二甲双胍拮抗STAT3信号通路在人乳腺癌顺铂耐药细胞凋亡中的作用  

Effect of metformin antagonizing STAT3 signaling pathway in apoptosis of human breast cancer cisplatin-resistant cells

作　　者：赵亮[1] 吕宏程 王海龙 ZHAO Liang;LYU Hong-Cheng;WANG Hai-Long(Department of Surgery,Affiliated Hospital of Tianjin Academy of Traditional Chinese Medicine,Tianjin,300120,China)

机构地区：[1]天津市中医药研究院附属医院外科,天津300120 [2]天津市中医药研究院附属医院肿瘤科,天津300120

出　　处：《中国妇幼保健》2022年第8期1490-1495,共6页Maternal and Child Health Care of China

基　　金：天津市科学技术局项目(津20201103)。

摘　　要：目的 分析二甲双胍拮抗信号传导与转录活化因子3(STAT3)信号通路在人乳腺癌顺铂耐药细胞凋亡中的作用。方法 MCF-7细胞株为A组,MCF-7/DDP细胞株为B组,MCF-7/DDP细胞株+DDP为C组,MCF-7/DDP细胞株+二甲双胍为D组,MCF-7/DDP细胞株+STAT3抑制剂STA-21为E组。A组和B组采用正常培养基培养,C组MCF-7/DDP细胞株加入1μg/ml DDP培养,D组MCF-7/DDP细胞株加入5 mmol/L二甲双胍培养,E组加入10μmoL/L的STAT3抑制剂STA-21培养。对MCF-7细胞株和MCF-7/DDP细胞株采用CCK-8法检测DDP的敏感性;采用平板克隆试验检测各组细胞增殖数目;采用流式细胞仪检测各组细胞凋亡率;采用免疫荧光法检测各组受体相互作用蛋白(RIP)1和RIP 3蛋白表达水平;采用免疫印迹法检测各组细胞STAT3、p-STAT3、Caspase及p-Caspase蛋白表达水平。结果 浓度分别为0.01μg/ml、0.1μg/ml、1μg/ml、10μg/ml及100μg/ml的DDP对MCF-7细胞IC50值均高于MCF-7/DDP细胞,差异均有统计学意义(均P<0.05)。A组、B组、C组、D组及E组MCF-7/DDP细胞增殖数目分别为(152.51±10.63)个、(182.29±15.71)个、(124.53±8.62)个、(86.03±5.40)个及(85.71±6.12)个,差异有统计学意义(F=107.900,P<0.001)。A组、B组、C组、D组及E组MCF-7/DDP细胞凋亡率分别为(6.81±0.54)%、(6.62±0.50)%、(17.78±2.22)%、(27.59±4.62)%及(28.45±3.97)%,差异有统计学意义(F=80.050,P<0.001)。A组和B组细胞核完整,呈均匀蓝色;C组细胞核浓染,出现了细胞核碎裂;D组和E组细胞核浓染加深,核碎裂现象显著。与B组比较,C组RIP 1和RIP 3蛋白表达水平显著升高(均P<0.05);与C组比较,D组RIP 1和RIP 3蛋白表达水平显著升高(均P<0.05)。与A组比较,B组细胞STAT3、p-STAT3蛋白表达水平显著升高(均P<0.05),Caspase3、p-Caspase3蛋白表达水平显著降低(均P<0.05);与B组比较,C组细胞STAT3、p-STAT3蛋白表达水平显著降低(均P<0.05),Caspase3、p-Caspase3蛋白表达水平显著升高(均P<0.05);与C组比较,D组细胞STAObjective To analyze the effect of metformin antagonizing signal transducer and activator of transcription 3(STAT3) signaling pathway in apoptosis of human breast cancer cisplatin-resistant cells. Methods MCF-7 cell line was designed as A group, MCF-7/DDP cell line was designed as B group, MCF-7/DDP cell line+DDP was designed as C group, MCF-7/DDP cell line+metformin was designed as D group, MCF-7/DDP cell line+STAT3 inhibitor STA-21 was designed as E group. The cells in A group and B group were cultured using normal medium, 1 μg/ml DDP was added into MCF-7/DDP cells in C group, 5 mmol/L metformin was added into MCF-7/DDP cells in D group, 10 μmol/L STAT3 inhibitor STA-21 was added into the cells in E group. The sensitivities of DDP in MCF-7 cell line and MCF-7/DDP cell line were detected by CCK-8 assay;plate cloning experiment was used to detect the numbers of cells in the five groups;flow cytometry was used to detect cell apoptotic rate;immumofluorescence method was used to detect receptor interacting protein(RIP) and RIP 3;western blot was used to detect STAT3, p-STAT3, Caspase, and p-Caspase proteins in the five groups. Results IC50 values of 0.01 μg/ml, 0.1 μg/ml, 1 μg/ml, 10 μg/ml, and 100 μg/ml DDP in MCF-7 cells were statistically significantly higher than those in MCF-7/DDP cells(P<0.05). The proliferation numbers of MCF-7/DDP cells in A group, B group, C group, D group, and E group were(152.5±10.6),(182.3±15.7),(124.5±8.6),(86.0±5.4), and(85.7±6.1), respectively, there was statistically significant difference(F=107.900, P<0.001). The apoptotic rates of MCF-7/DDP cells in A group, B group, C group, D group, and E group were(6.81±0.54)%,(6.62±0.50)%,(17.78±2.22)%,(27.59±4.62)%, and(28.45±3.97)%, respectively, there was statistically significant difference(F=80.050, P<0.001). In A group and B group, the cell nucleus was intact, showing uniform blue;in C group, nuclear concentration and fragmentation were observed;in D group and E group, nuclear concentration and fragmentation were more sig

关 键 词：乳腺癌 顺铂耐药 二甲双胍 信号传导与转录活化因子3 

分 类 号：R737.9[医药卫生—肿瘤]