Objective: We investigated the relationship between CYP17A1-mediated DNA demethylation and proliferation, invasion, and metastasis of glioma cells.
Methods: The expression of CYP17A1 mRNA and protein in cells was determined by PCR and Western blot assays. The methylation status of CYP17A1 was detected by the MSP method. Cell proliferation and apoptosis were detected by MTT assays and flow cytometry. Cell invasion and metastasis were measured by cell invasion assays.
Results: The relative expression of CYP17A1 mRNA was significantly different among the model, experimental, and normal groups (P < 0.05). Relative expression was significantly decreased in the experimental group relative to the cancer model group (P < 0.05). Immunohistochemistry showed that expression of CYP17A1 in glioma was significantly higher than in the normal group (P < 0.05). Methylation analysis showed that CYP17A1 was not detected in normal cells, and the methylation rate in the model group was 89.03%. The methylation rate in the experimental group was 43.93%, which was significantly lower than that of the model group (P < 0.05). MTT assays showed that DHEA plus temozolomide (TMZ) pretreatment significantly inhibited cell proliferation rate (P < 0.05). Flow cytometry showed that DHEA plus TMZ pretreatment significantly increased apoptosis rate (P < 0.05). In colony formation assays, the number of CYP17A1 colonies in the model and experimental groups was 78.09% ± 10.21% and 38.97% ± 11.32%, respectively. The number of colonies in the experimental group was significantly lower than in the model group (P < 0.05). The migration ability of the model group was significantly higher than that of the control group (P < 0.05). The invasion rate of the experimental group was significantly lower than that of the model group (P < 0.05).
Conclusion: CYP17A1-induced DNA demethylation can inhibit proliferation, invasion, and metastasis of glioma cells.

中文翻译：

---

CYP17A1介导的DNA去甲基化与胶质瘤细胞增殖、侵袭和转移的关系

目的：我们研究了 CYP17A1 介导的 DNA 去甲基化与胶质瘤细胞增殖、侵袭和转移之间的关系。
方法：采用PCR和Western blot检测细胞内CYP17A1 mRNA和蛋白的表达。MSP法检测CYP17A1甲基化状态。MTT法和流式细胞术检测细胞增殖和凋亡。通过细胞侵袭测定法测量细胞侵袭和转移。
结果： CYP17A1 mRNA相对表达量在模型组、实验组和正常组之间差异有统计学意义（P < 0.05）。实验组相对于癌模型组的相对表达显着降低（P < 0.05)。免疫组化显示CYP17A1在胶质瘤中的表达显着高于正常组（P < 0.05）。甲基化分析显示正常细胞中未检测到CYP17A1，模型组甲基化率为89.03%。实验组甲基化率为43.93%，显着低于模型组（P < 0.05）。MTT检测显示DHEA加替莫唑胺（TMZ）预处理显着抑制细胞增殖率（P < 0.05）。流式细胞术显示DHEA加TMZ预处理显着增加细胞凋亡率（P< 0.05)。在集落形成实验中，模型组和实验组的CYP17A1集落数分别为78.09%±10.21%和38.97%±11.32%。实验组菌落数显着低于模型组（P < 0.05）。模型组迁移能力明显高于对照组（P < 0.05）。实验组侵袭率显着低于模型组（P < 0.05）。
结论： CYP17A1诱导的DNA去甲基化可抑制胶质瘤细胞的增殖、侵袭和转移。