Effect of β-estradiol on the expression of human MGMT gene in cells in vitro

Z. Nidoieva, L. Lukash, A. P. Yatsyshyna
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Abstract

To determine whether the steroid hormone β-estradiol affects the transcription of the human MGMT gene. The expression status of the DNA repair enzyme O (6)-methylguanine-DNA methyltransferase (MGMT) is important when planning the treatment of patients with oncology, because its presence in tumor cells may reduce the effectiveness of alkylating chemotherapy. Alkylating chemotherapy is combined with hormone therapy in the treatment of tumors, so the study of the effect of β-estradiol on the number of transcripts of the human MGMT gene in cells in vitro has not only theoretical and also practical significance. Methods. We used a variety of methods, including culture, molecular genetic and biochemical, such as RNA isolation, cDNA synthesis, reverse transcriptase PCR, agarose and polyacrylamide gel electrophoresis, and statistical processing of the results. Results. We found a tendency of β-estradiol to downregulate the MGMT gene at mRNA level in both HEp-2 and 293 cells in a range of concentrations, except concentrations 1 and 5 nmol/L of β-estradiol in 293 cells. Conclusions. Our data support the hypothesis that β-estradiol is one of the hormonal regulators of the MGMT gene.
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β-雌二醇对人MGMT基因体外表达的影响
确定类固醇激素β-雌二醇是否影响人MGMT基因的转录。DNA修复酶O(6)-甲基鸟嘌呤-DNA甲基转移酶(MGMT)的表达状态在规划肿瘤患者的治疗时很重要,因为它在肿瘤细胞中的存在可能会降低烷基化化疗的有效性。在肿瘤的治疗中,烷基化化疗与激素治疗相结合,因此研究β-雌二醇对体外细胞中人MGMT基因转录本数量的影响不仅具有理论意义,而且具有实际意义。方法。我们使用了多种方法,包括培养、分子遗传和生化,如RNA分离、cDNA合成、逆转录酶PCR、琼脂糖和聚丙烯酰胺凝胶电泳,并对结果进行统计处理。结果。我们发现,除了293细胞中β-雌二醇浓度为1和5 nmol/L外,HEp-2和293细胞中β-雌二醇在一定浓度范围内均有下调MGMT基因mRNA水平的趋势。结论。我们的数据支持β-雌二醇是MGMT基因的激素调节因子之一的假设。
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