CAJ | 학술논문

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M G63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100 ng／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w nt3a的蛋白刺激浓度呈非正相关关系，在各个浓度中，以100 ng／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）。100 ng／mL的wnt3a蛋白刺激在培养6 h时β‐catenin基因表达最高，并且在6 h前，β‐catenin基因表达会随时间延长而增加。6 h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。
목적：분석실시형광정량 PCR기술（FQ‐PCR）진단인골육류 MG63세포계중β‐catenin기인표체。방법배양인골육류M G63세포계，대기진행분조，대조조세포사용10％적태우혈청이급함유청매소화련매소적DEME고당배양기배양。이자격조칙재차기출상용100 ng／mL적wnt3a세포진행배양。분별재제0、1、2、3、4천분별채집3개공적세포，채용세포소화적방식계산세포개수，채용FQ‐PCR검측인골육류MG63세포계중β‐catenin기인표체。결과재세포생장적제0、1천，량조MG세포수량대비차이무통계학의의（P＞0．05）；이재제2、3、4천량조적세포개수대비차이유통계학의의（ P＜0．05）。β‐catenin기인표체여w nt3a적단백자격농도정비정상관관계，재각개농도중，이100 ng／mL적wnt3a단백자격β‐catenin기인표체최고，기타궤개농도적β‐catenin기인표체대비차이무통계학의의（P＞0．05）。100 ng／mL적wnt3a단백자격재배양6 h시β‐catenin기인표체최고，병차재6 h전，β‐catenin기인표체회수시간연장이증가。6 h후β‐catenin기인표체대비차이무통계학의의（P＞0．05）。결론FQ‐PCR능구반영인골육류MG63세포중β‐catenin기인표체。
Objective To analyze the real time FQ‐PCR technique for diagnosing β‐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 wnt3a cell culture .The cells in 3 holes were collected on 0 ,1 ,2 ,3 ,4 d respectively 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 .Results On 0 ,1 d of the cell growth ,the MG cells number had no statistical difference between the two groups ;while which on 2 ,3 ,4 d had 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 100 ng/mL wnt3a protein was highest ,the differences in theβ‐catenin gene ex‐pression had no statistical difference among other concentrations(P>0 .05) .Theβ‐catenin gene expression stimulated by 100 ng/mL wnt3a protein for 6 h culture reached highest ,moreover before 6 h ,theβ‐catenin gene expression was increased with time lapse .Theβ‐catenin gene expression after 6 h had no statistical difference(P>0 .05) .Conclusion FQ‐PCR can reflect theβ‐catenin gene expression in human osteosarcoma MG63 cells .