In cancer cells, the balance between proliferation and apoptosis is disturbed. There is a direct relationship between gene expression and the process of apoptosis. The two genes involved in apoptosis are Bax and Bcl-2, and it is now well established that some plant compounds can alter the expression of genes. The aim of this study is to determine the rate of change in the expression of these genes in the cell line MCF-7 treated with Dioscorea extract for 24, 48 and 72 h. For this purpose, the plant extract was prepared by Soxhlet method and diluted in different concentrations. MCF-7 and HFF cell lines were treated in three replicates with different concentrations of the extract at intervals of 24, 48, and 72 h. To evaluate the toxicity of the extract, the MTT assay was performed and the IC50 value was calculated. Both cell types were cultured at IC50 concentration with three treatments and three replicates. RNA extraction, cDNA synthesis and real-time PCR were then performed. Flow cytometry was performed to further confirm apoptosis. MTT results showed that 72 h treatment with Dioscorea extract in IC50 concentration had the greatest effect on the death of MCF-7 cancer cells, while the cells of the control cell line remained healthy. The results of the study of gene expression changes showed that when treated with the plant extract for 24 h, the increase in Bax gene expression and the decrease in Bcl-2 gene expression were not statistically significant. At 48-h treatment, the decrease in Bcl-2 expression was not statistically significant, whereas the increase in Bax expression, which was 2.1 times, was statistically significant. When treated with the plant extract for 72 h, Bax expression increased 2.72 times and Bcl-2 gene expression decreased 0.67 times. Flow cytometry showed that 72-h treatment with plant extract at a concentration of 438.35 µg/ml was the most effective treatment for MCF-7 cancer cell death. The expression ratio of Bax gene to Bcl-2 is equal to 4.06, which indicates the induction of more apoptosis by treatment with plant extract.

中文翻译：

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薯蓣提取物对 MCF-7 和 HFF 细胞系中 Bax 和 Bcl-2 基因表达的影响

在癌细胞中，增殖和凋亡之间的平衡被破坏。基因表达与细胞凋亡过程有直接关系。参与细胞凋亡的两个基因是 Bax 和 Bcl-2，现在已经确定一些植物化合物可以改变基因的表达。本研究的目的是确定用薯蓣提取物处理 24、48 和 72 小时的细胞系 MCF-7 中这些基因表达的变化率。为此，通过索氏提取法制备植物提取物并稀释成不同浓度。MCF-7 和 HFF 细胞系以 24、48 和 72 小时的间隔用不同浓度的提取物重复处理 3 次。为了评价提取物的毒性，进行MTT测定并计算IC50值。两种细胞类型均以 IC50 浓度培养，进行 3 次处理和 3 次重复。然后进行RNA提取、cDNA合成和实时PCR。进行流式细胞术以进一步确认细胞凋亡。MTT结果显示，IC50浓度的薯蓣提取物处理72小时对MCF-7癌细胞的死亡作用最大，而对照细胞系的细胞保持健康。基因表达变化研究结果显示，植物提取物处理24 h后，Bax基因表达量的增加和Bcl-2基因表达量的减少没有统计学意义。治疗48小时时，Bcl-2表达下降无统计学意义，而Bax表达增加2.1倍，有统计学意义。当用植物提取物处理72小时时，Bax表达增加2.72倍，Bcl-2基因表达减少0.67倍。流式细胞术显示，用浓度为 438.35 µg/ml 的植物提取物处理 72 小时是 MCF-7 癌细胞死亡的最有效治疗方法。Bax基因与Bcl-2的表达比等于4.06，这表明植物提取物处理诱导了更多的细胞凋亡。