EVPsort Extracellular Vesicle and Particle profiling and sorting

Syndecan-1 controls lung tumorigenesis by regulating microRNAs packaged in exosomes [A549]

Abstract

We evaluated the effect of syndecan-1 in various cell-based and animal models of lung cancer and found that lung tumorigenesis was promoted when cells lose syndecan-1 expression. We also demonstrate that syndecan-1 (or lack thereof) alters the microRNA (miRNA) cargo carried within exosomes exported from lung cancer cells. Analysis of the changes in miRNA expression identifies a distinct shift toward augmented pro-cancerous signaling, which are consistent with the changes found in lung adenocarcinoma. In all, our work identifies syndecan-1 as an important factor on lung cancer cells that control its ability to shape the lung tumor microenvironment through alterations in miRNA packaging within exosomes. Overall design: Examiniation of exosomal microRNA profile in 2 sets of human non-small cell lung cancer cell lines (A549 cell line); The first set is scramble (wild-type; A549.Scrb) cells and syndecan-1 knock-down (A549.1735) cells

Extraction Protocol

Exosomes were isolated from cell culture media using using ExoQuick-TCTM (SBI), and microRNA was purified using QIAGEN miRNeasy micro kit (QIAGEN, Valencia, CA) per the manufacture’s protocols Libraries were prepared according to standard Ilumina's instructions

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