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目的:观察SETD4(SET domain containing 4)蛋白在不同细胞中的表达和定位情况,以及亚砷酸钠(NaAsO2)刺激对细胞SETD4表达和定位的影响,探讨NaAsO2诱导SETD4的改变是否依赖于p38信号通路。方法:用Western blotting检测SETD4在不同细胞中蛋白的表达情况,用细胞免疫荧光技术检测SETD4在细胞中的定位;用NaAsO2刺激p38+/+和p38-/-细胞,分别收集细胞总蛋白检测SETD4蛋白的表达变化,同时观察SETD4的定位和移位改变。结果:SETD4蛋白无论在鼠源还是人源的细胞中均有表达;细胞免疫荧光结果显示SETD4在整个细胞中分布,以胞质较多;p38+/+和p38-/-细胞受NaAsO2刺激后其SETD4蛋白的表达趋势一致,即均在6 h时有明显减少;p38+/+细胞受NaAsO2刺激后SETD4蛋白从胞质向胞核移位,而p38-/-细胞SETD4的移位现象不明显。结论:SETD4蛋白在不同种属及组织来源的细胞中均有表达,表现为全细胞分布,以胞质为主;NaAsO2刺激可影响细胞SETD4蛋白的表达水平及定位,NaAsO2诱导的SETD4移位改变有赖于p38 MAPK介导的信号通路。
OBJECTIVE: To investigate the expression and localization of SET domain containing 4 (SETD4) proteins in different cells and the effect of sodium arsenite (NaAsO2) on the expression and localization of SETD4 in cells. To investigate whether the change of SETD4 induced by NaAsO2 depends on the expression of p38 path. Methods: The expression of SETD4 protein in different cells was detected by Western blotting. The SETD4 protein was detected by immunofluorescence staining in cells. The p38 + / + and p38 - / - cells were stimulated by NaAsO2. The changes of the expression of SETD4 were also observed. Results: SETD4 protein was expressed both in murine and human cells. Immunofluorescence results showed that SETD4 was distributed in the whole cell with more cytoplasm. When p38 + / + and p38 - / - cells were stimulated by NaAsO2 SETD4protein expression trends are consistent, which were significantly reduced at 6 h; p38 + / + cells after stimulation by SETS4 SETD4protein from the cytoplasm to the nucleus, while p38 - / - cells SETD4shift phenomenon is not obvious. CONCLUSIONS: SETD4 protein is expressed in all kinds of cells of different species and tissues. The distribution of SETD4 protein is cytoplasm. NaAsO2 stimulates the expression of SETD4 protein and the localization of SETD4 protein. The change of SETD4 translocation induced by NaAsO2 Depends on the p38 MAPK-mediated signaling pathway.