Project description:SOX2 is a transcription factor essential for self-renewal and pluripotency of embryonic stem cells. Recently SOX2 was found overexpressed in the majority of the lung squamous cell carcinoma (SQC), in which it acts as a lineage-survival oncogene. However, downstream targets/pathways of SOX2 in lung SQC cells remain to be identified. In order to identify genes/pathways likely to be downstream of SOX2, we conducted SOX2 silencing experiments in LK2 and NCI-H520 (H520 thereafter), two SOX2-abundant lung SQC cell lines and analyzed global gene transcription changes by gene expression microarray assay. Each of H520 and LK2 cell lines was treated with either pooled siRNAs of SOX2 or non-silencing (control) siRNAs. After 48 h, cells were harvested and totoal RNA extracted for gene expression microarray analysis using Illumina HumanHT12 v3 BeadChip.
Project description:SOX2 is a transcription factor essential for self-renewal and pluripotency of embryonic stem cells. Recently SOX2 was found overexpressed in the majority of the lung squamous cell carcinoma (SQC), in which it acts as a lineage-survival oncogene. However, downstream targets/pathways of SOX2 in lung SQC cells remain to be identified. In order to identify genes/pathways likely to be downstream of SOX2, we conducted SOX2 silencing experiments in LK2 and NCI-H520 (H520 thereafter), two SOX2-abundant lung SQC cell lines and analyzed global gene transcription changes by gene expression microarray assay.
Project description:The first and rate-limiting step in eukaryotic mRNA decay is the shortening of the poly(A) tail catalyzed by a family of enzymes known as deadenylases. In humans, several deadenylases have been recognized so far, yet it is not clear what the advantage is to have many enzymes catalyzing the same reaction. It is hypothesized that specific deadenylases may target unique subsets of mRNAs, or multiple deadenylases can act on the same mRNA, with discrete but overlapping functions. To understand the biological significance of the diversity of these enzymes we silenced the expression of several deadenylases, including PARN, NOC, CNOT6, CNOT6L, and CNOT7, in human cells of cancer origin (NCI-H520; squamous lung cancer), and analyze the impact on gene expression with microarrays.
Project description:Misplaced IgG expression in cancer cells has been implicated in exacerbated malignancy and poor clinical prognosis. Accumulating evidence indicate that a non-conventional sialylation modification is critical for the function of cancer-derived IgG, rendering the name sialylated cancer IgG (SIA-cIgG). However, our knowledge remains rudimentary about the functional role of SIA-cIgG in lung stemness maintenance. To investigate the mechanisms of SIA-cIgG in the regulation of lung stemness maintenance, we performed RNA-seq using established NCI-H520 cell lines sorted by FACS to have high or low expression of SIA-cIgG. This revealed that genes upregulated in SIA-cIgG high cells were associated with stem cell proliferation and maintenance.
Project description:Misplaced IgG expression in cancer cells has been implicated in exacerbated malignancy and poor clinical prognosis. Accumulating evidence indicate that a non-conventional sialylation modification is critical for the function of cancer-derived IgG, rendering the name sialylated cancer IgG (SIA-cIgG). However, our knowledge remains rudimentary about the regulatory mechanism that controls the expression and function of SIA-cIgG. To investigate the regulatory mechanisms of SIA-cIgG expression in lung cancer stem cells, we performed a genome-wide CRISPR activation screen in NCI-H520 cells by expressing a lentiviral library of 70,290 CRISPR-activating gRNAs targeting 23,430 genes. We investigated that the stemness-related transcriptional factors OCT4 (Gene name: POU5F1) and SOX2 were both among the top genes enriched in SIA-cIgG high cells, suggesting that OCT4 and SOX2 may serve as key factors promoting SIA-cIgG expression.
Project description:NFYC-AS1 is an overlapping antisense RNA transcribed head-to-head to NFYC sense gene, encoding for the subunit C of NF-Y transcription factor, which is known as master regulator of cell cycle and proliferation in normal and tumor cells. Here we performed NFYC-AS1 silencing in lung squamous carcinoma H520 cells by Gapmer antisense oligonucleotides and CRISPR/Cas9 TSS deletion. Afterwards, we performed differentially expressed analysis and gene set enrichement analysis to investigate on NFYC-AS1 function and mechanism of action.