FGFR1抑制剂Ponatinib对乳腺癌细胞增殖凋亡的影响  

The influence of FGFR1 inhibitor Ponatinib on celluar proliferation and apoptosis of breast cancer cells

作　　者：王云 韩正祥 

机构地区：[1]徐州医科大学研究生学院,江苏徐州221000 [2]徐州医科大学附属医院肿瘤内科,江苏徐州221000

出　　处：《现代肿瘤医学》2017年第8期1196-1203,共8页Journal of Modern Oncology

摘　　要：目的:研究FGFR1(成纤维生长因子受体1)抑制剂Ponatinib对乳腺癌细胞增殖和凋亡的影响以及机制。方法:依据前期Western Blot结果筛选出实验组和对照组,用不同浓度Ponatinib处理人乳腺癌细胞株实验组和对照组。CCK-8法检测乳腺癌细胞增殖抑制率;流式细胞术检测乳腺癌细胞凋亡的影响;Western Blot法检测Poantinib对FGFR1及其下游通路及凋亡相关蛋白的影响。结果:依据前期Western Blot结果,选取不表达FGFR1的MCF-7为对照组细胞,高表达FGFR1的MDA-MB-231、BT-549为实验组细胞。CCK-8结果及Annexin V-APC/7-AAD法结合流式细胞仪检测结果显示:Ponatinib呈浓度依赖性地诱导MDA-MB-231、BT-549细胞的增殖抑制和凋亡,对MCF-7细胞也有一定的增殖抑制和诱导凋亡作用,实验组细胞的增殖抑制率及凋亡率较对照组细胞高。Western Blot结果显示:MDA-MB-231、BT-549细胞中,pFGFR1、p-Stat5、p-PLCγ1蛋白表达量均明显下调;Bcl-2、Mcl-1表达量下调,Bak表达量上调(BT-549中Bak、Bax表达量均上调)。MCF-7细胞中,p-Stat5、p-PLCγ1蛋白表达量下调。结论:Ponatinib抑制MDA-MB-231、BT-549细胞增殖可能与下调p-FGFR1,进而下调p-Stat5、p-PLCγ1有关,促进细胞凋亡的机制可能与下调Bcl-2、Mcl-1,上调Bak有关。Objective： To investigate the FGFR1 inhibitor Ponatinib＇s effect on proliferation,apoptosis,preliminary explore its mechanism at protein level of breast cancer cells.Methods： Divided the experimental group and control group on the basis of expression levels of FGFR1.CCK-8 assay was used to detect the influence of Ponatinib on cell proliferation rate and AnnexinⅤ-APC /7-AAD assay was used to detect on celluar apoptosis.Western Blot assay was used to detect the influence of Ponatinib on the proteins related to FGFR1 signal transduction pathways and apoptosis.Results： FGFR1 was highly expressed in MDA-MB-231,BT-549 cells,the expression level of MCF-7 was few.So MCF-7,MDA-MB-231,BT549 cells were screened for the next experiments.The result of CCK8 assay and Annexin V-APC /7-AAD： Ponatinib could inhibit the proliferation and increase celluar apoptosis rates of MCF-7,MDA-MB-231 and BT-549 as the increasement of Ponatinib concentration.The apoptosis rates of experimental group were higher than control group,and the inhibition of MCF-7 proliferation was lower than MDA-MB-231 and BT-549.The result of Western Blot assay： Western-Blot results showed that the expression of p-FGFR1,p-Stat5 and p-PLCγ1 were declining in MDA-MB-231 and BT-549 cells.In the two cells,the protein expression levels of Bcl-2 and Mcl-1 were declining,Bak was increasing（Bak and Bax increased in BT-549）.The protein expression levels of p-Stat5,p-PLCγ1 declined in MCF-7 cells.Conclusion： The mechanism of inhibiting proliferation may relate to the decreased expression levels of p-Stat5,p-PLCγ1,which may induced by down-regulation of p-FGFR1.The mechanism of promoting apoptosis may relate to down-regulation of Bcl-2,Mcl-1,up-regulation of Bak.

关 键 词：FGFR1 PONATINIB 乳腺癌细胞 增殖 凋亡 

分 类 号：R737.9[医药卫生—肿瘤]