沙蟾毒精对骨肉瘤细胞系143B的抗肿瘤作用及安全性  

Antitumor effect and safety of arenobufagin on osteosarcoma cell line 143B

作　　者：方硕 鲁蓉[1] 刘艳 FANG Shuo;LU Rong;LIU Yan(Wuhan Railway Vocational College of Technology,Wuhan 430074,China/Orthopedic Department,Renmin Hospital of Wuhan University,Wuhan 430060,China;不详)

机构地区：[1]武汉铁路职业技术学院,武汉市430074 [2]武汉大学人民医院骨科,武汉市430060

出　　处：《临床合理用药》2025年第6期26-30,共5页Chinese Journal of Clinical Rational Drug Use

基　　金：武汉铁路职业技术学院招标科研项目(YZB202307)。

摘　　要：目的探讨沙蟾毒精对骨肉瘤细胞系143B的抗肿瘤作用及安全性。方法分别用0、75、150 nM沙蟾毒精处理骨肉瘤细胞系143B细胞24 h,通过细胞毒性实验、菌落形成实验评价沙蟾毒精对143B细胞的抑制作用,Hoechst 33258染色法、流式细胞术检测细胞凋亡情况,Transwell细胞侵袭实验检测143B侵袭和转移能力,Western blotting检测细胞蛋白表达情况,小鼠体内毒性实验检测药物毒性。结果CCK-8实验结果显示,143B细胞生长均受到抑制。随着时间和浓度的增加,细胞生长抑制率逐渐升高。体外实验结果显示,处理组细胞菌落形成少于对照组(P<0.01)。Hoechst 33258染色法结果显示,与对照组相比,处理组细胞核更固缩、碎裂、细胞凋亡率高于对照组(P<0.01)。此外,裂解caspase-3/9和促凋亡蛋白Bax表达水平升高,抗凋亡蛋白Bcl-2表达水平逐渐下降。Transwell细胞侵袭实验结果显示,处理组侵袭性迁移细胞数量明显减少(P<0.01)。此外,处理组MMP-2/MMP-9蛋白表达水平低于对照组。小鼠体内毒性实验结果显示,未见小鼠死亡。3组血尿素氮、血肌酐、丙氨酸氨基转移酶、天冬氨酸氨基转移酶水平比较,差异无统计学意义(P>0.05)。高剂量组乳酸脱氢酶和肌酸激酶同工酶水平高于对照组(P<0.01)。结论沙蟾毒精能够抑制骨肉瘤细胞增殖,诱导细胞凋亡,抑制骨肉瘤细胞的侵袭和转移,但沙蟾毒精具有心脏毒性。Objective To investigate the antitumor effect and safety of arenobufagin on osteosarcoma cell line 143B.Methods Osteosarcoma cell line 143B cells were treated with 0,75,and 150 nM arenobufagin for 24 hours.Inhibitory action of arenobufagin on osteosarcoma cell line 143B was evaluated through cytotoxicity experiment and colony formation assays.Cell apoptosis was detected using Hoechst 33258 staining and flow cytometry.The invasion and metastasis ability of 143B cells was assessed by Transwell cell invasion experiment.Western blotting was used to detect cellular protein expression,and in vivo toxicity experiment in mice were conducted to assess drug toxicity.Results CCK-8 assay results showed that,the growth of 143B cells was inhibited.With increasing time and concentration,the cell growth inhibition rate gradually increased.In vitro experimental results indicated that,the number of colonies formed in the treatment group was less than that in the control group(P<0.01).Hoechst 33258 staining results showed that,compared with those in the control group,the nuclei of the treatment group were more pycnotic and fragmented,the apoptosis rate of cells in the treatment group was higher than that in the control group(P<0.01).Additionally,the expression levels of cleaved caspase-3/9 and the pro-apoptotic protein Bax increased,while the expression level of the anti-apoptotic protein Bcl-2 gradually decreased.Transwell cell invasion experiment results showed a significant reduction in the number of invasive and migratory cells in the treatment group.Furthermore,the expression levels of MMP-2/MMP-9 proteins in the treatment group were lower than those in the control group.In vivo toxicity experiment results in mice showed no mortality.There were no statistically significant difference in BUN,SCr,ALT,AST levels among the three groups(P>0.05).However,LDH and CK-MB levels in the high-dose group were higher than those in the control group(P<0.01).Conclusion Arenobufagin can inhibit the proliferation of osteosarcoma cell line 143B c

关 键 词：骨肉瘤 骨肉瘤细胞系143B 沙蟾毒精 细胞凋亡 侵袭与转移 

分 类 号：R73[医药卫生—肿瘤]