CYP17A1在胶质瘤细胞系中表达及其抑制剂对胶质瘤细胞增殖的影响

发布时间：2019-01-05 17:11

【摘要】：目的:研究性激素合成通路中的关键酶之一的细胞色素P450c17a酶(CYP17A1)在三种人神经胶质瘤细胞系T98G、U87和U251中的表达水平,使用CYP17A1抑制剂Abiraterone干预胶质瘤细胞株T98G,并探索其对胶质瘤细胞增殖活性的影响,为涉及CYP17A1在胶质瘤中的作用机制以及以CYP17A1为潜在靶点的肿瘤治疗研究等方面奠定基础。方法:(1)选取细胞活性良好的三种人脑胶质瘤细胞系T98G、U87和U251为实验样本,采用Western blot和Real-time PCR的方法从蛋白质水平和m RNA水平来检测CYP17A1在三种胶质瘤细胞系T98G、U87和U251的表达水平,并比较CYP17A1在三种胶质瘤细胞系T98G、U87和U251间的蛋白质和m RNA表达水平差异。(2)通过CYP17A1抑制剂Abiraterone干预胶质瘤细胞株T98G,采用Real-time PCR,Western blot,cck8增殖实验及流式细胞术实验检测Abiraterone干预对T98G中CYP17A1表达水平、细胞增殖活性与细胞凋亡活性的影响。结果:(1)Western blot结果显示:检测出CYP17A1在三种人脑胶质瘤细胞系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)。人脑胶质瘤细胞系T98G和U251中CYP17A1表达水平均明显高于人脑胶质瘤细胞系U87,差异有统计学意义(P0.05)。Real-time PCR检测结果显示:CYP17A1的m RNA在三种人脑胶质瘤细胞系T98G、U251和U87中相对表达水平分别为(1.000±0.122)、(0.960±0.079)、(0.611±0.045)。其中,人脑胶质瘤细胞系T98G中CYP17A1的表达水平高于人脑胶质瘤细胞系U87,差异有统计学意义(P0.05);人脑胶质瘤细胞系U251中CYP17A1的表达水平也要高于人脑胶质瘤细胞系U87,差异同样具有统计学意义(P0.05)。但是,Western blot和Real-time PCR都指出人脑胶质瘤细胞系T98G和U251在CYP17A1在蛋白质和m RNA的表达差异无统计学意义(P0.05);(2)Abiraterone对胶质瘤的生物学活性产生抑制作用,能抑制胶质瘤细胞的增殖,促进胶质瘤细胞的凋亡,并且这种抑制及促进作用与Abiraterone的浓度相关(P0.05);(3)Abiraterone可以影响胶质瘤细胞的CYP17A1表达水平,影响的效果也与Abiraterone浓度相关(P0.05)。结论:(1)神经胶质瘤细胞系T98G、U87和U251均表达CYP17A1,其中,胶质瘤细胞系T98G和U251中CYP17A1的表达水平均明显高于人脑胶质瘤细胞系U87中CYP17A1的表达水平,但是在脑胶质瘤细胞系T98G和U251中CYP17A1的表达水平并无明显差异。(2)CYP17A1抑制剂能抑制胶质瘤细胞的增殖,促进胶质瘤细胞的凋亡,并影响胶质瘤细胞的CYP17A1表达水平。
[Abstract]:Aim: to study the expression of cytochrome P450c17a enzyme (CYP17A1), one of the key enzymes in sex hormone synthesis pathway, in three human glioma cell lines T98GnU87 and U251, and to use CYP17A1 inhibitor Abiraterone to interfere with the expression of cytochrome P450c17a enzyme (CYP17A1) in human glioma cell line T98G. The effects of CYP17A1 on the proliferation of glioma cells were explored, which laid a foundation for the mechanism of CYP17A1 in glioma and the study of tumor therapy with CYP17A1 as the potential target. Methods: (1) three kinds of glioma cell lines T98GU87 and U251 with good cell activity were selected as experimental samples. Western blot and Real-time PCR were used to detect CYP17A1 in three glioma cell lines T98G from protein level and m RNA level. The expression levels of U87 and U251 were compared between the three glioma cell lines T98GU87 and U251. (2) the CYP17A1 inhibitor Abiraterone was used to interfere with T98G and Real-time PCR,Western blot, was used to treat the glioma cell line T98G. Cck8 proliferation assay and flow cytometry were used to detect the effect of Abiraterone intervention on the expression of CYP17A1, cell proliferation activity and apoptosis activity in T98G. Results: (1) Western blot results showed that the relative expression of CYP17A1 was (0.518 卤0.052), () 0.460 卤0.034 and (0.142 卤0.025) in three human glioma cell lines T98GnU251 and U87. The expression of CYP17A1 in human glioma cell line T98G and U251 was significantly higher than that in human glioma cell line U87 (P0.05). The results of Real-time PCR detection showed that the expression of m RNA of CYP17A1 in three kinds of human glioma cell lines T98G was significantly higher than that in human glioma cell line U87 (P0.05). The relative expression levels of U251 and U87 were (1.000 卤0.122), (, 0.960 卤0.079), (, 0.611 卤0.045), respectively. The expression of CYP17A1 in human glioma cell line T98G was significantly higher than that in human glioma cell line U87 (P0.05). The expression of CYP17A1 in human glioma cell line U251 was also higher than that in human glioma cell line U87 (P0.05). However, both, Western blot and Real-time PCR indicated that there was no significant difference in the expression of T98G and U251 in CYP17A1 between protein and m RNA (P0.05). (2) Abiraterone inhibited the biological activity of glioma cells, inhibited the proliferation of glioma cells and promoted the apoptosis of glioma cells, and the inhibition and promotion effect was related to the concentration of Abiraterone (P0.05); (3) Abiraterone could affect the expression of CYP17A1 in glioma cells, and the effect was related to the concentration of Abiraterone (P0.05). Conclusion: (1) the expression of CYP17A1 in glioma cell line T98G and U251 was significantly higher than that in human glioma cell line U87 and U251, and the expression level of CYP17A1 in glioma cell line T98G and U251 was significantly higher than that in human glioma cell line U87. However, there was no significant difference in the expression of CYP17A1 between T98G and U251 glioma cells. (2) CYP17A1 inhibitor could inhibit the proliferation of glioma cells, promote the apoptosis of glioma cells, and affect the CYP17A1 expression level of glioma cells.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R739.41

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