown model was constructed in H358 and SPC-A1 cells. Cell counting kit-8 (CCK-8), cell colony formation assay, and flow cytometry were applied to analyze the influence of TRERNA1 on NSCLC cell biological functions. Finally, Dual-Luciferase reporter gene assay and cell reverse recovery experiments were performed to figure out the underlying mechanisms of TRERNA1 in regulating NSCLC progression. RESULTS: QRT-PCR results indicated that the expression level of lncRNA TRERNA1 in tumor tissue samples of NSCLC patients was remarkably higher than that in adjacent tissues. Compared with NSCLC patients with low expression of TRERNA1, patients with high TRERNA1 expression had a worse pathological stage and overall survival. Similarly, compared with cells in sh-NC group, the proliferation ability of cells in sh-TRERNA1 group was remarkably attenuated. In addition, cell ratio in the G1 phase increased after knockdown of TRERNA1, suggesting the arrested G1/S cell cycle. Subsequently, FOXL1 was downregulated in NSCLC cell lines and tumor tissues. Meanwhile, FOXL1 level was verified to be negatively correlated with TRERNA1 level. Additionally, the binding between TRERNA1 and FOXL1 was confirmed by Dual-Luciferase reporter gene assay. Cell reverse investigation indicated the involvement of FOXL1 in TRERNA1-regulated malignant progression of NSCLC. CONCLUSIONS: LncRNA TRERNA1 was up-regulated both in NSCLC tissues and cell lines. Its level was associated with pathological stage and poor prognosis in NSCLC. In addition, lncRNA TRERNA1 could promote the malignant progression of NSCLC via modulating FOXL1.
Subject(s)
Carcinoma, Non-Small-Cell Lung/metabolism , Disease Progression , Forkhead Transcription Factors/biosynthesis , Lung Neoplasms/metabolism , RNA, Long Noncoding/biosynthesis , A549 Cells , Aged , Carcinoma, Non-Small-Cell Lung/genetics , Carcinoma, Non-Small-Cell Lung/pathology , Female , Forkhead Transcription Factors/antagonists & inhibitors , Forkhead Transcription Factors/genetics , Humans , Lung Neoplasms/genetics , Lung Neoplasms/pathology , Male , Middle Aged , RNA, Long Noncoding/geneticsABSTRACT
Retinitis pigmentosa (RP) is a retinal degenerative disorder that often causes complete blindness. Mutations of more than 50 genes have been identified as associated with RP, including the CACNA1F gene. In a recent study, by employing next-generation sequencing, we identified a novel mutation in the CACNA1F gene. In this study, we used the amplification refractory mutation system (ARMS) and identified a single nucleotide change c.1555C>T in exon 13 of the CACNA1F gene, leading to the substitution of arginine by tryptophan (p.R519W) in a Chinese individual affected by RP. This study actually confirms this novel mutation, and establishes the ARMS technique for the detection of mutations in RP.
Subject(s)
Base Pair Mismatch/genetics , Calcium Channels, L-Type/genetics , DNA Primers/metabolism , Mutation, Missense/genetics , Polymerase Chain Reaction/methods , Base Sequence , Heterozygote , Humans , Molecular Sequence Data , Point Mutation/genetics , Reproducibility of Results , Sequence Analysis, DNAABSTRACT
Study design:Disturbance of gastrointestinal motility following acute spinal cord injury complicated with paraplegia can lead to bacterial overgrowth in the gastrointestinal tract, and increase the incidence of bacterial translocation. Experiments in a New Zealand rabbit model of acute spinal cord injury were performed.Objective:This study was designed to determine if the electrical stimulation of the sacral nerve prevents gut-origin bacterial translocation and endotoxemia in an animal model of acute spinal cord injury.Settings:Xiangya Hospital, Central South University, Hunan, PR China.Methods:New Zealand rabbits were divided into three groups: Control group, Paraplegic Vehicle group without sacral nerve stimulation and Paraplegic Experiment group with sacral nerve stimulation. Blood and multiple organs were collected for bacterial cultures and endotoxin determination at 24, 48 and 72 h following spinal cord injury. The histology and ultra-structural features of the organs were studied.Results:Bacterial translocation and endotoxemia were observed in all animals with acute spinal cord injury. Sacral nerve stimulation increased defecation, decreased endotoxin levels and bacterial translocation and improved the morphology of the organs.Conclusion:After acute spinal cord injury, stimulation of the sacral nerve reduced gut bacterial translocation and endotoxemia.Spinal Cord advance online publication, 6 April 2010; doi:10.1038/sc.2010.35.
