<i>Supporting data for </i><i>“</i>Functional characterization of low-density lipoprotein receptor-related protein 1B (LRP1B) and Mitotic arrest deficient 2 like 2 (MAD2L2) in esophageal squamous cell carcinoma (ESCC<b>)</b><i>”</i>

This study aimed to elucidate the underlying molecular pathogenesis of ESCC by investigating the functions of the two critical genes, <i>Mitotic arrest deficient 2 like 2</i> (<i>MAD2L2</i>) and <i>Low-density lipoprotein Receptor Related Protein 1B</i> (<i>LRP1B</i>), with frequent alterations in its development. lenti-CRISPR-Cas9 was used to depletion the expression of the genes. The depletion effects were examined by in vitro and in vivo subcutaneous tumorigenesis assay. RNA-seq was used to explore the downstream factors of the genes. RT-qPCR was used to validate the expression of the immune cell markers and the immune response genes. These experiments aim to examine the effect of the depletion of these two genes on ESCC cell lines and ESCC tumor microenvironment.本研究旨在阐明食管鳞状细胞癌（ESCC）的潜在分子发病机制，通过探究在该癌症发生发展过程中频发变异的两个关键基因——有丝分裂阻滞缺陷2样2（Mitotic arrest deficient 2 like 2，MAD2L2）与低密度脂蛋白受体相关蛋白1B（Low-density lipoprotein Receptor Related Protein 1B，LRP1B）的功能。本研究采用慢病毒CRISPR-Cas9系统敲低上述两个基因的表达，通过体外实验与体内皮下成瘤实验验证基因敲低效果；借助RNA测序（RNA-seq）技术探索这两个基因的下游调控靶点，并采用实时定量聚合酶链反应（RT-qPCR）验证免疫细胞标志物及免疫应答相关基因的表达水平。本系列实验旨在探究这两个基因的敲低对食管鳞状细胞癌细胞系及肿瘤微环境的影响。