Project description:PIWI-interacting RNAs (piRNAs) are thought to silence transposon and gene expression during development. However, the roles of piRNAs in somatic tissues are largely unknown. Here we report the identification of 555 piRNAs in human lung bronchial epithelial (HBE) and non-small cell lung cancer (NSCLC) cell lines, including 295 that don’t exist in databases termed as piRNA-Like sncRNAs or piRNA-Ls. Distinctive piRNA/piRNA-L expression patterns are observed between HBE and NSCLC cells. piRNA-L-163 (piR-L-163), the top down-regulated piRNA-L in NSCLC cells, binds directly to phosphorylated ERM proteins (p-ERM), which is dependent on the central part of UUNNUUUNNUU motif in piR-L-163 and the RRRKPDT element in ERM, and. The piR-L-163/p-ERM interaction is critical for p-ERM’s binding capability to filamentous actin (F-actin) and ERM-binding phosphoprotein 50 (EBP50). Thus, piRNA/piRNA-L may play a regulatory role through direct interaction with proteins in physiological and pathophysiological conditions.
Project description:To elucidate potential roles of piRNA in lung cancer, we analyzed global piRNA expression profiles in 8 NSCLC and 3 HBE cell lines. RNA-seq results showed that >99% of the approximately 4.5 million reads were between 26 and 32 bases. Most of the piRNA loci are represented by two or more identical reads. We observed a total 555 expressed mature piRNAs, distributed among chromosomes and mitochondria with bias in chromosomes 1 and 6. 99% of the piRNAs are mapped to intergenic regions (64%) or introns (35%) and 1% to exons. Of the 555 mature piRNAs, 260 (47%) have been recorded in NCBI whereas 295 (53%) are novel. These results provide a comprehensive view of piRNA expressed in human bronchial epithelial cells and NSCLC cells. To get the purified mature piRNAs, small RNA (<200nt) was separated from total RNAs firstly, and then piRNA were purified in one nucleotide resolution gel. A unique 6 nt barcode for every cell line to distinguish each reads from each specific cell line, and the right size of small RNAs were used library for Illumina sequencing.
Project description:To elucidate potential roles of piRNA in lung cancer, we analyzed global piRNA expression profiles in 8 NSCLC and 3 HBE cell lines. RNA-seq results showed that >99% of the approximately 4.5 million reads were between 26 and 32 bases. Most of the piRNA loci are represented by two or more identical reads. We observed a total 555 expressed mature piRNAs, distributed among chromosomes and mitochondria with bias in chromosomes 1 and 6. 99% of the piRNAs are mapped to intergenic regions (64%) or introns (35%) and 1% to exons. Of the 555 mature piRNAs, 260 (47%) have been recorded in NCBI whereas 295 (53%) are novel. These results provide a comprehensive view of piRNA expressed in human bronchial epithelial cells and NSCLC cells.
Project description:we profiled small RNAs binding to phosphorylated Serine (p-Ser), Threonine (p-Thr) and Tyrosine (p-Tyr) residues of proteins in human lung bronchial epithelial (HBE) and lung squamous cell carcinoma (SCC) cells. A total of 1986 unique p-Proteins interacted piRNAs and piRNA-Likes were called.
Project description:Protein glycosylation plays a fundamental role in a multitude of biological processes, and the associated aberrant expression of glycoproteins in cancer has made them attractive targets as biomarkers and therapeutic targets. In this study, we examined differentially expressed glycoproteins in cell lines derived from three different states of lung tumorigenesis: an immortalized bronchial epithelial cell (HBE) line, a non-small cell lung cancer (NSCLC) cell line harboring an activation KRAS mutation and a NSCLC cell line harboring an EGFR activation deletion. Mutations in KRAS and EGFR are two common, distinct, non-overlapping genomic alterations in NSCLC. Using a Triple SILAC proteomic quantification strategy paired with hydrazide chemistry N-linked glycopeptide enrichment, we identified 118 quantifiable glycopeptides in the 3 cell lines derived from 82 glycoproteins. Proteomic profiling revealed that 27 (24%) of the glycopeptides overexpressed in both of the NSCLC cell lines with 6 of the glycopeptides overexpressed only in the EGFR mutant cells and 19 of the glycopeptides overexpressed only in the KRAS mutant cells.
Project description:Protein glycosylation plays a fundamental role in a multitude of biological processes, and the associated aberrant expression of glycoproteins in cancer has made them attractive targets as biomarkers and therapeutic targets. In this study, we examined differentially expressed glycoproteins in cell lines derived from three different states of lung tumorigenesis: an immortalized bronchial epithelial cell (HBE) line, a non-small cell lung cancer (NSCLC) cell line harboring an activation KRAS mutation and a NSCLC cell line harboring an EGFR activation deletion. Mutations in KRAS and EGFR are two common, distinct, non-overlapping genomic alterations in NSCLC. Using a Triple SILAC proteomic quantification strategy paired with hydrazide chemistry N-linked glycopeptide enrichment, we identified 118 quantifiable glycopeptides in the 3 cell lines derived from 82 glycoproteins. Proteomic profiling revealed that 27 (24%) of the glycopeptides overexpressed in both of the NSCLC cell lines with 6 of the glycopeptides overexpressed only in the EGFR mutant cells and 19 of the glycopeptides overexpressed only in the KRAS mutant cells.
Project description:Genome-wide DNA-methylation profiles of human lung cancer cell lines and normal lung cells were generated by Infinium bead chip technology DNA methylation patterns of over 480,000 CpG sites were analyzed in normal human bronchial epithelial cells (NHBEC) and three non small cell lung cancer cell lines (NSCLC: A427, A549 and H322) using bisulfite-based Illumina 450K BeadChip arrays
Project description:Genes differentially expressed among cells constituting an in vitro human lung carcinogenesis model consisting of normal, immortalized, transformed and tumorigenic bronchial epithelial cells were identified. The differentially expressed genes were then analyzed to determine their relevance to the gene expression patterns of clinical non-small cell lung cancer (NSCLC) samples as well as the clinical outcome of patients with this disease.
Project description:Exosomes are 30-100 nm sized membrane vesicles released by cells into extracellular space and mediate the intercellular communication via transfer of proteins and RNAs. To better understand the function of these microvesicles in lung carcinogenesis, we employed a Triple SILAC quantitative proteomic strategy to examine the differential protein abundance between exosomes derived from an immortalized normal bronchial epithelial cell line and two non-small cell lung cancer (NSCLC) cell lines harboring distinct activating mutations in the cell signaling molecules Kirsten rat sarcoma viral oncogene homolog (KRAS) or epidermal growth factor receptor (EGFR). We were able to quantify 727 exosomal proteins derived from the three cell lines. Proteins associated with signal transduction are enriched in NSCLC exosomes which are functionally active in regulating cell proliferation. The study is the first to investigate protein abundance differences in exosomes derived from NSCLC cells and their normal counterparts, and reveals the role of exosomes in NSCLC cancer progression, which may have clinical implications in biomarker development for patients with NSCLC.