Effect of loss of ciliogenesis on myoblast quiescence

Quiescence is an actively maintained state of the cell cycle;aberrations in which can result in severe consequencces for development and tissue homeostasis. Primary cilia are signaling centres derived from the centrosome which are associated with quiescence.Using a mouse skeletal myoblast cell culture system that can be induced to exit the cell cycle reversibly into quiescence or irreversibly into differentiation, we studied the effect of loss of ciliogenesis on quiescence. We performed microarray analysisis to study changes in the global transcriptome of IFT88KD myoblasts in different cell cycle states. RNA was harvested from C2C12 myoblasts that underwent siRNA treatment using either IFT88 or negative control siRNA at four different cell cycle states- Proliferating (MB), Quiescent (G0), Reentry into cell cycle for 2 hours (R2), and 48 hours into terminal differentiation (D48). RNA was converted to cDNA, labelled and used to probe Affymetrix mouse 420A 2.0 arrays.

细胞静止（Quiescence）是细胞周期中受主动维持的稳定状态，该状态出现异常会对个体发育与组织稳态造成严重不良后果。初级纤毛（Primary cilia）是起源于中心体的信号转导中心，与细胞静止状态密切相关。本研究采用可被诱导可逆退出细胞周期进入细胞静止状态，或不可逆进入分化状态的小鼠骨骼肌成肌细胞培养体系，探究纤毛发生缺陷对细胞静止状态的影响。我们通过微阵列分析（microarray analysis），研究了不同细胞周期状态下IFT88敲低（IFT88KD）成肌细胞的全局转录组变化。实验材料取自经过IFT88小干扰RNA（siRNA）或阴性对照siRNA处理的C2C12成肌细胞，这些细胞分别处于四种不同的细胞周期状态：增殖期（MB，Proliferating）、细胞静止期（G0，Quiescent）、重新进入细胞周期2小时（R2）以及终末分化48小时（D48）。将提取的RNA反转录为互补DNA（cDNA），经标记后用于与Affymetrix小鼠420A 2.0基因芯片阵列进行杂交检测。