IKK in mesothelioma

perform gene expression silencing for both IKKs in AE17, AB2 and KPM1 cells using lentiviral shRNA pools. After validating gene silencing, RNA was extracted from both parental (shC) and daughter cells (shChuk, shIkbkb) and was send for RNAsequencing in order to define possible target genes, up or down regulated upon gene expression manipulation with lentiviral shRNA. These cell lines (parental and daughter) were evaluated in vitro for the role of IKKs in cell proliferation, survival and clonogenicity and in vivo for their impact in MPM development upon intrapleural injection in mice The purpose of this study was to functionally examine the main NF-κB activating kinases IKKα and ΙΚΚβ in mouse models of human and murine malignant pleural mesothelioma (MPM). First, we induced MPM in C57BL/6 and Balb/c mice by intrapleural injections of syngeneic AE17 and AB2 MPM cells respectively

采用慢病毒短发夹RNA（lentiviral shRNA）混合文库，对AE17、AB2及KPM1细胞中的IκB激酶（IKKs）实施基因沉默操作。在验证基因沉默效果后，分别从亲本细胞（shC对照组）与敲降细胞（shChuk、shIkbkb组）中提取总RNA，随后送检RNA测序（RNA sequencing），以明确经慢病毒shRNA介导的基因表达调控后，可能出现上调或下调的靶基因。本研究对上述亲本细胞与敲降细胞系开展体外实验，以探究IKKs在细胞增殖、存活及集落形成能力中的作用；同时通过向小鼠胸膜腔内注射细胞，开展体内实验以评估IKKs对恶性胸膜间皮瘤（malignant pleural mesothelioma, MPM）发生发展的影响。本研究的核心目的为，在人源及鼠源恶性胸膜间皮瘤（MPM）的小鼠模型中，对核因子κB（NF-κB）的主要激活激酶IKKα与IKKβ开展功能验证。首先，我们分别向C57BL/6及Balb/c小鼠胸膜腔内注射同源性AE17与AB2 MPM细胞，以此诱导构建MPM小鼠模型。