SW480 cells undergoes postmitotic neuron-like differentiation in response to EZH2 knockdown. SW480 cells undergoes postmitotic neuron-like differentiation in response to EZH2 knockdown

Formerly we found that combined inhibition of class I HDACs, DNMTs, EZH2 and LSD1 with chemical inhibitors in a variety of cancer cells caused neuronal differentiation. Our recent study demonstrated that functional knockdown of EZH2 alone in SW480 cells can also lead to significant neuronal differentiation. We used microarray to analyze global gene expression change in SW480 cells after neuronal differentiation induced by EZH2 knockdown. Overall design: EZH2 function was blocked in SW480 cells via shRNA knockdown. Cells underwent significant morphological change after 6 days of EZH2 knockdown, as compared control SW480 cells that was infected with lentivirus carrying empty vector. Control and EZH2 knockdown cells were then collected and subjected to microassay analysis. Two samples, one was control SW480 cells and the other was SW480 cells with EZH2 knockdown, were used for microarray assay.

本课题组此前的研究显示，在多种癌细胞中联合应用化学抑制剂抑制I类组蛋白去乙酰化酶（class I HDACs）、DNA甲基转移酶（DNMTs）、EZH2及赖氨酸特异性去甲基化酶1（LSD1），可诱导神经元分化。我们近期的研究证实，仅在SW480细胞中对EZH2进行功能敲低，即可引发显著的神经元分化。为分析EZH2敲低诱导神经元分化后SW480细胞的全局基因表达变化，我们采用微阵列（microarray）技术开展了检测。实验设计概述：通过短发夹RNA（shRNA）介导的敲低实验阻断SW480细胞中的EZH2功能。与感染携带空载体慢病毒的对照SW480细胞相比，EZH2敲低6天后的细胞出现了显著的形态学改变。随后收集对照细胞与EZH2敲低细胞，进行微阵列分析。本次微阵列实验共使用两个样本：其一为对照SW480细胞，其二为经EZH2敲低的SW480细胞。