枸杞多糖联合奥沙利铂可逆转结肠癌干细胞的耐药  

Reversal effect of Lycium barbarum polysaccharide in combination with oxaliplatin on drug resistance of colon cancer stem cells

作　　者：艾芳芳 肖红燕[3] 汪芳[3] 朱永朝[4] 马丽君[5] Ai Fangfang;Xiao Hongyan;Wang Fang;Zhu Yongzhao;Ma Lijun(School of Clinical Medicine,Ningxia Medical University,Yinchuan 750004,Ningxia Hui Autonomous Region,China;Key Laboratory of Ningxia Ethnomedicine Modernization of Ministry of Education,Ningxia Medical University,Yinchuan 750004,Ningxia Hui Autonomous Region,China;People’s Hospital of Ningxia Hui Autonomous Region,Yinchuan 750004,Ningxia Hui Autonomous Region,China;Department of Surgery,General Hospital of Ningxia Medical University,Yinchuan 750004,Ningxia Hui Autonomous Region,China;Department of Human Anatomy,Histology and Embryology,School of Basic Medicine,Ningxia Medical University,Yinchuan 750004,Ningxia Hui Autonomous Region,China)

机构地区：[1]宁夏医科大学临床医学院,宁夏回族自治区银川市750004 [2]宁夏医科大学宁夏少数民族医药现代化教育部重点实验室,宁夏回族自治区银川市750004 [3]宁夏回族自治区人民医院,宁夏回族自治区银川市750004 [4]宁夏医科大学总医院外科学研究室,宁夏回族自治区银川市750004 [5]宁夏医科大学基础医学院人体解剖与组织胚胎学系,宁夏回族自治区银川市750004

出　　处：《中国组织工程研究》2024年第1期74-79,共6页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金项目(82060634),项目负责人:马丽君;宁夏重点研发计划一般项目(2018BEG03018),项目负责人:马丽君。

摘　　要：背景:结肠癌临床主要采用以氟尿嘧啶、伊立替康及奥沙利铂为基础的治疗方法,研究发现这些药物的转运与ABC转运蛋白G家族成员2(ATP-binding cassette transport protein of G2,ABCG2)等膜转运蛋白相关,然而当患者对这些化疗药物产生耐药后,ABCG2的高表达致使治疗效果明显下降,引起了结肠癌的耐药问题。临床急切需要新的药物及治疗方法来提高疗效,枸杞多糖具有广泛的生物学活性,将多糖作为抗肿瘤药物,可以克服化疗和放疗过程中杀死肿瘤细胞的同时对机体正常细胞的损伤。目的:通过体外实验探索枸杞多糖联合奥沙利铂对结肠癌干细胞耐药的逆转作用,进而探讨枸杞多糖逆转结肠癌耐药的可能分子机制。方法:选用结肠癌细胞株HCT116以及奥沙利铂耐药细胞株HCT116-OXR进行体外实验,采用CCK8检测细胞增殖情况确定枸杞多糖和奥沙利铂的最适干预浓度和干预时间,进一步分为HCT116对照组,HCT116-OXR空白处理组,枸杞多糖组(2.5 mg/mL枸杞多糖),奥沙利铂组(10μmol/L奥沙利铂),枸杞多糖+奥沙利铂组(2.5 mg/mL枸杞多糖+10μmol/L奥沙利铂),流式细胞仪检测细胞凋亡情况,免疫荧光和Western blot检测磷酸甘露糖异构酶(phosphomannose isomerase,PMI)和ABCG2的表达,Western blot检测PI3K,AKT,Bcl-2和Bax的蛋白表达水平。结果与结论:①HCT116-OXR相较HCT116对枸杞多糖更敏感(P<0.05);②与HCT116-OXR空白处理组比较,奥沙利铂联合枸杞多糖促进了HCT116-OXR细胞凋亡(P<0.05),Bcl-2蛋白表达明显下调(P<0.05),Bax蛋白表达明显上调(P<0.05),ABCG2、PMI、PI3K、AKT的蛋白表达明显下调(P<0.05);③结果表明枸杞多糖通过抑制PMI/PI3K/AKT通路逆转结肠癌耐药,为研究枸杞多糖增敏化疗作用的分子机制奠定了基础。BACKGROUND:Clinical treatment for colon cancer mainly includes fluorouracil,irinotecan and oxaliplatin-based therapy.Studies have shown that membrane transport proteins such as ATP-binding cassette transport protein of G2(ABCG2)mediate the transport of these drugs.However,when patients develop resistance to these chemotherapeutic drugs,the high expression of ABCG2 leads to a significant decrease in the therapeutic effect and raises the problem of drug resistance in colon cancer.New drugs and treatments are urgently needed to improve the efficacy.Lycium barbarum polysaccharide has a wide range of biological activities.It can be used as anti-tumor drug to overcome the damage to normal cells in the process of chemotherapy and radiotherapy in tumor patients.OBJECTIVE:To explore the reversal effect of Lycium barbarum polysaccharide in combination with oxaliplatin on colon cancer drug-resistant cells through in vitro experiments to investigate the possible molecular mechanism of Lycium barbarum polysaccharide reversal on colon cancer drug-resistant cells.METHODS:Colon cancer cell line HCT116 and oxaliplatin-resistant cell line HCT116-OXR were selected for in vitro experiments.The optimal intervention concentration and intervention time of Lycium barbarum polysaccharide and oxaliplatin were determined by CCK8 assay of cell proliferation.Samples were further divided into the HCT116 control group,HCT116-OXR blank treatment group,Lycium barbarum polysaccharide group(2.5 mg/mL Lycium barbarum polysaccharide),and oxaliplatin group(10μmol/L oxaliplatin),and Lycium barbarum polysaccharide+oxaliplatin group(2.5 mg/mL Lycium barbarum polysaccharide+10μmol/L oxaliplatin).Cell apoptosis was detected by flow cytometry.The protein expression levels of phosphomannose isomerase(PMI)and ABCG2 were detected by immunofluorescence and western blot assay.Phosphatidylinositol3-kinase(PI3K),protein kinase B(AKT),B-cell lymphoma 2(Bcl-2)and BCL2-Associated X(Bax)were detected by western blot assay.RESULTS AND CONCLUSION:(1)HCT116-OXR was mo

关 键 词：PMI ABCG2 PI3K AKT 枸杞多糖 结肠癌干细胞 耐药 凋亡 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学] R73