人胚胎干细胞源角质形成细胞用于药物毒理检测的探讨

Application of keratinocytes derived from human embryonic stem cells for toxicological evaluation of drugs

目的:探讨人胚胎干细胞源角质形成细胞(keratinocytes derived from human embryonic stem cells,K-hESCs)用于药物细胞毒理检测反应的可行性,为建立新的生物安全性评价模型提供依据.方法:应用CCK-8法检测细胞活性和细胞毒性.观察维A酸(RA)、5氟尿嘧啶(5-FU)、地塞米松(DEX)和青霉素G(PG)对K-hESCs、人原代牙龈上皮细胞(human gingival epithelial cells,HGECs)和人永生化口腔上皮细胞系(human immortalized oral epithelial cells,HIOEC)的细胞毒性反应.采用SPSS 16.0软件包对数据进行统计学分析.结果:RA作用于HGECs、HIOEC和K-hESCs 3种角质形成细胞后,其药物半数抑制浓度(IC50)分别为6.1±0.03、5.62±0.05和6.58±0.02;5-FU作用于3种细胞后,IC∞分别为1.65±0.02、3.00±0.02和1.72±0.04;DEX作用于3种细胞后,IC50分别为113.67±0.014、328±0.002和126.17±0.05;PG作用于3种细胞后,IC50分别为2200±1.34、3795±2.42和2880±1.5.4种药物对HIOEC和HGECs的IC50有统计学差异(P<0.05).对K-hESCs和HIOEC的IC50差异也有统计学意义(P<0.05),但对K-hESCs和HGECs的IC50差异无统计学意义(P>0.05).结论:在一般细胞毒性上,K-hESCs的药物半数抑制浓度比HIOEC更接近HGECs,可模拟人体正常细胞的反应.

PURPOSE:To preliminarily explore the cytotoxicological responses of keratinocytes derived from human embryonic stem cells(K-hESCs)to drugs,and to provide a basis for the establishment of a new biosafety evaluation model.METHODS:CCK-8 assay was used to detect cell viability and cytotoxicity.The detection of pharmacological response was observed and compared when K-hESCs directly derived from human embryonic stem cells,human gingival epithelial cells(HGECs),and human immortalized oral epithelial cells(HIOECs)were treated with retinoic acid(RA),5-fluorouracil(5-FU),dexamethasone(DEX),and penicillin G(PG).Statistical analysis was performed using SPSS 16.0 software package.RESULTS:After drugs were applied to HGECs,HIOEC and K-hESCs,the half inhibitory concentrations(IC50)of RA was 6.1±0.03,5.62±0.05 and 6.58±0.02,respectively.The IC50 of 5-FU was 1.65±0.02,3.00±0.02 and 1.72±0.04,respectively.The IC50 of DEX was 113.67±0.014,328±0.002 and 126.17±0.05,respectively.The IC50 of PG was 2200±1.34,3795±2.42 and 2880±1.5,respectively.The IC50 of the four drugs between HIOEC and HGECs had significant differences(P<0.05),but there was no significant difference in IC50 between K-hESCs and HGECs(P>0.05).The IC50 of K-hESCs and HIOEC also had significant differences(P<0.05).CONCLUSIONS:IC50 of K-hESCs was closer to HGECs than HIOEC.It was speculated that K-hESCs could simulate the response of normal human cells in cytotoxicity study.