Project description:The calcium-activated chloride channel ANO1 (TMEM16A) is overexpressed in colorectal cancer (CRC) and associated with poor prognosis, but its role in regulating metastatic plasticity is not fully understood. To elucidate the transcriptomic networks governed by ANO1, we performed RNA sequencing on SW620 CRC cells stably expressing sh-control or sh-ANO1. Total RNA was extracted using Trizol reagent. Sequencing libraries were constructed with the NEBNext® Ultra RNA Library Prep Kit for Illumina® and sequenced on an Illumina Novaseq platform, generating 125-150 bp paired-end reads. Bioinformatics analysis included read alignment with HISAT2 (v2.0.5) and differential expression analysis using DESeq2 (v1.20.0) with a threshold of |log2(Fold Change)| ≥ 1 and adjusted p-value < 0.05. Functional enrichment analysis for Gene Ontology (GO) and KEGG pathways was performed with clusterProfiler (v3.8.1). Gene Set Enrichment Analysis (GSEA) was conducted using gene sets from MSigDB. Transcriptomic profiling identified 1,894 differentially expressed genes (DEGs) upon ANO1 knockdown, with 1,523 downregulated and 371 upregulated. Downregulated genes included key EMT and signaling components (e.g., PIK3C2A, PIK3CA, KRAS, ZEB1). GO analysis revealed enrichment in processes related to genomic integrity and cell division. KEGG pathway analysis highlighted significant enrichment in “Homologous recombination” and “Platinum drug resistance”. A trend-level downregulation was observed for the “mTOR signaling pathway”. This dataset illustrates that ANO1 knockdown drives broad changes in a pro-metastatic transcriptome network and implicates mTOR signaling as a potential downstream mechanism, providing a resource for understanding how ANO1 promotes CRC progression.
Project description:MACC1 is a decisive driver of metastasis. In this data set the expression changes of shRNA mediated knock down is analyzed. We used microarrays to detail the global programme of gene expression changing upon MACC1 knock down.
Project description:In order to understand the single cell nature of SW620/SW620-TET2CD, we used the SW620/SW620-TET2CD clone from Day6/Day9/Day12/Day15 to start our single cell sequencing and analysis the diffference of single cell gene expression from different samples.