当归多糖对结直肠癌细胞顺铂耐药性的影响及作用机制  

Effect of Angelica sinensis Polysaccharides on Cisplatin Resistance in Colorectal Cancer Cells and Its Mechanism of Action

作　　者：杨柳[1,2] 彭飞 陆平 傅永吉[1] YANG Liu;PENG Fei;LU Ping;FU Yongji(Affiliated Ninth People’s Hospital,School of Medicine,Shanghai Jiaotong University,Shanghai 200011,China;Shanghai Key Laboratory of Gut Microecology and Associated Major Diseases Research,Shanghai 200011,China;Shanghai Jing’an District Central Hospital,Shanghai 200040,China)

机构地区：[1]上海交通大学医学院附属第九人民医院,上海200011 [2]上海市消化道微生态及相关重大疾病研究重点实验室,上海200011 [3]上海市静安区中心医院,上海200040

出　　处：《北华大学学报（自然科学版）》2024年第4期448-453,共6页Journal of Beihua University（Natural Science）

基　　金：上海市卫健委卫生行业临床研究基金项目(202040436)。

摘　　要：目的 探讨当归多糖对结直肠癌细胞顺铂耐药性的影响及作用机制。方法 将人结直肠癌细胞HT-15、顺铂耐药细胞株HT-15/DDP仅使用30μmol/L浓度的顺铂干预设为对照组,使用对应质量浓度1.0、1.5、2.0 mg/mL的当归多糖干预12 h后再加入30μmol/L浓度顺铂设为当归多糖组。通过脂质体细胞转染技术分别将miR-10b-5p inhibitor、TGFBR2 mimics转染至HT-15/DDP细胞中,再加入2.0 mg/mL当归多糖干预48 h,然后将其分为当归多糖(2.0 mg/mL)+miR-10b-5p inhibitor组、当归多糖(2.0 mg/mL)+TGFBR2 mimics组。MTT法检测细胞增殖能力;Transwell小室试验检测细胞迁移能力;流式细胞术检测细胞凋亡能力。RT-PCR检测miR-10b-5p、TGFBR2 mRNA表达水平;Western blot检测TGFBR2蛋白表达水平。结果 单独顺铂干预的对照组HT-15/DDP细胞增殖率、细胞迁移个数明显高于HT-15细胞(P<0.01);经不同质量浓度当归多糖预处理12 h后,可明显增强HT-15/DDP细胞对顺铂的敏感性,HT-15/DDP细胞增殖率、细胞迁移个数明显降低(P<0.01)。单独顺铂干预的对照组HT-15/DDP细胞凋亡率明显低于HT-15细胞(P<0.01);经不同质量浓度当归多糖预处理12 h后,明显增强了HT-15/DDP细胞对顺铂的敏感性,HT-15/DDP细胞凋亡率明显升高(P<0.01)。HT-15/DDP细胞miR-10b-5p、TGFBR2表达水平在各组间比较差异均具有统计学意义(P<0.01);不同剂量当归多糖组中HT-15/DDP细胞miR-10b-5p表达水平明显低于对照组,TGFBR2表达水平明显高于对照组(P<0.01),随着当归多糖剂量的增加miR-10b-5p表达水平逐渐降低,TGFBR2表达水平逐渐升高(P<0.01)。使用生物信息学数据库(TargetScan、miRanda)预测miR-10b-5p的靶基因,结果显示,TGFBR2是miR-10b-5p的候选靶基因。miR-10b-5p表达下调后,HT-15/DDP细胞中TGFBR2相对表达量明显高于对照组HT-15/DDP细胞中TGFBR2相对表达量(P<0.01)。当归多糖(2.0 mg/mL)+miR-10b-5p inhibitor组、当归多糖(2.0 mg/mL)+TGFBR2 mimics组的HT-15/DObjective To investigate the effect and mechanism of Angelica sinensis polysaccharides on cisplatin resistance in colorectal cancer cells.Method Divide human colorectal cancer cell line HT-15 and cisplatin resistant cell line HT-15/DDP into control group(using only 30μmol/L).Angelica sinensis polysaccharide group(1.0,1.5,2.0 mg/mL)was intervened with corresponding concentrations of Angelica sinensis polysaccharides for 12 hours,and adding 30μmol/L concentration cisplatin.MiR-10b-5p inhibitor and TGFBR2 mimics were transfected into HT-15/DDP cells by using liposome cell transfection technology.2.0 mg/mL Angelica sinensis polysaccharides were added for 48 hours of intervention,and were divided into two groups:Angelica sinensis polysaccharides(2.0 mg/mL)+miR-10b-5p inhibitor group and Angelica sinensis polysaccharides(2.0 m g/mL)+TGFBR2 mimics group.MTT assay was used to detect cell proliferation ability;Transwell chamber experiment to detect cell migration ability;Flow cytometry was used to detect the ability of apoptosis.RT-PCR was used to detect the expression levels of miR-10b-5p and TGFBR2 mRNA.Western blot was used to detect the expression level of TGFBR2 protein.Results The proliferation rate and cell migration number of HT-15/DDP cells in the control group treated with cisplatin alone were significantly higher than those in HT-15 cells(P<0.01);After 12 hours of pretreatment with different concentrations of Angelica sinensis polysaccharides,the sensitivity of HT-15/DDP cells to cisplatin was significantly enhanced,and the proliferation rate and migration number of HT-15/DDP cells were significantly reduced(P<0.01).The apoptosis rate of HT-15/DDP cells in the control group treated with cisplatin alone was significantly lower than that of HT-15 cells(P<0.01);After 12 hours of pretreatment with different concentrations of Angelica sinensis polysaccharides,the sensitivity of HT-15/DDP cells to cisplatin was significantly enhanced,and the apoptosis rate of HT-15/DDP cells was significantly increased(P<0.01).The

关 键 词：当归多糖 结直肠癌 HT-15细胞 HT-15/DDP细胞 顺铂耐药 miR-10b-5p TGFBR2 

分 类 号：R735.34[医药卫生—肿瘤]