通关藤联合XELOX方案促进人结直肠癌HCT116细胞双硫死亡的作用  

Effect of Marsdenia tenacissima combined with XELOX solution on disulfide apoptosis in human colorectal cancer HCT116 cells

作　　者：韦伟 蔡曌颖 钱亚云 Wei Wei;Cai Zhaoying;Qian Yayun(Department of Gastrointestinal Surgery,Jiangdu People's Hospital Affiliated to Yangzhou University,Yangzhou 225200,China;Department of Integrated Traditional Chinese and Western Medicine,Medical College of Yangzhou University,Yangzhou 225009,China)

机构地区：[1]扬州大学附属江都人民医院普外科,扬州225200 [2]扬州大学医学院中西医结合学系,扬州225009

出　　处：《国际肿瘤学杂志》2024年第9期545-555,共11页Journal of International Oncology

基　　金：江苏省中医药科技发展计划(MS2021081);扬州市科技计划项目(YZ2022089、YZ2023094);扬州市卫生健康委医学科研重点项目(2023-1-04)。

摘　　要：目的探讨通关藤联合XELOX方案对人结直肠癌HCT116细胞双硫死亡的作用及相关机制。方法体外培养结直肠癌HCT116细胞,分别用0、0.3、0.6、1.2、2.4、4.8μg/ml浓度的卡培他滨,0、10、20、40、80、160μg/ml浓度的奥沙利铂,0、15、30、60、120、240 mg/ml浓度的通关藤,0、4、8、16、32、64μg/ml浓度的葡萄糖抑制剂BAY-876处理HCT116细胞;以及经25μg/ml BAY-876预处理后再经以上各药物、各浓度处理HCT116细胞。将HCT116细胞分为阴性对照组(不做任何处理)、卡培他滨组(4.0μg/ml)、奥沙利铂组(90μg/ml)、通关藤组(140 mg/ml)、XELOX方案组(4.0μg/ml卡培他滨+90μg/ml奥沙利铂)、通关藤联合XELOX方案组(140 mg/ml通关藤+4.0μg/ml卡培他滨+90μg/ml奥沙利铂)。BAY-876处理组为以上各组别中,每组加入葡萄糖抑制剂BAY-87625μg/ml。采用MTT法检测细胞增殖情况,AnnexinⅤ-FITC/PI双染法检测细胞凋亡情况,邻甲苯胺法检测葡萄糖浓度,NADPH比色法检测NADPH水平,胱氨酸摄取荧光法检测胱氨酸荧光强度,半胱氨酸比色法检测半胱氨酸含量。结果经0、0.3、0.6、1.2、2.4、4.8μg/ml浓度卡培他滨,0、10、20、40、80、160μg/ml浓度奥沙利铂,0、4、8、16、32、64μg/ml浓度BAY-876,以及经25μg/ml葡萄糖抑制剂BAY-876预处理后再经上述各药物、各浓度处理HCT116细胞后,细胞存活率差异均有统计学意义(F=644.60,P<0.001;F=417.30,P<0.001;F=1028.00,P<0.001;F=1066.00,P<0.001;F=847.70,P<0.001),且随着各药物浓度增加,HCT116细胞活性均逐渐下降(均P<0.05);经0、15、30、60、120、240 mg/ml浓度通关藤,以及经25μg/ml葡萄糖抑制剂BAY-876预处理后再经上述浓度药物处理HCT116细胞后,细胞存活率差异均具有统计学意义(F=107.50,P<0.001;F=619.70,P<0.001),且随着药物浓度增加,HCT116细胞活性呈现先上升后下降的趋势(均P<0.05)。AnnexinⅤ-FITC/PI双染法结果显示,阴性对照组绿色、红色、黄色荧光强度均较弱,�Objective To investigate the effect and related mechanism of Marsdenia tenacissima combined with XELOX solution on disulfide apoptosis in human colorectal cancer HCT116 cells.Methods The human colorectal cancer HCT116 cells were cultured in vitro and treated with different concentrations of capecitabine(0,0.3,0.6,1.2,2.4,4.8μg/ml),oxaliplatin(0,10,20,40,80,160μg/ml),Marsdenia tenacissima(0,15,30,60,120,240 mg/ml),and the glucose inhibitor BAY-876(0,4,8,16,32,64μg/ml),respectively.Furthermore,the HCT116 cells were pre-treated with 25μg/ml of BAY-876,followed by exposure to the specified concentrations of each drug group.HCT116 cells were divided into the following groups:negative control group(no treatment),capecitabine group(4.0μg/ml),oxaliplatin group(90μg/ml),Marsdenia tenacissima group(140 mg/ml),XELOX solution group(4.0μg/ml capecitabine+90μg/ml oxaliplatin),and Marsdenia tenacissima combined with XELOX solution group(140 mg/ml Marsdenia tenacissima+4.0μg/ml capecitabine+90μg/ml oxaliplatin).BAY-876 treatment groups refer to the groups which the glucose inhibitor BAY-87625μg/ml was added to each of the above groups.The cell proliferation was assessed using the MTT assay.Apoptosis was determined through AnnexinⅤ-FITC/PI double staining.The concentration of glucose was quantified using the o-toluidine method.NADPH levels were measured by colorimetry.Cystine uptake fluorescence assay was utilized to quantify the fluorescence intensity of cystine,and cysteine content was determined using cysteine colorimetry.Results After 0,0.3,0.6,1.2,2.4,4.8μg/ml concentration of capecitabine,0,10,20,40,80,160μg/ml concentration of oxaliplatin,0,4,8,16,32,64μg/ml concentration of BAY-876,and after pre-treatment with 25μg/ml glucose inhibitor BAY-876,HCT116 cells were treated with the above drugs and concentrations,there were statistically significant differences in cell survival rate(F=644.60,P<0.001;F=417.30,P<0.001;F=1028.00,P<0.001;F=1066.00,P<0.001;F=847.70,P<0.001),and with the increase of each drug concen

关 键 词：结直肠肿瘤 卡培他滨 奥沙利铂 通关藤 双硫死亡 

分 类 号：R285[医药卫生—中药学]