华蟾素对乳腺癌MDA-MB-231、MCF-7细胞增殖及p-PTEN表达的影响

Effects of cinobufacin on proliferation and expression of p-PTEN in breast cancer MDA-MB-231 and MCF-7 cells

目的观察华蟾素对人乳腺癌MCF-7、MDA-MB-231细胞人第10号染色体缺失的磷酸酶和张力蛋白的同源基因(phosphatase and tensin homolog deleted on chromosome ten,PTEN)蛋白表达的影响。方法将MCF-7、MDA-MB-231细胞按随机数字表法分为对照组、高剂量组、低剂量组。低、高剂量组分别加入0.5、5.0 mg/ml华蟾素注射液l00μl/孔进行干预;对照组加入等体积的RPMI-1640培养液。干预后6、12、24、48、72 h,采用细胞计数试剂盒-8法检测细胞增殖率,干预后72 h,采用Western blot法检测蛋白激酶B/哺乳动物雷帕霉素靶蛋白(protein kinase B/mammalian target of rapamycin,AKT/mTOR)通路蛋白及p-PTEN蛋白表达。结果与对照组比较,干预后24、48、72 h,高剂量组MCF-7、MDA-MB-231细胞增殖率降低(P<0.05);低、高剂量组MCF-7、MDA-MB-231细胞p-AKT[MCF-7:(0.357±0.064)、(0.215±0.056)比(0.924±0.085);MDA-MB-231:(0.310±0.022)、(0.194±0.019)比(0.811±0.089)]、p-mTOR[MCF-7:(0.476±0.039)、(0.217±0.038)比(0.838±0.058);MDA-MB-231:(0.300±0.031)、(0.223±0.025)比(0.896±0.096)]、p-S6[MCF-7:(0.551±0.068)、(0.428±0.041)比(1.254±0.264);MDA-MB-231:(0.281±0.014)、(0.197±0.012)比(0.748±0.022)]、p-PTEN[MCF-7:(0.487±0.170)、(0.184±0.135)比(1.003±0.284);MDA-MB-231:(0.261±0.184)、(0.170±0.105)比(1.014±0.206)]表达降低(P<0.05)。结论华蟾素可抑制MCF-7、MDA-MB-231细胞增殖,推测可能与其下调AKT/mTOR信号通路上游蛋白p-PTEN蛋白表达有关。

Objective To observe the effect of cinobufacin on the expression of phosphorylated PTEN protein in human breast cancer MCF-7 and MDA-MB-231 cells.Methods The MCF-7 and MDA-MB-231 cells were divided into control group,high dose group and low dose group according to random number table method.Low and high dose groups were added with 0.5,5.0 mg/ml cinobufacin,100μl/well for intervention,respectively,while the control group was added with equal volume of RPMI-1640 medium.Cell Counting Kit-8 was used to detect cell proliferation assay at 24,48,and 72 h following intervention.Immunoblotting was used to detect the expression of AKT/mTOR pathway protein expression and p-PTEN.Results At 24,48,and 72 h after intervention,the proliferation of MCF-7 and MDA-MB-231 cells in high dose group were significantly decreased(P<0.05).Compared with the control group,the expression of p-AKT(MCF-7:0.357±0.064,0.215±0.056 vs.0.924±0.085;MDA-MB-231:0.310±0.022,0.194±0.019 vs.0.811±0.089),p-mTOR(MCF-7:0.476±0.039,0.217±0.038 vs.0.838±0.058;MDA-MB-231:0.300±0.031,0.223±0.025 vs.0.896±0.096),p-S6(MCF-7:0.551±0.068,0.428±0.041 vs.1.254±0.264;MDA-MB-231:0.281±0.014,0.197±0.012 vs.0.748±0.022),p-PTEN(MCF-7:0.487±0.170,0.184±0.135 vs.1.003±0.284,P<0.05;MDA-MB-231:0.261±0.184,0.170±0.105 vs.1.014±0.206)in the low and high dose group significantly decreased(P<0.05).Conclusions Cinobufacin inhibited the proliferation of MCF-7 and MDA-MB-231 cells,which may be mediated by down-regulation of the expression of the upstream of the AKT/mTOR signaling pathway p-PTEN.