BGJ398与顺铂单独或联合作用对宫颈癌He La细胞增殖、凋亡、侵袭的影响及其机制研究

Effects of BGJ398 and cisplatin alone or in combination on proliferation, apoptosis and invasion of cervical cancer HeLa cells and its mechanism

目的探讨成纤维细胞生长因子受体抑制剂BGJ398与顺铂单独或联合作用对宫颈癌HeLa细胞增殖、凋亡、侵袭的影响及可能的作用机制。方法培养宫颈癌HeLa细胞,分别用不同浓度的BGJ398(0、10、20、30、40μmol/L)和顺铂(0、10、20、30、40、50μmol/L)处理24、48、72h。采用CCK-8试剂盒检测细胞增殖抑制率,根据结果选择12.5μmol/L作为实验药物浓度,将宫颈癌HeLa细胞分为对照组(无BGJ398和顺铂的培养基)、BGJ398组(含有12.5μmol/L BGJ398的培养基)、顺铂组(含有12.5μmol/L顺铂的培养基)、联合组(同时含有12.5μmol/L BGJ398和12.5μmol/L顺铂的培养基),进行后续实验,再次用CCK-8试剂盒检测各组细胞的增殖抑制率,采用流式细胞术检测细胞凋亡率,采用Transwell小室检测细胞侵袭能力,采用Westernblotting法检测细胞增殖、凋亡、侵袭相关蛋白的表达以及PI3K/AKT和MEK/ERK信号通路蛋白的表达。结果 BGJ398组、顺铂组、联合组的细胞增殖抑制率高于对照组,联合组的细胞增殖抑制率高于BGJ398组、顺铂组,差异均有显著性(P<0.05)。BGJ398组、顺铂组、联合组的细胞凋亡率高于对照组,联合组的细胞凋亡率高于BGJ398组、顺铂组,差异均有显著性(P<0.01)。BGJ398组、顺铂组、联合组穿膜细胞个数低于对照组,联合组的穿膜细胞数低于BGJ398组、顺铂组,差异均有显著性(P<0.01)。BGJ398组、顺铂组、联合组细胞的PCNA、MMP-2表达量低于对照组,Bax、E-cadeherin表达量高于对照组,差异均有显著性(P<0.01)。联合组PCNA、MMP-2表达量低于BGJ398组、顺铂组,Bax、E-cadeherin表达量高于BGJ398组、顺铂组,差异均有显著性(P<0.01)。BGJ398组p-ERK1/2、p-AKT表达量低于对照组(P<0.01),ERK1/2、AKT表达量与对照组比较差异无显著性(P>0.05)。结论 BGJ398与顺铂单独或联合作用均能抑制宫颈癌HeLa细胞的增殖和侵袭能力,促进细胞凋亡,其机制可能与抑制p-AKT、p-ERK1/2、PCNA、MMP-2蛋白表达及促进Bax、E-cadeherin蛋白表达有关。

Objective To explore the effect and mechanism of fi broblast growth factor receptor inhibitor BGJ398 alone and in combination with cisplatin on the proliferation, apoptosis and invasion of cervical cancer HeLa cells. Method Culture cervical cancer HeLa cells, they were treated with different concentrations of BGJ398(0,10,20,30,40) μmol/L,cisplatin(0,10,20.,30,40,50) μmol/L for 24 h, 48 h, 72 h. Then set the control group, BGJ398 group(12.5μmol/L BGJ398), cisplatin group(12.5μmol/L cisplatin), combined group(12.5μmol/L BGJ398 combined with 12.5μmol/L cisplatin). The cellular proliferation inhibition rate of each group are detected by CCK-8 method, the apoptosis rate are detected by fl ow cytometry, the invasion ability of cells are detected by Transwell chamber technology, and the expression of cell proliferation-related protein PCNA, apoptosis-related protein Bax, invasion-related protein MMP-2,E-cadherin and PI3K-AKT signal pathway-related protein p-AKT, AKT and MEK/ERK signal pathway-related protein p-ERK,ERK were detected by Western blot. Result The cellular proliferation inhibition rate of BGJ398 group,cisplatin group and combined group were higher than control group(P<0.05). The cellular proliferation inhibition rate of combined group was higher than BGJ398 group and cisplatin group(P<0.01). The apoptosi s rate of BGJ398 group,cisplatin group and combined group were higher than control group(P<0.01), among the three experimental groups the combined group was higher than BGJ398 group and cisplatin group(P<0.01).The number of trans membrane cell of BGJ398 group,cisplatin group and combined group were lower than control group(P<0.01), of which the combined group was lower than BGJ398 group and cisplatin group(P<0.01). The expression of PCNA and MMP-2 in BGJ398 group,cisplatin group and combined group were lower than control group(P<0.01), and the expression of Bax and E-cadeherin in the three experimental groups were higher than control group(P<0.01). The expression of PCNA and MMP-2 in combined group were lower than BGJ398 group,cisplatin group(P<0.01), and the expression of Bax and E-cadeherin in it were higher than the other two groups(P<0.01). The expression of p-ERK1/2 and p-AKT in BGJ398 group were lower than control group(P<0.01). There was no signifi cant difference in the expression of ERK1/2 and AKT between Control group and BGJ398 groups(P>0.05). Conclusion BGJ398 synergize with cisplatin can inhibit the proliferation, invasion and apoptosis of Cervical Cancer HeLa cells. The mechanism may be related to the inhibition of p-AKT, p-ERK1/2, PCNA, MMP-2 and the promotion of Bax and E-cadeherin expression.