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目的探讨核因子κB信号通路抑制剂BAY11-7082通过调控三磷酸腺苷柠檬酸裂解酶(ACL)对MCF-7乳腺癌细胞增殖及凋亡的影响。方法选取对数生长期的MCF-7细胞,随机分为对照组和5μmol/L BAY11-7082处理组、10μmol/L LY294002处理组。采用Western blot法检测BAY11-7082和LY294002处理后MCF-7细胞中ACL、磷酸化的ACL(p-ACL)、磷酸化的Akt(p-Akt)、磷酸化的核因子κB(p-NF-κB)的蛋白水平。采用BAY11-7082和小干扰RNA敲低ACL(si ACL),CCK-8法检测MCF-7细胞的增殖、异硫氰酸荧光素标记的膜联素Ⅴ/碘化丙啶(annexinⅤ-FITC/PI)双标记结合流式细胞术检测MCF-7细胞的凋亡。结果 BAY11-7082能抑制MCF-7细胞增殖,且呈剂量依赖性。Werstern blot结果显示BAY11-7082处理MCF-7细胞48 h后,p-NF-κB和p-ACL明显下降;LY294002处理组中,p-Akt和p-ACL明显下降。CCK-8实验和流式细胞术检测结果显示,BAY11-7082和si ACL分别处理MCF-7细胞48 h后,细胞增殖减少,细胞凋亡明显增加。结论 BAY11-7082能通过抑制ACL的磷酸化从而抑制乳腺癌MCF-7细胞的增殖并促进其凋亡。
Objective To investigate the effects of BAY11-7082, an inhibitor of nuclear factor kappa B signaling pathway, on the proliferation and apoptosis of MCF-7 breast cancer cells through the regulation of adenosine triphosphate citrate lyase (ACL). Methods The logarithmic growth phase of MCF-7 cells were randomly divided into control group and 5μmol / L BAY11-7082 treatment group and 10μmol / L LY294002 treatment group. Western blot was used to detect the expression of ACL, p-ACL, p-Akt, p-NF- κB) protein levels. ACL (si ACL) was knocked down by BAY11-7082 and small interfering RNA. The proliferation of MCF-7 cells was detected by CCK-8 assay. The expression of annexinⅤ-FITC / PI) double labeling combined with flow cytometry detection of apoptosis in MCF-7 cells. Results BAY11-7082 inhibited the proliferation of MCF-7 cells in a dose-dependent manner. The results of Werstern blot showed that the expression of p-NF-κB and p-ACL significantly decreased after treated with BAY11-7082 for 48 h, while the expression of p-Akt and p-ACL decreased significantly in LY294002 group. The results of CCK-8 assay and flow cytometry showed that the proliferation of MCF-7 cells was decreased and the cell apoptosis was significantly increased after treated with BAY11-7082 and si ACL respectively for 48 h. Conclusion BAY11-7082 can inhibit the proliferation of breast cancer MCF-7 cells and promote its apoptosis by inhibiting the phosphorylation of ACL.