人绒毛膜癌细胞JEG-3中SEPSECS、TRNAU1AP蛋白表达稳定性的分析

Analyze the Stability of Half-Life of SEPSECS and TRNAU1AP in Choriocarinoma Cell Line on JEG-3

目的:通过测定人绒毛膜癌细胞JEG-3中SEPSECS、TRNAU1AP两蛋白的半衰期,分析JEG-3细胞内SEPSECS、TRNAU1-AP两蛋白表达的稳定性。方法:采用蛋白合成抑制剂放线菌酮作用于人绒毛膜癌细胞JEG-3,用MTT法确定放线菌酮对细胞的最适作用浓度,之后用最适浓度的放线菌酮处理JEG-3,利用Western blot检测10个时间点SEPSECS、TRNAU1AP蛋白表达量,确定SEPSECS、TRNAU1AP蛋白半衰期。结果:CHX作用于细胞的最适浓度为0.1μg/mL,与对照组相比,SEPSECS蛋白表达量在4 h、8 h有显著性差异(P<0.01);而TRNAU1AP蛋白表达量在2 h、4 h有显著性差异(P<0.01)。结论:在JEG-3细胞中SEPSECS蛋白的半衰期可能为4 h～8 h,而TRNAU1AP蛋白的半衰期可能为2 h～4 h,前者半衰期明显大于后者,由此,可推测SEPSECS蛋白在JEG-3细胞中表达的稳定性要高于TRNAU1AP蛋白,同时也为进一步分析二者在其他肿瘤细胞的表达情况提供依据。

Objective：To analyze the stability of SEPSECS and TRANU1AP in JEG-3 cell by testing the half-life of these proteins.Methods：Protein synthesis inhibitor Cycloheximide（CHX） wes used to handle JEG-3 and the optimum effect concentration of CHX was measured by MTT.Western bolt was performed to detect 10 different timepoints.Eventually,the half-life of SEPSECS and TRNAU1AP in JEG-3 cell was determined.Results：Optimal concentration of CHX on JEG-3 is 0.1 μg/mL（P0.01）.Compared with control,4 h and 8 h have significant differences on SEPSECS（P0.01）.Similarly,2 h and 4 h have significant differences on TRNAU1AP（P0.01）.Conclusion：Half-life of SEPSECS may be between 4 h and 8 h,while,the half-life of TRNAU1AP which between 2 h and 4 h is shorter than SEPSECS obviously.Therefore,it indicates that SEPSECS has a higer possession of stability than TRNAU1AP.