蟾蜍他灵对人结直肠癌细胞HCT116增殖、迁移、侵袭和上皮间质转化的影响  

Effect of bufotalin on the proliferation,migration,invasion,and epithelial-mesenchymal transition of human colorectal cancer HCT116 cells

作　　者：王艳 王莎莎[2] 朱春阳 董晨 王瑞 邱文生[2] WANG Yan;WANG Shasha;ZHU Chunyang;DONG Chen;WANG Rui;QIU Wensheng(Faculty of Medicine,Qingdao University,Qingdao 266071,China)

机构地区：[1]青岛大学医学部,山东青岛266071 [2]青岛大学附属医院肿瘤内科

出　　处：《精准医学杂志》2024年第2期120-124,共5页Journal of Precision Medicine

基　　金：北京市希思科临床肿瘤学研究基金(Y-HR-2018-185);青岛市中医药科技项目(2021-zyym29)。

摘　　要：目的探讨不同浓度蟾蜍他灵(BT)对人结直肠癌细胞HCT116增殖、迁移、侵袭和上皮间质转化的影响。方法采用CCK-8实验检测不同浓度(0、10、20、40、80、160、320 nmol/L)的BT处理24和48 h后的细胞活力,并计算半抑制浓度(IC 50);平板克隆实验验证0、12.5、25.0 nmol/L BT(设为A、B、C组)处理14 d后对HCT116细胞集落形成能力的影响;使用划痕和Transwell实验验证0、25、50 nmol/L BT(设为A、C、D组)处理24 h后对HCT116细胞迁移和侵袭能力的影响;应用Western blot实验检测A、C、D组处理24 h后HCT116细胞中E-cadherin和N-cadherin蛋白表达水平。结果CCK-8实验结果显示,BT处理HCT116细胞24或48 h后,随着BT的浓度增加,其抑制HCT116细胞增殖的作用显著增强(F=2106.00、3725.00,P<0.05),处理24和48 h的IC 50分别为49.59、24.10 nmol/L;平板克隆实验结果显示,B、C组细胞的集落数量显著少于A组(F=159.30,t=12.40、17.32,P<0.05);划痕和Transwell实验结果显示,C、D组细胞迁移率和侵袭细胞数量显著低于A组(F=120.30、296.80,t=12.71~21.27,P<0.05);Western blot实验结果显示,BT显著上调HCT116细胞内E-cadherin蛋白表达(F=2736.00,P<0.05),其中C、D组显著高于A组(t=50.27、72.13,P<0.05);而BT显著下调N-cadherin蛋白表达(F=626.80,P<0.05),其中C、D组显著低于A组(t=26.54、33.57,P<0.05)。结论BT可明显抑制人结直肠癌细胞HCT116的增殖、迁移、侵袭和上皮间质转化过程,有望成为结直肠癌治疗的潜在候选药物。Objective To investigate the effect of different concentrations of bufotalin(BT)on the proliferation,migration,invasion,and epithelial-mesenchymal transition of human colorectal cancer HCT116 cells.Methods CCK-8 assay was used to measure cell viability after 24 and 48 h of BT treatment at different concentrations(0,10,20,40,80,160,and 320 nmol/L),and the half-maximal inhibitory concentration(IC 50)was calculated.The plate colony formation assay was used to verify the effect of BT treatment at the concentrations of 0,12.5,and 25.0 nmol/L for 14 days on the colony formation ability of HCT116 cells(established as groups A,B,and C).The wound healing assay and the Transwell assay were used to verify the effect of BT treatment at the concentrations of 0,25,and 50 nmol/L for 24 h on the migration and invasion abilities of HCT116 cells(established as groups A,C,and D).Western blot was used to measure the protein expression levels of E-cadherin and N-cadherin in HCT116 cells after 24 h of treatment in groups A,C and D.Results CCK-8 assay showed that after HCT116 cells were treated by BT for 24 or 48 h,the inhibitory effect of BT on the proliferation of HCT116 cells increased significantly with the increase in the concentration of BT(F=2106.00,3725.00,P<0.05),with an IC 50 value of 49.59 nmol/L for 24-hour treatment and 24.10 nmol/L for 48-hour treatment.The plate colony formation assay showed that groups B and C had a significantly lower number of colonies of cells than group A(F=159.30,t=12.40,17.32,P<0.05).The wound healing assay and the Transwell assay showed that compared with group A,groups C and D had significantly lower cell migration rate and number of invading cells(F=120.30,296.80,t=12.71-21.27,P<0.05).Western blot showed that BT significantly upregulated the protein expression level of E-cadherin in HCT116 cells(F=2736.00,P<0.05),and groups C and D had a significantly higher expression level than group A(t=50.27,72.13,P<0.05);BT significantly downregulated the protein expression level of N-cadherin(F=626.80,P<0

关 键 词：结直肠肿瘤 HCT116细胞 蟾酥甾类 细胞增殖 细胞运动 肿瘤浸润 上皮-间质转化 

分 类 号：R735.34[医药卫生—肿瘤]