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目的探讨佛波酯(phorbol 12-myristate 13-acetatefor,PMA)对人单核白血病细胞THP-1中颗粒溶素(granulysin,GLS)表达的激活作用。方法分别用不同浓度的PMA(130、160、180 nmol/L)诱导THP-1细胞24 h,RT-PCR法检测细胞中GLS基因mRNA的转录,筛选PMA最佳诱导浓度;用PMA最佳诱导浓度分别诱导THP-1细胞12、24和48 h,RT-PCR法检测GLS基因mRNA的转录,筛选最佳诱导时间;用PMA最佳诱导浓度分别诱导THP-1细胞12、24、48和72 h,Western blot法检测GLS蛋白的表达,免疫细胞化学法检测48 h时GLS蛋白的表达。结果以160 nmol/L的PMA诱导THP-1细胞24 h,细胞中GLS基因mRNA的转录水平最高;诱导48 h后可检测到GLS蛋白大量表达。结论 PMA可激活THP-1细胞中GLS的表达,本实验为后续进一步研究GLS表达的调控机制奠定了基础。
Objective To investigate the effect of phorbol 12-myristate 13-acetatefor (PMA) on the expression of granulysin (GLS) in human monocytic leukemia cell line THP-1. Methods THP-1 cells were induced by different concentrations of PMA (130,160,180 nmol / L) for 24 h, respectively. The transcription of GLS mRNA was detected by RT-PCR and the optimum concentration of PMA was screened. THP-1 cells were induced at different concentrations for 12, 24 and 48 h, RT-PCR was used to detect the transcription of GLS mRNA, and the optimal induction time was screened. THP-1 cells were induced by PMA at optimal concentrations of 12,24,48 and 72 h. The expression of GLS protein was detected by Western blot. The expression of GLS protein was detected by immunocytochemistry at 48 h. Results THP-1 cells were induced by PMA at 160 nmol / L for 24 h, and the transcription level of GLS mRNA was the highest at 48 h. A large amount of GLS protein was detected at 48 h after induction. Conclusions PMA can activate GLS expression in THP-1 cells. This study lays the foundation for further study on the regulatory mechanism of GLS expression.