二硫化二砷与粉防己碱对三阴性乳腺癌细胞株HCC1937的协同作用  

Synergistic effect of arsenic disulfide and tetrandrine on triple negative breast cancer cell line HCC1937

作　　者：李建蓉 樊英怡 裴晓华[3] 王煊[1] 王春晖 LI Jianrong;FAN Yingyi;PEI Xiaohua;WANG Xuan;WANG Chunhui(Beijing University of Chinese Medicine,Beijing 100029,China;Bejing University of Chinese Medicine Third Affiliated Hospital,Beijing 100029,China;Xiamen Hospital of Dongzhimen Hospital Bejing University of Chinese Medicine,Xiamen 361001,China;Fangshan Hospital Beijing University of Chinese Medicine,Beijing 102400,China)

机构地区：[1]北京中医药大学,北京100029 [2]北京中医药大学第三附属医院,北京100029 [3]北京中医药大学东直门医院厦门医院,厦门361001 [4]北京中医药大学房山医院,北京102400

出　　处：《中华中医药杂志》2023年第10期4918-4923,共6页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金青年科学基金项目(No.82004367);北京市自然科学基金面上项目(No.7172127)。

摘　　要：目的:观察和验证二硫化二砷(As_(2)S_(2))和粉防己碱(Tet)对三阴性乳腺癌(TNBC)细胞株HCC1937的协同作用。方法:采用MTT方法检测As_(2)S_(2)、Tet对TNBC细胞株HCC1937的抑制率,利用中效原理(Chou-Talalay法)确定药物协同作用浓度;通过流式细胞技术观察分析两药对于HCC1937细胞凋亡的影响;通过ELISA方法检测药物对AQP9、ABCC2和P-gp含量的影响;通过q-PCR方法检测,协同作用浓度条件下,药物对细胞中Caspase-3、Survivin、Bid、Bax、Bcl-2、PARP表达的影响。结果:As_(2)S_(2)、Tet抑制HCC1937细胞增殖,诱导细胞凋亡,药物作用与浓度和作用时间呈正相关性(P<0.05);干预48 h,As_(2)S_(2)与Tet的合药浓度为3μg/mL+2μg/mL为相加作用(P<0.05);浓度为6μg/mL+4μg/mL为协同作用(P<0.05)。As_(2)S_(2)和Tet分别单用及合用干预48 h后,ELISA结果提示均可降低细胞AQP9、ABCC2和P-gp含量,协同作用浓度效果最好(P<0.05);q-PCR结果提示As_(2)S_(2)单用或两药合用可上调Caspase-3、Bax、Bid表达,下调Bcl-2、PARP表达,且协同作用浓度效果最好(P<0.05)。结论:As_(2)S_(2)和Tet可抑制HCC1937细胞增殖,诱导其凋亡,其作用机制可能与调控凋亡因子相关,且在协同作用浓度条件下合用两药效果明显优于单药作用;两药产生协同作用的机制可能与调控AQP9、ABCC2和P-gp有关。Objective:To observe and verify the synergistic effect of arsenic disulfide(As_(2)S_(2))and tetrandrine(Tet)on triple negative breast cancer(TNBC)cell line HCC1937.Methods:MTT was used to detect the inhibition rate of As_(2)S_(2)and Tet on TNBC cell line HCC937,and Chou Talalay method was used to determine the concentration of drug synergy.The effects of drugs on the apoptosis of HCC1937 cells were observed and analyzed by flow cytometry.ELISA was used to detect the effects of drugs on AQP9,ABCC2 and P-gp.And q-PCR was used to detect he expression of Caspase-3,Survivin,Bid,Bax,Bcl-2 and PARP in cells.Results:As_(2)S_(2)and Tet inhibited the proliferation and induced apoptosis of HCC1937 cells,the drug effect was positively correlated with concentration and time(P<0.05).After 48 h of intervention,the synergism concentration of arsenic disulfide and tetrandrine was 6μg/mL+4μg/mL(P<0.05).After 48 h of intervention,the expression of AQP9,ABCC2 and P-gp were were down-regulated,and combined was significantly better than the single(P<0.05),the synergistic concentration was superior;As_(2)S_(2)alone or in combination of the two drugs could up-regulate the expression of Caspase-3,Bax and Bid,and down-regulate the expression of Bcl-2 and PARP,and the synergistic concentration(P<0.05).Conclusion:As_(2)S_(2)and Tet(combined or single)can significantly inhibit HCC1937,and under the condition of synergistic,combined is superior to single.The mechanism for this effect relates to the apoptosis,As_(2)S_(2)and Tet can control apoptosis-related factors induce apoptosis of tumor cells.The mechanism of the synergistic effect of the two drugs is presumably based on affecting AQP9,ABCC2 and P-gp,which can affect the arsenic content and drug resistance in the cells.

关 键 词：二硫化二砷 粉防己碱 协同作用 三阴性乳腺癌 

分 类 号：R285[医药卫生—中药学]