1Department of Biology, Kazerun Branch, Islamic Azad University, Kazerun, Islamic Republic of Iran.
2Department of Biology, Hamedan Branch, Islamic Azad University, Hamedan, Islamic Republic of Iran.
Physiol Int. 2020 Oct 9;107(3):406-418. doi: 10.1556/2060.2020.00034. Print 2020 Oct 17.
Progesterone has been reported to inhibit the proliferation of breast cancer and osteosarcoma cells; however, its inhibitory mechanism has not yet been clarified. The aim of the present study was to clarify the effects of progesterone on apoptosis in breast cancer (MCF-7) and human osteosarcoma (MG-63) cells.
In this experimental study the cytotoxic effect of progesterone was measured in MCF-7 and MG-63 cells exposed to different concentrations of progesterone using MTT assay, and effective concentrations were identified. The expression levels of the Bax, P53 and Bcl-2 genes were evaluated by real-time PCR, and caspase-3, 8 and 9 activity levels were determined using a colorimetric method. Hoechst staining and flow cytometry were used to confirm apoptosis. The data were statistically analyzed using one-way analysis of variance (ANOVA) and independent-samples t-test.
Compared to the control group, we observed a significant increase in the expression levels of the Bax and P53 genes and the activity levels of caspase-3 and 9, and a significant decrease in the expression level of the Bcl-2 gene in MCF-7 and MG-63 treated with effective concentration of progesterone. The caspase-8 activity level did not change significantly in treated MG-63 but increased in treated MCF-7 cells. Hoechst staining and flow cytometry results confirmed apoptosis in the cells exposed to effective concentration of progesterone.
The cytotoxic effect of progesterone on breast cancer and osteosarcoma cells was mediated by apoptotic pathways. In this context, progesterone triggers the extrinsic and intrinsic apoptotic pathways in MCF-7 cells and induces the intrinsic apoptotic pathway in MG-63 cells.
有报道称孕激素可抑制乳腺癌和骨肉瘤细胞的增殖，但孕激素的抑制机制尚不清楚。本研究旨在阐明孕激素对乳腺癌（MCF-7）和人骨肉瘤（MG-63）细胞凋亡的影响。
在这项实验研究中，通过 MTT 法测量了不同浓度孕激素暴露下 MCF-7 和 MG-63 细胞中的细胞毒性效应，并确定了有效浓度。通过实时 PCR 评估 Bax、P53 和 Bcl-2 基因的表达水平，并通过比色法测定 caspase-3、8 和 9 的活性水平。使用 Hoechst 染色和流式细胞术确认细胞凋亡。使用单因素方差分析（ANOVA）和独立样本 t 检验对数据进行统计学分析。
与对照组相比，我们观察到在接受有效浓度孕激素处理的 MCF-7 和 MG-63 中，Bax 和 P53 基因的表达水平以及 caspase-3 和 9 的活性水平显著增加，Bcl-2 基因的表达水平显著降低。处理后的 MG-63 中的 caspase-8 活性水平没有显著变化，但处理后的 MCF-7 细胞中的 caspase-8 活性水平增加。Hoechst 染色和流式细胞术结果证实了细胞暴露于有效浓度孕激素后的凋亡。
孕激素对乳腺癌和骨肉瘤细胞的细胞毒性作用是通过凋亡途径介导的。在这种情况下，孕激素在 MCF-7 细胞中触发外源性和内源性凋亡途径，并在 MG-63 细胞中诱导内源性凋亡途径。
