Objective: To compare the difference of the target DNA binding sequences of Twistl in cells that overexpress Bcl-2 or not using chromatin immunoprecipitation. Methods: The hepatocellular carcinoma cell lines was screened and the cell line with normal twist expression was selected. Cells were transfected with Bcl-2 expression plasmid and control empty vector. After formaldehyde cross-linking, chromatin immunoprecipitation was performed to get the target DNA binding sequences of Twistl to be sequenced and analyzed. Results: After screening of 5 hepatocellular carcinoma cell lines, HepG2 was observed to express twist. There was a significant difference of the target DNA binding sequences of Twistl in cells that overexpress Bcl-2 or not. The amount of binding-DNA was 68 and 212, respectively. At the same time, they shared 154 sequences. Conclusion: There are 212 DNA binding sequences of Twistl in different mi-croenvironment, involving adhesion molecules, cytoskeleton, apoptosis, cell motility and migration and other biological functions.%目的:利用染色质免疫共沉淀测序研究Twist1结合的靶DNA序列,比较过表达Bcl-2和正常状态下Twist1结合靶DNA序列的变化.方法:筛选常态表达Twist1的肝癌细胞系,分别转染Bcl-2表达质粒和对照空载体,收集转染后细胞以甲醛交联DNA,收集样品进行染色质免疫共沉淀,获得与Twist1蛋白结合的DNA片段进行测序分析.结果:通过对5种肝癌细胞系进行筛选,观察到HepG2可表达Twist1,其与DNA结合序列在空载体对照组和Bcl-2过表达组具有很大的差别,结合DNA数量分别为68条和212条,其中共享序列154条.结论:Twist1在不同环境下与DNA结合位点多达212个,涉及粘附分子、细胞骨架、信号转导、代谢和转录调节等多种生物学功能.
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