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目的探讨细胞色素P450c17a酶(CYP17A1)在人神经胶质瘤细胞系T98G、U87和U251中的表达情况。方法采用Western blot和实时荧光定量PCR于蛋白质水平和mRNA水平检测细胞色素P450c17a酶(CYP17A1)在三种人胶质瘤细胞系中的表达。结果 Western blot检测出CYP17A1蛋白在胶质瘤细胞系T98G、U251和U87中的相对表达量为0.518±0.052、0.460±0.034和0.142±0.025。T98G和U251的CYP17A1蛋白表达水平明显高于U87,差异有统计学意义(P<0.05)。实时荧光定量PCR检测结果显示CYP17A1的mRNA在T98G、U251和U87中相对转录水平为1.000±0.122、0.960±0.079、0.611±0.045,T98G和U251细胞中CYP17A1的mRNA转录水平均高于U87,差异有统计学意义(P<0.05)。同时Western blot和实时荧光定量PCR都指出T98G和U251在CYP17A1在蛋白和mRNA的表达上无统计学差异(P>0.05)。结论人胶质瘤细胞系T98G和U251中CYP17A1的表达量较高,这两种细胞系可作为良好的细胞模型用来研究CYP17A1在胶质瘤中的作用机制以及以CYP17A1为靶点的肿瘤治疗等。
Objective To investigate the expression of cytochrome P450c17a (CYP17A1) in human glioma cell lines T98G, U87 and U251. Methods The expression of cytochrome P450c17a (CYP17A1) in three human glioma cell lines was detected by Western blot and real-time fluorescence quantitative PCR at the level of protein and mRNA. Results The relative expression levels of CYP17A1 protein in glioma cell lines T98G, U251 and U87 were 0.518 ± 0.052, 0.460 ± 0.034 and 0.142 ± 0.025 by Western blot. The expression of CYP17A1 in T98G and U251 was significantly higher than that in U87 (P <0.05). Real-time PCR results showed that the relative transcriptional levels of CYP17A1 mRNA in T98G, U251 and U87 were 1.000 ± 0.122, 0.960 ± 0.079 and 0.611 ± 0.045, respectively. The mRNA transcription levels of CYP17A1 in T98G and U251 cells were higher than those in U87 Statistical significance (P <0.05). Both Western blot and real-time PCR indicated that there was no significant difference in the expression of CYP17A1 protein and mRNA between T98G and U251 (P> 0.05). Conclusion The expression of CYP17A1 in human glioma cell lines T98G and U251 is high, and these two cell lines can be used as a good cell model to study the mechanism of action of CYP17A1 in glioma and tumor therapy targeting CYP17A1 Wait.