Regulation of gene expression by Ankrd35 in LKR10 cells grown in 2D monoculture or as 3D clusters in methylcellulose

The objective of this study was to use RNAseq to determine which gene change in expression when LKR10 cells are grown in monolayers (2D) or methylcellulose on ultra low attachment plates (3D), which prevents attachment of cells to the plate and promotes formation of 3D sphere-like clusters. A secondary objective was to determine whether knockdown of Ankrd35 altered gene expression of specific genes in 2D or 3D. Examination of gene expression in the murine LKR10 cell line with and without knockdown of Ankrd35 in traditional 2D monolayer cultures and in 3D methylcellulose cultures by RNAseq

本研究的首要目标为采用RNA测序（RNAseq）技术，探究LKR10细胞分别在单层贴壁培养（2D）以及置于超低吸附培养板的甲基纤维素中进行三维培养（3D）时发生表达变化的基因；其中3D培养体系可阻断细胞与培养板的贴附，进而促进三维球样簇的形成。本研究的次要目标为明确锚蛋白重复域35（Ankrd35）的基因敲低是否会改变2D或3D培养环境下特定基因的表达水平。本研究通过RNAseq技术，对分别经Ankrd35基因敲低处理与未处理的小鼠源LKR10细胞系，在传统2D单层培养及3D甲基纤维素培养中的基因表达情况进行了检测。