ON123300对ER阳性人乳腺癌MCF-7细胞的影响及机制研究

目的 初步探讨新型多靶点激酶抑制剂ON123300对ER阳性、HER2阴性的人乳腺癌MCF-7细胞增殖、周期、凋亡的影响及机制。方法 取处于对数生长期的MCF-7细胞,分为DMSO对照组、Palbocilib处理组及ON123300处理组,给药干预48 h后观察细胞生长状态,CCK-8法检测各组MCF-7细胞增殖抑制情况,流式细胞术(Flow cytometry,FCM)检测细胞周期和凋亡,Western blot检测cyclinD1、CDK4、pRb1、survivin和PI3K的表达。结果 光学显微镜下可见DMSO对照组MCF-7细胞贴壁良好,增殖迅速;Palbocilib处理组的MCF-7细胞增殖出现轻微抑制和少量细胞脱落,未见细胞形态改变;ON123300处理组MCF-7细胞增殖明显受抑制,数量明显减少,细胞形态改变,出现脱落现象。细胞抑制率随药物浓度增加而增加,呈现剂量依赖效应。ON123300处理组的MCF-7细胞明显阻滞在G₁/S期,并伴有细胞凋亡增加,差异有统计学意义(P＜0.05)。ON123300处理组cyclinD1、CDK4、pRb1、survivin、PI3K的蛋白表达量随药物浓度增高均逐渐降低,差异具有统计学意义(P＜0.05)。结论 ON123300对人乳腺癌细胞MCF-7有显著的增殖抑制、促进细胞周期阻滞和凋亡的作用,其机制可能与survivin/cyclinD1/CDK4/Rb1/和PI3K信号通路的抑制相关。

Effect of ON123300 on ER+ human breast cancer MCF-7 cells and its mechanism

Objective The objective of this study was to investigate the effect and mechanism of novel multi-target kinase inhibitor ON123300 on the proliferation,cycle and apoptosis of ER-positive and HER2-negative human breast cancer MCF-7 cells.Methods MCF-7 cells in logarithmic growth phase were treated with ON123300 or palbocilib for 48 h.DMSO was used as the control.The cellular morphology of MCF-7 cells was observed by optical microscope and the cell proliferation was measured by CCK8 assay.Flow cytometry(FCM)was used to detect cell cycle and apoptosis.Western blot was used to examine the expression of cyclinD1,CDK4,pRb1,survivin and PI3K protein in MCF-7 cells.Results No cell morphological changes of MCF-7 cells were observed in the control group.In the palbocilib group,MCF-7 cells showed slight inhibition of cell proliferation and a small amount of cell shedding.In the ON123300 group,cell proliferation was significantly inhibited and cell morphology changes,showing a shedding phenomenon.The inhibitory rate increased with increasing drug concentration,showing a dose-dependent manner.The cells in the ON123300 treated group were significantly arrested at the G1/S phase of cell cyde and were associated with increased apoptosis,with a statistically significant difference(P＜0.05).The protein expression of cyclinD1,CDK4,pRb1,survivin and PI3K in the ON123300-treated group were obviously reduced than those in the control group(P＜0.05).Conclusion ON123300 can significantly inhibit proliferation,promote cell cycle arrest and apoptosis of MCF-7 cells.The mechanism may be related to the inhibition of survivin/cyclinD1/CDK4/Rb1/PI3K signaling pathways.