小檗碱对结肠癌细胞增殖和侵袭的影响及机制  

Effect and mechanism of berberine on proliferation and invasion of colon cancer cells

作　　者：张舒 孟珊珊 张云鹤 蔡静 ZHANG Shu;MENG Shanshan;ZHANG Yunhe;CAI Jing(Department of Pathology,the First Affiliated Hospital of Henan University,Kaifeng 475001,Henan Province,China)

机构地区：[1]河南大学第一附属医院病理科,河南开封475001

出　　处：《新乡医学院学报》2024年第11期1001-1007,共7页Journal of Xinxiang Medical University

基　　金：河南省医学科技攻关计划项目(编号:LHGJ20200548)。

摘　　要：目的探讨小檗碱对人结肠癌细胞增殖、侵袭的影响及作用机制。方法选择2021年1月至2022年12月于河南大学第一附属医院行结肠癌根治术96例患者的石蜡包埋肿瘤组织和肠管切端组织(对照组织)为研究对象,应用免疫组织化学法检测肿瘤组织及对照组织中乏氧诱导因子-1α(HIF-1α)的表达,并分析HIF-1α与肿瘤临床病理特征的关系。将人结肠癌细胞株HCT116分别于常氧和乏氧环境下培养,待贴壁后分为常氧组、乏氧组、常氧联合小檗碱组、乏氧联合小檗碱组;常氧联合小檗碱组和乏氧联合小檗碱组分别加入25、50、75、100μmol·L^(-1)小檗碱,常氧组和乏氧组细胞不加任何药物;分别于培养24、48、72、96 h时,采用噻唑蓝比色法检测各组细胞增殖情况,根据结果确定最适合药物浓度为75μmol·L^(-1)。取常氧组、常氧联合75μmol·L^(-1)小檗碱组、乏氧组、乏氧联合75μmol·L^(-1)小檗碱组细胞,于培养24 h时,采用Transwell法检测各组细胞的侵袭能力;培养48 h时,采用Western blot法检测各组细胞中HIF-1α、血管内皮生长因子(VEGF)蛋白相对表达量。结果HIF-1α主要在结肠癌组织及对照组织细胞核表达。对照组织及结肠癌组织中HIF-1α阳性表达率分别为19.8%(19/96)、67.7%(65/96),结肠癌组织中HIF-1α阳性表达率显著高于对照组织(χ^(2)=11.298,P<0.05)。HIF-1α在结肠癌中的表达与肿瘤直径、有无淋巴结转移、TNM分期相关(P<0.05),与患者性别、年龄及肿瘤浸润深度、分化程度无关(P>0.05)。常氧组细胞培养24 h时呈对数生长,加入浓度25μmol·L^(-1)的小檗碱对细胞增殖无明显影响(P>0.05);与0μmol·L^(-1)小檗碱比较,加入浓度为50μmol·L^(-1)和75μmol·L^(-1)的小檗碱在培养48 h后细胞增殖活性显著降低(P<0.05),加入浓度100μmol·L^(-1)小檗碱从培养24 h后细胞增殖活性显著降低(P<0.05)。乏氧组细胞加入浓度25μmol·L^(-1)的小檗碱培养48 hObjective To explore the effect of berberine on the proliferation and invasion of human colon cancer cells and its mechanism.Methods The paraffin-embedded tumor tissues and intestinal incisal tissues(control tissues)of 96 patients who underwent radical resection of colon cancer at the First Affiliated Hospital of Henan University from January 2021 to December 2022 were selected as the research subjects.The expression of hypoxia-inducible factor-1α(HIF-1α)in tumor tissues and control tissues was detected by using the immunohistochemical method,and the relationship between HIF-1αand clinicopathological features was analyzed.The human colon cancer cell line HCT116 was cultivated in normoxic and hypoxic conditions,respectively;after adhesion,the cells were divided into normoxic group,hypoxic group,normoxic combined with berberine group,and hypoxic combined with berberine group.The cells in the normoxic combined with berberine group and the hypoxic combined with berberine group were treated with 25,50,75,and 100μmol·L^(-1) berberine,respectively;while the cells in the normoxic group and the hypoxic group were not treated with any drugs.At 24,48,72,and 96 hours of cultivation,the proliferation of cells in each group was detected by using the thiazolyl blue tetrazolium bromide method,respectively.Based on the above results,the most suitable drug concentration was determined as 75μmol·L^(-1).At 24 hours of cultivation,the invasion of cells in the normoxic group,normoxic combined with 75μmol·L^(-1) berberine group,hypoxic group,and hypoxic combined with 75μmol·L^(-1) berberine group was detected by using the Transwell method;at 48 hours of cultivation,the relative expression levels of HIF-1αand vascular endothelial growth factor(VEGF)proteins in the cells in each group were detected by using Western blot.Results HIF-1αwas mainly expressed in the nucleus of colon cancer tissues and control tissues.The positive expression rates of HIF-1αin control tissues and colon cancer tissues were 19.8%(19/96)and 67.7%(65/

关 键 词：小檗碱 结肠癌 乏氧诱导因子-1Α 细胞增殖 细胞侵袭 

分 类 号：R285[医药卫生—中药学]