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目的:探讨丹参酮ⅡA(TanⅡA)对人卵巢癌细胞SKOV3的生长抑制和诱导凋亡作用及其可能机制。方法:培养人卵巢癌细胞SKOV3,经TanⅡA作用后,通过四甲基偶氮唑蓝(MTT)比色法测定丹参酮ⅡA对人卵巢癌细胞SKOV3的增殖抑制作用;流式细胞术检测细胞周期;RT-PCR检测Survivin与Smac/DIABLO mRNA表达水平;免疫荧光细胞化学检测Survivin蛋白及Smac/DIABLO蛋白的表达。结果:1.0～10.0μg/ml TanⅡA对人卵巢癌细胞SKOV3具有生长抑制作用,并具有时间-剂量依赖性。实验组较对照组G1/G2期细胞数量增多,而S期细胞数量减少(P<0.05)。随着TanⅡA干预浓度的增加和作用时间的延长,Survivin mRNA的表达受到明显抑制(P<0.05),而Smac/DIABLO mRNA的表达明显上调(P<0.05)。Survivin蛋白及Smac/DIABLO蛋白均表达于SKOV3细胞,其表达多定位于细胞质中,偶见于细胞核。结论:TanⅡA对人卵巢癌细胞SKOV3具有生长抑制和诱导凋亡作用,其作用机制可能是通过抑制人卵巢癌细胞SKOV3 Survivin和促进Smac/DIABLO的表达而实现的。
Objective: To investigate the effect of TanⅡA on the growth and apoptosis of human ovarian cancer cell line SKOV3 and its possible mechanism. Methods: The human ovarian cancer cell line SKOV3 was cultured and treated with TanⅡA, the proliferation of human ovarian cancer cell line SKOV3 was determined by MTT assay. The cell cycle was detected by flow cytometry. The expression of Survivin and Smac / DIABLO mRNA was detected by RT-PCR. Survivin protein and Smac / DIABLO protein expression were detected by immunofluorescence cytochemistry. Results: TanⅡA at a concentration of 1.0 ~ 10.0μg / ml could inhibit the growth of human ovarian cancer cell line SKOV3 in a time-and dose-dependent manner. Compared with the control group, the number of cells in G1 / G2 phase increased and the number of S phase cells decreased (P <0.05). The expression of Survivin mRNA was significantly inhibited (P <0.05) and the expression of Smac / DIABLO mRNA was significantly upregulated (P <0.05) with the increase of TanⅡA concentration and the prolongation of action time. Survivin protein and Smac / DIABLO protein were expressed in SKOV3 cells, the expression of more localized in the cytoplasm, occasionally in the nucleus. Conclusion: TanⅡA can inhibit the growth and induce the apoptosis of human ovarian cancer cell line SKOV3. The mechanism may be through inhibiting the expression of SKOV3 and promoting the expression of Smac / DIABLO in human ovarian cancer cell line SKOV3.