[目的]探究成纤维细胞生长因子13(Fibroblast growth factor 13,FGF13)对乳腺癌细胞MCF7凋亡的调控作用.[方法]Western Blotting和免疫组织化学染色检测FGF13的表达;构建干扰和超表达FGF13的MCF7细胞株;TUNEL法和Western Blotting检测细胞凋亡;DCFH-DA荧光探针法检测细胞内活性氧(Reactive oxygen species,ROS).[结果]与正常乳腺细胞相比,FGF13在乳腺癌MCF7细胞中表达升高(0.16±0.01 vs 3.89±0.23,P＜0.001).干扰FGF13 后,MCF7 细胞凋亡率升高(2.20％±0.10％vs 10.79％±0.69％,P＜0.001),促凋亡蛋白 Bax(0.99±0.02 vs 2.56±0.02)、Caspase 3(1.00±0.01 vs 1.70±0.01)和 p53(0.98±0.01 vs 2.22±0.03)的表达上调(P＜0.001),而抗凋亡蛋白 Bcl-xl(0.99±0.01 vs 0.29±0.01)、Bcl-2(0.99±0.01 vs 0.27±0.01)的表达下调(P＜0.001).相反,超表达FGF13后,细胞凋亡率和促凋亡蛋白的表达均降低(P＜0.001),而抗凋亡蛋白的表达均升高(P＜0.001).而且,干扰FGF13的表达引起细胞内ROS水平升高(1 552.00±51.37 vs 2 714.00±55.19,P＜0.001),而超表达则导致ROS减少(2 102.83±65.29 vs 1 111.67±63.53,P＜0.001).[结论]FGF13 可能通过 ROS 介导抑制乳腺癌 MCF7 细胞凋亡.
FGF13 inhibits apoptosis through regulating intracellular ROS in MCF7 cells
[Objective]To investigate the effect of fibroblast growth factor 13(FGF13)on the regulation of apoptosis in breast cancer MCF7 cell line.[Method]Western Blotting and immunohistochemical staining were used to detect FGF13 expression.The MCF7 cell lines with stable interference and over-expression of FGF13 were constructed.TUNEL and western blot were applied to detect apoptosis.DCFH-DA fluorescence probe was used to measure cellular reactive oxygen species(ROS).[Re-sult]Compared with the health breast cells,FGF13 expression increased in MCF7 cells(0.16±0.01 vs 3.89±0.23,P＜0.001).Knockdown of FGF13 expression resulted in increase(P＜0.001)of apoptosis(2.20％±0.10％vs 10.79％±0.69％)and expression of pro-apoptotic proteins Bax(0.99±0.02 vs 2.56±0.02),Caspase 3(1.00±0.01 vs 1.70±0.01)and p53(0.98±0.01 vs 2.22±0.03),and decrease(P＜0.001)of anti-apoptotic preoteins Bel-xl(0.99±0.01 vs 0.29±0.01)and Bel-2(0.99±0.01 vs 0.27±0.01).On the contrary,over-expression of FGF13 led to decrease of apoptosis and expression of the pro-apoptotic proteins(P＜0.001),and increase of expression of the anti-apoptotic pretoe-ins(P＜0.001).Furthermore,knockdown of FGF13 expression yielded accumulation of ROS(1 552.00±51.37 vs 2 714.00±55.19,P＜0.001),while FGF13 overexpression led to decrease of ROS(2 102.83±65.29 vs 1 111.67±63.53,P＜0.001).[Conclusion]FGF13 perhaps inhibits apoptosis through regulating intracellular ROS in MCF7 cells.
breast cancerapoptosisFGF13reactive oxygen speciesanti-apoptotic proteinspro-apoptotic proteinsgene expressioncaspases
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