樟脑对MRC-5细胞p53 mRNA表达的影响  

Effect of camphor on expression level of p53 mRNA in MRC-5 cells

作　　者：刘晓燕[1] 段承刚[1] 唐小军 蔡仕钰[1] 郑波[1] 罗映[1] 赖国超[1] 申淑琬[1] 于海清[1] 

机构地区：[1]泸州医学院医学基础研究中心 [2]泸州医学院附属医院心胸外科,四川泸州646000

出　　处：《泸州医学院学报》2013年第2期114-116,共3页Journal of Luzhou Medical College

基　　金：国家自然科学基金项目(81172237);四川省教育厅项目(12ZA241);四川省卫生厅项目(120379)

摘　　要：目的:检测樟脑(C10H16O)处理MRC-5细胞对p53基因mRNA表达水平的影响。方法:DMEM培养基培养MRC-5细胞,设试验组和对照组。1×105细胞/孔的细胞密度接种于6孔板,每组设三复孔,180μg.ml-1的樟脑分别作用MRC-5细胞24h和48h。MTT法测定细胞相对增殖百分率。提取细胞总RNA,逆转录得cDNA。GAPDH mRNA作为内参,p53/GAPDH mRNA为相对表达量,反转录半定量聚合酶链式反应(SQ-PCR)分析MRC-5细胞中p53表达水平,试验重复三次。结果:180μg.ml-1樟脑作用MRC-5细胞24h和48h后,细胞相对增殖百分率均高于90%。给药24h,对照组和试验组MRC-5细胞p53 mRNA表达水平分别为0.81±0.24和0.56±0.13(P>0.05)。给药48h,对照组和试验组MRC-5细胞p53 mRNA表达水平分别为0.76±0.11和1.08±0.19(P>0.05)。给药24h和48h,试验组MRC-5细胞p53 mRNA表达水平差异有统计学意义(P<0.05)。结论:180μg.ml-1浓度的樟脑对MRC-5为安全给药浓度。给药后MRC-5细胞p53 mRNA表达水平上调有统计学意义,提示樟脑可能间接影响p53基因的表达调控。Objective： To investigate the p53 mRNA expression level in MRC-5 cells after treated with camphor （CloHx60）. Methods： MRC-5 cells were cultured by using DMEM and divided into the camphor group and the control group, lxl0s cells/well were seeded evenly in a six-well plate,three wells for each group. MRC-5 cells were treated with 180μg·ml-1 camphor for 24h and 48h. The relative growth rate was measured via MTT method. Total RNA was extracted and cDNA was synthesized via reverse transcription. With GAPDH mRNA as reference, the relative quantity of p53/GAPDH mRNA expression level was analyzed by using semi-quantitative polymerase chain reaction （SQ-PCR）. The experiment was repeated three times. Results： After treated with 180μg·ml-1 camphor, the relative growth rate of MRC-5 cells was greater than 90% both at 24h and 48h. The p53 mRNA expression levels in control group and camphor group were 0.81±0.24 and 0.56±0.13 at 24h （/9〉0.05） and 0.76_±0.11 and 1.08±0.19 at 48h （P〉0.05）. The up-regulation of mRNA expression level at 48h was significantly higher than that at 24h in camphor group（P〈0.05）. Conclusion： The 180μg·ml-1 camphor was safe to MRC-5 cells. In comparison with the expression level at 24h after treated with camphor, the p53 mRNA is upregulated more significantly at 48h, which indicates that camphor may affect the regulation of p53 expression indirectly.

关 键 词：樟脑 MRC-5 P53 

分 类 号：R774[医药卫生—眼科]