高压电场不可逆电穿孔抑制人肝癌耐药细胞BEL-7402/5-氟尿嘧啶生长的作用  

Study on irreversible electroporation inhibited the proliferation of 5-fluorouracil resistant liver cancer cells BEL-7402/5-fluorouracil

作　　者：董健[1,2] 赵诗迪 徐碧翌 柯梦云 吕毅[1] Dong Jian;Zhao Shidi;Xu Biyi;Ke Mengyun;Lyu Yi(National Local Joint Engineering Research Center for Precision Surgery&Regenerative Medicine,Shaanxi Provincial Center for Regenerative Medicine and Surgical Engineering,the First Affiliated Hospital of Xi’an Jiaotong University,Xi’an 710061,China;Department of Vascular Surgery,the First Affiliated Hospital of Xi’an Jiaotong University,Xi’an 710061,China)

机构地区：[1]西安交通大学第一附属医院精准外科与再生医学国家地方联合工程研究中心陕西省再生医学与外科工程研究中心,710061 [2]西安交通大学第一附属医院血管外科,710061

出　　处：《中华实验外科杂志》2020年第10期1790-1792,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金(81727802);国家重点研发计划(2018YFC0115300);2019年西安交通大学本科生科研训练和实践创新基金项目(XJ201910698123)。

摘　　要：目的:观察不可逆电穿孔技术对5-氟尿嘧啶(5-Fu)耐药的人肝癌细胞(BEL-7402/5-Fu,购自南京凯基生物科技发展有限公司)生长、细胞膜完整性以及荷瘤鼠肿瘤生长的影响。方法:细胞计数试剂盒(CCK-8)检测细胞存活率。透射电镜、扫描电镜拍照以及乳酸脱氢酶(LDH)检测来评价细胞膜完整性。构建BEL-7402/5-Fu细胞裸鼠异种移植瘤模型来评价体内抗肿瘤效果。采用单因素方差分析。结果:高压电场显著抑制BEL-7402/5-Fu细胞生长并且呈场强依赖性,高压电场处理24 h后,500、750、1000、1500、2000 V/cm组的细胞存活率为(89.56±12.37)%、(66.29±7.47)%、(29.19±3.62)%、(17.66±1.64)%、(11.74±1.08)%(F=77.730,P<0.01)。扫描电镜以及透射电镜结果表明,在场强为1000 V/cm时,实验组的细胞膜完整性以及细胞器结构损伤严重。高压电场500、750、1000、1500、2000 V/cm各组的细胞培养基上清中LDH分泌水平分别为(35.33±8.74)、(64.33±7.67)、(144.02±12.16)、(194.33±5.13)、(207.33±11.01)、(213.33±20.81)U/L(F=122.690,P<0.01),呈场强依赖性。高压电场能够抑制BEL-7402/5-Fu荷瘤鼠肿瘤的生长,模型组的瘤重为(1.51±0.32)g,高压电场治疗组小鼠的瘤重为(0.28±0.05)g(F=619.870,P<0.01),抑瘤率达81.45%。结论:高压电脉冲能显著抑制BEL-7402/5-Fu细胞生长。Objective To investigate the effect of irreversible electroporation(IRE)on the growth of human 5-fluorouracil resistant liver cancer cell line BEL-7402/5-fluorouracil(5-Fu).Methods Cell counting kit-8(CCK-8)assay was used to detect cell viability.Cell membrane integritywas detected via scanning electron microscopy and transmission electron microscopy and measuring lactic dehydrogenase(LDH)levels.In vivo experiment was conducted by establishing subcutaneous transplanted tumors in nude mice.The statistical method used one-way analysis of variance.Results The IRE technology significantly inhibited the growth BEL-7402/5-Fu cells.The cell viability was(89.56±12.37)%,(66.29±7.47)%,(29.19±3.62)%,(17.66±1.64)%,(11.74±1.08)%in the electric field of 500,750,1000,1500,2000 V/cm,respectively(F=77.730,P<0.01).In the 1000 V/cm group,the integrity of the cell membrane and the structure of organelles was seriously damage.LDH secretion in the supernatant of cell culture medium was(35.33±8.74),(64.33±7.67),(144.02±12.16),(194.33±5.13),(207.33±11.01),(213.33±20.81)U/L(F=122.690,P<0.01)in the 500,750,1000,1500,2000 V/cm treatment group,respectively.IRE prevented the tumor growth in vivo.The tumor weight was(1.51±0.32)g and(0.28±0.05)g(F=619.87,P<0.01)in the model group and high voltage pulse treated group,respectively.Conclusion High voltage electric pulse significantly inhibited the growth of BEL-7402/5-Fu cells and would be a promising approach in cancer treatment.

关 键 词：高压电场 5-氟尿嘧啶耐药肝癌 生长 

分 类 号：R735.7[医药卫生—肿瘤]