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目的：观察乳腺癌细胞株中MKK4 蛋白的表达水平，研究MKK4 蛋白表达对乳腺癌细胞运动能力及EMT 标志物的影响，确定MKK4 在肿瘤细胞EMT 转化及肿瘤转移中的作用，为肿瘤转移机制研究提供一定的基础资料，为肿瘤防治奠定一定的理论基础。方法：通过体外细胞培养技术收集系列乳腺癌细胞株的培养裂解液，利用Western blot 技术检测细胞培养裂解液中MKK4 及EMT 标志物的表达水平，构建MKK4 表达水平与细胞转移能力的对应图；采用siRNA 技术，干扰MKK4 高表达乳腺癌细胞株MKK4 的表达，Western blot 技术观察MKK4 低表达后，EMT 标志物的变化，同时，构建MKK4 质粒，转染MKK4 低表达乳腺癌细胞株，Western blot 技术观察MKK4 高表达后，EMT 标志物的变化。并采用MTT 法、Transwell、划痕法观察MKK4 高表达后细胞增殖、运动、迁移等能力的变化。结果：乳腺癌细胞株中MKK4 蛋白的表达水平与乳腺癌细胞运动能力有一定的相关性，并与EMT 标志物的表达具有相关性，MKK4 蛋白表达越高，细胞运动能力越差。干扰或转染技术影响乳腺癌细胞株中MKK4 的表达后，细胞EMT 标志物的表达也相应变化，乳腺癌细胞株中MKK4 高表达后，其细胞增殖明显抑制，运动迁移能力也相应下降。结论：乳腺癌细胞中MKK4 蛋白的表达水平与乳腺癌细胞EMT 转化及运动能力有一定的相关性，MKK4 蛋白表达越高，细胞运动能力越差。

英文摘要:

Objective: To observe MKK4 protein expression level of MKK4 and its role in tumor cell transformation and tumor metastasis and to provide some basic data for cancer prevention and control. Methods: In vitro cell culture technology was employed to collect series of lysate of breast cancer cell lines. Western blot was used to detect the expression level of MKK4 and EMT markers. Structure the corresponding chart for MKK4 expression levels and transforming abilities. siRNA technology was used to interfere MKK4 expression in MKK4 high expression breast cancer cell lines. Western blot technique was then used to observe EMT markers when MKK4 expressed low. At the same time, built MKK4 plasmid then transfected MKK4 into breast cancer cell lines with low expression, and observed EMT changes when MKK4 expressed high. And MTT method, Transwell, and wound healing assay were used to observe capacity changes of cell proliferation, movement and migration. Results: MKK4 protein expression level was correlated with the breast cancer cell motility and the expression of EMT markers. The higher MKK4 expressed, the worse the cell movement capability was. EMT markers showed corresponding changes in breast cancer cells when interference or transfection influenced expression of MKK4 in breast cancer cell lines. With high expression of MKK4, cell proliferation was significantly inhibited and the migration capability was correspondingly reduced. Conclusion: There was a correlation between the protein expression level of MKK4 in breast cancer cell lines and the transformation and movement capability of EMT. The higher the expression of MKK4, the worse the moving capability of cells.

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