卡培他滨节拍化疗联合依西美坦通过PI3K-AKT信号通路抑制乳腺癌细胞增殖的实验研究  

Trial study of capecitabine metronomic chemotherapy combined with exemestane inhibit proliferation of breast cancer cells by PI3K-AKT signaling pathway

作　　者：顾玉兰[1] 朱金莲 许晔琼 宛传丹 钱军[3] Gu Yulan;Zhu Jinlian;Xu Yeqiong;Wan Chuandan;Qian Jun(Department of Oncology,Changshu No.2 People's Hospital of Jiangsu Province,Changshu 215500,China;Central Laboratory,Changshu Medical Examination Institute,Changshu 215500,China;Department of Oncology,Jiangsu Province Hospital of Chinese Medicine,Affiliated Hospital of Nanjing University of Chinese Medicine,Nanjing 210029,China)

机构地区：[1]江苏省常熟市第二人民医院肿瘤科,215500 [2]江苏省常熟市医学检验所中心实验室,215500 [3]江苏省中医院南京中医药大学附属医院肿瘤科,南京210029

出　　处：《肿瘤研究与临床》2021年第6期401-407,共7页Cancer Research and Clinic

基　　金：北京医卫健康公益基金(B17527)。

摘　　要：目的:探讨卡培他滨节拍化疗联合依西美坦对乳腺癌MCF-7细胞增殖及PI3K-AKT信号通路的影响。方法:体外培养MCF-7细胞,分为对照组(加入不含药物的DMEM培养液)、30μmol/L依西美坦组和卡培他滨节拍化疗联合用药组[30μmol/L依西美坦联合不同浓度(50、33、17μmol/L)卡培他滨]。CCK-8法检测细胞增殖抑制率,计算药物半数抑制浓度(IC50)。流式细胞术检测各组细胞周期与细胞凋亡率的变化。蛋白质印迹法检测MCF-7细胞PI3K-AKT信号通路中相关蛋白的表达。结果:卡培他滨和依西美坦作用MCF-7细胞72 h的IC50分别为101.2μmol/L和60.6μmol/L。24、48 h 30μmol/L依西美坦联合50、33、17μmol/L卡培他滨组MCF-7细胞增殖抑制率均高于30μmol/L依西美坦组(均P<0.01);30μmol/L依西美坦组、30μmol/L依西美坦+50μmol/L卡培他滨组、30μmol/L依西美坦+33μmol/L卡培他滨组、30μmol/L依西美坦+17μmol/L卡培他滨组细胞凋亡率分别为(18.1±2.6)%、(34.6±3.0)%、(27.6±1.3)%、(23.1±1.6)%,差异有统计学意义(F=23.652,P<0.01)。与对照组相比,30μmol/L依西美坦组MCF-7细胞G 2期细胞比例上升[(16.7±2.6)%比(10.6±2.2)%],G 1期细胞比例下降[(53.3±4.0)%比(56.3±3.2)%];30μmol/L依西美坦+50μmol/L卡培他滨组MCF-7细胞S期细胞比例上升[(39.0±3.6)%比(33.1±2.0)%];30μmol/L依西美坦+33μmol/L卡培他滨组MCF-7细胞更多停滞于S期[(51.7±4.1)%],G 2期细胞几乎消失[(1.2±0.5)%];30μmol/L依西美坦+17μmol/L卡培他滨组MCF-7细胞G 2期细胞比例上升[(26.2±3.1)%]。蛋白质印迹法检测显示,低剂量节拍化疗促进依西美坦抑制PI3K表达,促进AKT蛋白473位丝氨酸磷酸化[p-AKT(473)表达增高],促进信号通路下游S6蛋白表达及提高磷酸化水平(p-S6表达升高),从而激活凋亡信号。结论:卡培他滨节拍化疗联合依西美坦可协同抑制乳腺癌MCF-7细胞增殖,通过影响PI3K-AKT信号通路激活细胞凋亡机制。Objective To investigate effects of capecitabine metronomic chemotherapy combined with exemestane on the proliferation of breast cancer MCF-7 cells and PI3-K/AKT signaling pathway.Methods MCF-7 cells cultured in vitro were divided into the control group(adding DMEM without drugs),30µmol/L exemestane group,capecitabine metronomic chemotherapy combined drugs group[30µmol/L exemestane combined with different concentrations(50,33,17µmol/L)of capecitabine].CCK-8 assay was used to detect the cell proliferation inhibition rate,the half-maximal inhibitory concentration(IC50)was calculated,and the changes of cell cycle and apoptosis rate of MCF-7 in different drug groups were assessed by using flow cytometry.The related-protein expression of PI3K-AKT signaling pathway of MCF-7 cells was detected by using Western blot.Results The IC50 of capecitabine and exemestane on MCF-7 cells for 72 h was 101.2μmol/L and 60.6µmol/L,respectively.The proliferation inhibition rate of MCF-7 cells in 30µmol/L exemestane for 24 h and 48 h combined with 50,33 and 17µmol/L capecitabine group was higher than that in 30µmol/L exemestane group(all P<0.01).The apoptosis rates were(18.1±2.6)%,(34.6±3.0)%,(27.6±1.3)%,(23.1±1.6)%,respectively in 30µmol/L exemestane group,30µmol/L exemestane+50µmol/L capecitabine group,30µmol/L exemestane+33µmol/L capecitabine group,30µmol/L exemestane+17µmol/L capecitabine group,and the difference was statistically significant(F=23.652,P<0.01).Compared with the control group,the proportion of MCF-7 cells in phase G2 of 30µmol/L exemestane group was increased[(16.7±2.6)%vs.(10.6±2.2)%],while that in phase G1 was decreased[(53.3±4.0)%vs.(56.3±3.2)%].The proportion of MCF-7 cells in phase S of 30µmol/L exemestane+50µmol/L capecitabine group was increased[(39.0±3.6)%vs.(33.1±2.0)%].MCF-7 cells of 30µmol/L of exemestane+33µmol/L capecitabine group were more blocked in phase S[(51.7±4.1)%],and cells in phase G2 were nearly disappeared[(1.2±0.5)%];the cell proportion MCF-7 cells in phase G2 o

关 键 词：乳腺肿瘤 抗肿瘤联合化疗方案 卡培他滨 依西美坦 

分 类 号：R737.9[医药卫生—肿瘤]