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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。
OBJECTIVE: To observe the expression of MKK4 protein in breast cancer cell lines, investigate the effect of MKK4 protein expression on the motility and EMT markers of breast cancer cells, determine the role of MKK4 in the EMT transformation and tumor metastasis of tumor cells, and to study the mechanism of tumor metastasis Provide some basic information, lay a certain theoretical foundation for cancer prevention and treatment. METHODS: Cultured lysates of a series of breast cancer cell lines were collected by in vitro cell culture technique. Western blot was used to detect the expression of MKK4 and EMT markers in cell culture lysates and the corresponding maps of MKK4 expression and cell metastasis were established. siRNA technology to interfere with the MKK4 overexpression of breast cancer cell line MKK4 expression, Western blot technology to observe the MKK4 low expression of EMT markers changes, the same time, MKK4 plasmid was constructed transfected MKK4 low expression of breast cancer cell lines, Western blot observation Changes in EMT markers after MKK4 overexpression. MTT method, Transwell, scratch method were used to observe the changes of cell proliferation, motility and migration after MKK4 overexpression. Results: The expression of MKK4 protein in breast cancer cell lines was correlated with the motor ability of breast cancer cells, and was correlated with the expression of EMT markers. The higher the expression of MKK4 protein, the poorer the cell motility. The expression of MKT4 in breast cancer cell lines was affected by interference or transfection technology. The expression of MKT4 in breast cancer cell lines also changed accordingly. After MKK4 was overexpressed in breast cancer cell lines, the proliferation of MKK4 cells was significantly inhibited and the ability of migration and migration decreased accordingly. CONCLUSION: The expression of MKK4 protein in breast cancer cells is correlated with the EMT transformation and motor ability of breast cancer cells. The higher the MKK4 protein expression, the poorer the cell motility.