LASSO-based screening for potential prognostic biomarkers associated with glioblastoma.

Background: Glioblastoma is the most common malignancy of the neuroepithelium, yet existing research on this tumor is limited. LASSO is an algorithm of selected feature coefficients by which genes associated with glioblastoma prognosis can be obtained. Methods: Glioblastoma-related data were selected from the Cancer Genome Atlas (TCGA) database, and information was obtained for 158 samples, including 153 cancer samples and five samples of paracancerous tissue. In addition, 2,642 normal samples were selected from the Genotype-Tissue Expression (GTEx) database. Whole-gene bulk survival analysis and differential expression analysis were performed on glioblastoma genes, and their intersections were taken. Finally, we determined which genes are associated with glioma prognosis. The STRING database was used to analyze the interaction network between genes, and the MCODE plugin under Cytoscape was used to identify the highest-scoring clusters. LASSO prognostic analysis was performed to identify the key genes. Gene expression validation allowed us to obtain genes with significant expression differences in glioblastoma cancer samples and paracancer samples, and glioblastoma independent prognostic factors could be derived by univariate and multivariate Cox analyses. GO functional enrichment analysis was performed, and the expression of the screened genes was detected using qRT-PCR. Results: Whole-gene bulk survival analysis of glioblastoma genes yielded 607 genes associated with glioblastoma prognosis, differential expression analysis yielded 8,801 genes, and the intersection of prognostic genes with differentially expressed genes (DEG) yielded 323 intersecting genes. PPI analysis of the intersecting genes revealed that the genes were significantly enriched in functions such as the formation of a pool of free 40S subunits and placenta development, and the highest-scoring clusters were obtained using the MCODE plug-in. Eight genes associated with glioblastoma prognosis were identified based on LASSO analysis: RPS10, RPS11, RPS19, RSL24D1, RPL39L, EIF3E, NUDT5, and RPF1. All eight genes were found to be highly expressed in the tumor by gene expression verification, and univariate and multivariate Cox analyses were performed on these eight genes to identify RPL39L and NUDT5 as two independent prognostic factors associated with glioblastoma. Both RPL39L and NUDT5 were highly expressed in glioblastoma cells. Conclusion: Two independent prognostic factors in glioblastoma, RPL39L and NUDT5, were identified.

Two alternative splicing variants of a sugar gustatory receptor modulate fecundity through different signalling pathways in the brown planthopper, Nilaparvata lugens.

Insect gustatory receptors play crucial roles in multiple physiological behaviours. Although the alternative splicing of some gustatory receptors has been observed in insect species, differences in their ligands and functions have rarely been reported. Here, we cloned NlGr10a and NlGr10b, two alternative splicing variants of a sugar gustatory receptor gene in the brown planthopper (BPH), Nilaparvata lugens (Stål), and found that their ligands were different by calcium imaging assay. The ligands of NlGr10a were fructose and cellobiose, and the ligand of NlGr10b was arabinose. Subsequently, the RNAi results showed that knockdown of both splicing variants decreased the number of eggs laid by BPH females, and the egg hatching rate after knockdown of NlGr10a was significantly lower than that after knockdown of NlGr10b. Furthermore, NlGr10a promoted the fecundity of BPH through the AMPK- and AKT-NlVg/NlVgR signalling pathways, whereas NlGr10b promoted the fecundity only through the AMPK- and AKT-NlVg signalling pathways. These findings broaden our understanding of the difference in the ligands and functions of alternative splicing variants of gustatory receptors in insects.

Identification of putative fecundity-related gustatory receptor genes in the brown planthopper Nilaparvata lugens.

BACKGROUND: The insect gustatory system plays a central role in the regulation of multiple physiological behaviors and the co-evolution between insects and their hosts. The gustatory receptors (Gr) are important to allow insects to sense their environment. It is critical to the selection of foods, mates and oviposition sites of insects. In this study, the Gr family genes of the brown planthopper (BPH) Nilaparvata lugens Stål (Hemiptera: Delphacidae) were identified and analyzed, and their potential relationship to the fecundity of BPH was explored by RNA interference (RNAi). RESULTS: We identified 32 putative Gr genes by analyzing transcriptome and genome data from BPH. Most of these Gr proteins have the typical structure of seven transmembrane domains. The BPH Gr genes (NlGrs) were expressed in virtually all tissues and stages, whilst higher transcript accumulations were found in adult stages and in the midguts of females. Based on the phylogenic analysis, we classified NlGrs into five potential categories, including 2 sugar receptors, 2 Gr43a-like receptors, 7 CO2 receptors, 5 bitter receptors and 13 NlGrs with unknown functions. Moreover, we found that 10 NlGrs have at least two alternative splicing variants, and obtained alternative splicing isoforms of 5 NlGrs. Finally, RNAi of 29 NlGrs showed that 27 of them are related to the transcript levels of two fecundity related genes vitellogenin and vitellogenin receptor. CONCLUSIONS: We found 32 Gr genes in BPH, among which at least 27 are required for normal expression of fecundity markers of this insect pest. These findings provide the basis for the functional study of Grs and the exploration of potential genes involved in the monophagous character of BPH.