CAJ | 학술논문

目的改进Caspase-3/7活性检测方法.方法在顺铂诱导的HeLa细胞凋亡模型中,分别利用传统的和改良的实验方法检测Caspase-3/7的活性变化,并与Western印迹实验结果进行方法学比较.结果改良的实验方法显示不同剂量顺铂诱导下,HeLa细胞Caspase-3/7活性有剂量依赖性增高,与Western印迹实验结果相一致,但传统实验方法显示HeLa细胞Caspase-3/7活性呈现先增高后降低的趋势.结论由于参测细胞数不同,导致这种Caspase-3/7活性检测方法不能真实反映细胞凋亡程度.本研究成功建立了一种新的改良型Caspase-3/7活性检测方式,这种检测方法可以排除不同凋亡诱导方式诱导细胞凋亡时所产生的因参测细胞数不同所造成的Caspase-3/7活性检测的误差.
목적 개진Caspase-3/7활성검측방법.방법 재순박유도적HeLa세포조망모형중,분별이용전통적화개량적실험방법검측Caspase-3/7적활성변화,병여Western인적실험결과진행방법학비교.결과 개량적실험방법현시불동제량순박유도하,HeLa세포Caspase-3/7활성유제량의뢰성증고,여Western인적실험결과상일치,단전통실험방법현시HeLa세포Caspase-3/7활성정현선증고후강저적추세.결론 유우삼측세포수불동,도치저충Caspase-3/7활성검측방법불능진실반영세포조망정도.본연구성공건립료일충신적개량형Caspase-3/7활성검측방식,저충검측방법가이배제불동조망유도방식유도세포조망시소산생적인삼측세포수불동소조성적Caspase-3/7활성검측적오차.
Objective To improve the accuracy of Caspase-3/7 activity assay.Methods In cisplatin induced apoptotic Hela cell model,Caspase-3/7 activity of Hela cells was determined by traditional method or modified method.Meanwhile,cleaved PARP was detected by Western blot to determine apoptosis induction.Results The results of modified assay showed that Caspase-3/7 activity was increased in a cisplatin dose dependent manner,which was consistent with the Western blot result.However,the results of traditional assay showed that Caspase-3/7 activity of Hela cells was firstly increased but then declined,displaying a bias due to decreased cell numbers in the high dose of cisplatin.Conclusion Because the cell numbers involved in the assay were different,the traditional Caspase-3/7 activity assay is not reliable to reflect the apoptotic extent of tested samples.Our newly established Caspase-3/7 activity assay with internal control may correct the deviations of Caspase-3/7 activity assay caused by difference in cell numbers among samples,thus providing more accurate and reliable results.