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为了研究WSTF调控Ras相关乳腺癌细胞特性的机制。研究采用Western Blot检测WSTF磷酸化和组蛋白修饰水平;经GST pull-down验证WSTF与PCAF、MOF之间的相互作用,采用乙酰化酶活性实验验证PCAF和MOF酶活性;经Ch IP、Real-time PCR检测基因表达,并采用体内成瘤实验验证细胞成瘤能力。研究发现WSTF与PCAF、MOF之间存在相互作用。Ras通路激活后,WSTF磷酸化水平上调,使其与PCAF的结合增强的同时与MOF的结合减弱。这一变化引起PCAF的乙酰化酶活性上调而MOF的酶活性下调,导致H3K9ac上调和H4K16ac下调。组蛋白修饰的变化引起通路下游肿瘤相关基因表达发生改变,增强细胞成瘤能力。综上,WSTF蛋白通过调节与PCAF和MOF的相互作用,影响下游H3K9ac和H4K16ac水平及肿瘤相关基因表达,调控乳腺癌细胞成瘤能力。
To investigate the mechanism by which WSTF regulates Ras-related breast cancer cell characteristics. Western Blot was used to detect the phosphorylation and histone modifications of WSTF. The interaction between WSTF and PCAF and MOF was verified by GST pull-down. The activity of PCAF and MOF was verified by the activity of acetylase. time PCR to detect gene expression, and using in vivo tumorigenicity experiment to verify the ability of tumorigenesis. The study found that there is interaction between WSTF and PCAF and MOF. Activation of the Ras pathway leads to an upregulation of WSTF phosphorylation, resulting in a stronger binding to PCAF and a weaker binding to MOF. This change causes an up-regulation of acetylase activity of PCAF and a down-regulation of the enzymatic activity of MOF, resulting in up-regulation of H3K9ac and down-regulation of H4K16ac. Changes in histone modifications cause changes in the expression of tumor-associated genes downstream of the pathway, enhancing the ability of the cells to become tumorigenic. In summary, WSTF protein regulates the tumorigenicity of breast cancer cells by regulating the interaction with PCAF and MOF, affecting the levels of downstream H3K9ac and H4K16ac and tumor-related gene expression.