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目的观察乙酰丹参酮ⅡA(ATA)对人卵巢癌细胞增殖和凋亡的诱导作用及对雌激素诱发蛋白TFF1表达的影响,探讨可能的作用机制。方法通过MTT法计算ATA对卵巢癌细胞的活力以及IC50值;用流式细胞术检测凋亡率及细胞周期变化情况;用实时荧光定量聚合酶连反应(RT-PCR)检测TFF1 m RNA的表达水平,并通过构建双荧光素酶报告基因系统分析ATA对TFF1启动子活性的影响。结果 ATA对多种人卵巢癌细胞系均具有增殖抑制作用,对表达雌激素受体ER的肿瘤细胞效果最明显;能使SKOV3细胞周期阻滞在G2/M期,并诱导SKOV3细胞凋亡;能促进TFF1 m RNA的表达;能增强TFF1启动子的转录活性,且在TFF1启动子区域,AP-1结合位点对TFF1启动子的活性起绝对性作用。结论 ATA能有效抑制SKOV3的生长并可促进TFF1启动子的转录活性。
Objective To investigate the effect of acetyl tanshinone Ⅱ A (ATA) on the proliferation and apoptosis of human ovarian cancer cells and its effect on the expression of estrogen-induced protein TFF1, and to explore the possible mechanism. Methods The viability and IC50 of ATA on ovarian cancer cells were calculated by MTT assay. The apoptosis rate and cell cycle were detected by flow cytometry. The expression of TFF1 mRNA was detected by real-time fluorescence quantitative polymerase chain reaction (RT-PCR) Level and analyzed the effect of ATA on TFF1 promoter activity by constructing a dual luciferase reporter system. Results ATA could inhibit the proliferation of human ovarian cancer cell lines and had the most obvious effect on the tumor cells expressing estrogen receptor ER. It could arrest the cell cycle of SKOV3 in G2 / M phase and induce the apoptosis of SKOV3 cells. Can enhance the expression of TFF1 m RNA; enhance the transcriptional activity of the TFF1 promoter; and the absolute binding of the AP-1 binding site to the TFF1 promoter in the TFF1 promoter region. Conclusion ATA can effectively inhibit the growth of SKOV3 and promote the transcriptional activity of TFF1 promoter.