摘要
目的分析实时荧光定量PCR技术(FQ-PCR)诊断人骨肉瘤MG63细胞系中β-catenin基因表达。方法培养人骨肉瘤MG63细胞系,对其进行分组,对照组细胞使用10%的胎牛血清以及含有青霉素和链霉素的DEME高糖培养基培养。而刺激组则在此基础上用100ng/mL的wnt3a细胞进行培养。分别在第0、1、2、3、4天分别采集3个孔的细胞,采用细胞消化的方式计算细胞个数,采用FQ-PCR检测人骨肉瘤MG63细胞系中β-catenin基因表达。结果在细胞生长的第0、1天,两组MG细胞数量对比差异无统计学意义(P>0.05);而在第2、3、4天两组的细胞个数对比差异有统计学意义(P<0.05)。β-catenin基因表达与wnt3a的蛋白刺激浓度呈非正相关关系,在各个浓度中,以100ng/mL的wnt3a蛋白刺激β-catenin基因表达最高,其他几个浓度的β-catenin基因表达对比差异无统计学意义(P>0.05)。100ng/mL的wnt3a蛋白刺激在培养6h时β-catenin基因表达最高,并且在6h前,β-catenin基因表达会随时间延长而增加。6h后β-catenin基因表达对比差异无统计学意义(P>0.05)。结论FQ-PCR能够反映人骨肉瘤MG63细胞中β-catenin基因表达。
Objective To analyze the real time FQ-PCR technique for diagnosingβ-catenin gene expression in human osteosarcoma MG63 cell line.Methods Human osteosarcoma cell line MG63 was cultured and grouped.The control group used 10% fetal bovine serum and the culture medium containing high glucose DEME penicillin and streptomycin.On this basis the stimulation group was cultured by using 100ng/mL wnt3 acell culture.The cells in 3holes were collected on 0,1,2,3,4drespectively and the cells number was counted by adopting the cell digestion way.FQ-PCR was adopted to detect theβ-catenin gene expression in human osteosarcoma cell line MG63.ResultsOn 0,1dof the cell growth,the MG cells number had no statistical difference between the two groups;while which on 2,3,4dhad statistically significant differences between the two groups（P〈0.05）.Theβ-catenin gene expression showed a non-positive correlation with the stimulation concentration of wnt3 protein,in various concentrations,theβ-catenin gene expression stimulated by 100ng/mL wnt3 aprotein was highest,the differences in theβ-catenin gene expression had no statistical difference among other concentrations（P〉0.05）.Theβ-catenin gene expression stimulated by 100ng/mL wnt3 aprotein for 6hculture reached highest,moreover before 6h,theβ-catenin gene expression was increased with time lapse.Theβ-catenin gene expression after 6hhad no statistical difference（P〉0.05）.Conclusion FQ-PCR can reflect theβ-catenin gene expression in human osteosarcoma MG63 cells.
出处
《检验医学与临床》
CAS
2015年第22期3307-3308,3311,共3页
Laboratory Medicine and Clinic
基金
福建省自然科学基金资助项目(2012J01410)
