CD133抑制剂对人胆囊癌细胞的影响  

Effect of CD133 inhibitor on human gallbladder cancer cells

作　　者：郭跃 王少波 刘超[1] 王世行[1] 庞志刚[1] Guo Yue;Wang Shaobo;Liu Chao;Wang Shihang;Pang Zhigang(Department of General Surgery,the Second Affiliated Hospital of Zhengzhou University,Zhengzhou 450014,China)

机构地区：[1]郑州大学第二附属医院普外科,450014

出　　处：《中华实验外科杂志》2022年第7期1284-1287,共4页Chinese Journal of Experimental Surgery

摘　　要：目的观察Bufalin(蟾蜍灵)、AMD3100(普乐沙福)作为药物单独或联合作用于胆囊癌细胞株(GBC-SD)后,观察其对细胞增殖、迁移、凋亡以及CD133表达的影响,探讨其在胆囊癌中的作用。方法收集2017年1月至2021年12月郑州大学第二附属医院手术切除的胆囊癌标本20例,通过免疫组织化学法检测CD133在肿瘤以及癌旁组织的表达的差异。用不同浓度的Bufalin(80、160、320 nmol/L);AMD3100(10、20、40μmol/L)及联合用药(Bufalin 160 nmol/L+AMD310020μmol/L)干预人胆囊癌(GBC-SD)细胞,细胞计数试剂盒(CCK-8)法观察药物对肿瘤细胞增殖产生的影响,Transwell小室法检测肿瘤细胞迁移能力,膜联蛋白V(Annexin V)/碘化丙锭(PI)法检测细胞凋亡,流式细胞法检测细胞CD133分子表达。率的比较采用χ2检验,组间比较用单因素及双因素方差分析。结果免疫组织化学结果示CD133分子在人胆囊癌组织中的阳性表达率为85%,明显高于癌旁组织表达率[30%,χ2=10.23,P<0.01],表达定位于胞质和胞核,流式结果显示CD133在GBC-SD细胞中阳性表达。Bufalin(80、160、320 nmol/L)作用(24、48、72 h)后与对照比较实验组细胞增殖受到明显抑制(F=21.389、57.231、73.444,P<0.05)。160 nmol/L的Bufalin作用后细胞迁移数为(114.33±5.51),明显低于对照组(270.67±13.65,F=241.989,P<0.01)。CD133分子表达水平为(1.48±0.14),低于对照组(3.22±0.46,F=202.589,P<0.01),细胞凋亡率[(32.57±0.69)%]高于对照组[(16.35±0.33)%,F=171.288,P<0.01]。AMD3100组中,与对照组比较AMD310020μmol/L作用48 h细胞活性为(87.58±7.10)%,40μmol/L作用48 h为(67.84±0.75)%(F=57.231,P<0.05),40μmol/L作用72 h[(72.57±10.23)%,F=73.444,P<0.05]使细胞增殖明显受抑。40μmol/L的AMD3100作用后,与对照组细胞迁移个数[(270.67±13.65)个]明显低于实验组[(193.33±8.08)个,F=241.898,P<0.01]。CD133分子表达水平实验组为3.07±0.11,对照组为3.22±0.46,差异无统计学意义(F=202.589,P>0.05)。凋亡率Objective To investigate the effects of Bufalin,AMD3100 as a drug,alone or in combination,on cell proliferation,migration,apoptosis and CD133 protein expression after acting on gallbladder cancer cell line(GBC-SD).Methods Totally,20 specimens of wax blocks of gallbladder cancer surgically resected at our hospital from January 2017 to December 2021 were collected,and the differences in the expression levels of CD133 in tumor tissues as well as in paracancerous tissues were detected by immunohistochemical techniques.GBC-SD cells were treated with different concentrations of Bufalin(80,160,320 nmol/L);AMD3100(10,20,40μmol/L)and the combination(Bufalin 160 nmol/L+AMD310020μmol/L).Cell counting kit-8(CCK-8)method was used to observe the effect of drugs on tumor cell proliferation.Transwell method was used to detect tumor cell migration ability.Annexin V/propidium iodide(PI)method was used to detect apoptosis,and flow cytometry was used to detect CD133 expression.Rates were compared byχ2 test,and one-way and two-way ANOVAs were used for comparisons between groups.Results The positive expression rate of CD133 molecules in human gallbladder cancer tissues was 85%,which was significantly higher than that of 30%in paraneoplastic tissues.The expression of CD133 molecules was localized in the cytoplasm and nucleus(χ2=10.23,P<0.01).The cell proliferation of the experimental group was significantly inhibited by Bufalin(80,160,320 nmol/L)after 24,48,72 h as compared with the control(F=21.389,57.231,73.444,P<0.05).The number of migrating cells after treatment of Bufalin with 160 nmol/L was(114.33±5.51),which was significantly reduced compared to the control group(270.67±13.65,F=241.989,P<0.01).The expression level of CD133 molecules was(1.48±0.14)was lower in Bufalin group than that in the control group(3.22±0.46,F=202.589,P<0.01),and the apoptosis rate(32.57±0.69)%was elevated compared to the control group[(16.35±0.33)%,F=171.288,P<0.01].In the AMD3100 group,cell proliferation was inhibited in 20μmol/L at 48 h[(87.58

关 键 词：胆囊癌 CD133 AMD3100 BUFALIN 

分 类 号：R735.8[医药卫生—肿瘤]