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Ras信号通过细胞外信号调节激酶(extracellular signal regulated kinase,ERK1/2)的磷酸化调节通路活性,Ras突变或者异常激活都会导致肿瘤发生。为了研究在MCF-7乳腺癌细胞中,激活Ras信号通路后组蛋白H4K16位发生乙酰化修饰状态(H4K16ac)水平对癌细胞增殖、迁移能力的影响,研究采用Western blot技术检测不同转染组的ERK1/2、p-ERK1/2、H4和H4K16ac的表达水平;免疫组织化学技术检测乳腺癌组织和癌旁组织中p-ERK1/2水平和H4K16ac水平;分别构建突变质粒激活细胞Ras信号通路和模拟细胞内H4K16ac;CCK-8技术检测细胞的增殖能力,Transwell技术检测细胞的迁移能力。在对照组,激活Ras信号组,模拟H4K16ac组中,p-ERK1/2相对表达水平分别为0.285±0.017、0.407±0.026、0.373±0.028;H4K16ac相对表达水平分别为0.331±0.013、0.082±0.005、0.082±0.007。在乳腺癌组织和癌旁组织中p-ERK1/2表达分别为0.064±0.001、0.051±0.001;H4K16ac表达分别为0.028±0.003、0.063±0.005。激活Ras信号通路后癌细胞的增殖和迁移能力分别增加34.8%和103.1%(p<0.05),与激活Ras信号通路的癌细胞相比,模拟H4K16ac的细胞的增殖和迁移能力分别降低25.1%和42.7%(p<0.05)。结果表明在MCF-7乳腺癌细胞中激活Ras信号通路后p-ERK1/2水平上升,导致H4K16ac降低。细胞中H4K16ac上升会抑制Ras信号通路活性,从而抑制乳腺癌细胞的增殖和迁移。H4K16ac在癌细胞的增殖和向周围组织转移过程中发挥重要作用。
Ras signaling regulates pathway activity through phosphorylation of extracellular signal regulated kinase (ERK1 / 2), Ras mutation or aberrant activation both resulting in tumorigenesis. In order to investigate the effect of H4K16ac on the proliferation and migration of cancer cells in the MCF-7 breast cancer cells, the expression of H4K16 at histone H4K16was activated by Ras signaling pathway. Western blot was used to detect the expression of H4K16ac in different transfection groups ERK1 / 2, p-ERK1 / 2, H4 and H4K16ac were detected by immunohistochemistry. The expression of p-ERK1 / 2 and H4K16ac in breast cancer tissues and paracancerous tissues were detected by immunohistochemistry. Simulated intracellular H4K16ac; CCK-8 technology to detect cell proliferation, Transwell technology to detect cell migration. In the control group, the relative expression levels of p-ERK1 / 2 in the Ras signaling group and the mock H4K16ac group were 0.285 ± 0.017, 0.407 ± 0.026 and 0.373 ± 0.028, respectively; the relative expression levels of H4K16ac were 0.331 ± 0.013 and 0.082 ± 0.005, 0.082 ± 0.007. The expression of p-ERK1 / 2 in breast cancer tissues and paracancerous tissues was 0.064 ± 0.001 and 0.051 ± 0.001, respectively. The expression of H4K16ac was 0.028 ± 0.003 and 0.063 ± 0.005, respectively. The proliferation and migration of cancer cells increased by 34.8% and 103.1% (p <0.05), respectively, after activation of Ras signaling pathway. Compared with cancer cells that activate Ras signaling pathway, the proliferation and migration ability of H4K16ac cells were reduced by 25.1% and 42.7% (p <0.05). The results show that the activation of Ras signaling pathway in MCF-7 breast cancer cells increased p-ERK1 / 2 levels, resulting in decreased H4K16ac. H4K16ac increase in cells will inhibit Ras signaling pathway activity, thereby inhibiting breast cancer cell proliferation and migration. H4K16ac plays an important role in cancer cell proliferation and metastasis to surrounding tissues.