红花黄色素通过活性氧影响TGF-β1诱导的卵巢癌SKOV-3细胞生物学特性的机制学研究

Mechanism Study of Safflor Yellow Affecting the Biological Characteristics of Ovarian Cancer SKOV-3 Cells Induced by TGF-β1 through Reactive Oxygen Species

目的:探讨红花黄色素对转化生长因子-β1(TGF-β1)诱导的卵巢癌SKOV-3细胞增殖、侵袭和迁移的影响及其机制。方法:采用噻唑蓝法检测不同浓度(0.15,0.25,0.50,1.00 mg·L^(-1))红花黄色素对SKOV-3细胞增殖活力的影响,并根据红花黄色素抑制作用结果将SKOV-3细胞分为对照组、红花黄色素组(0.25 mg·L^(-1))、TGF-β1组(10 ng·ml^(-1))和红花黄色素+TGF-β1组(0.25 mg·L^(-1)+10 ng·ml^(-1)),采用噻唑蓝法、Transwell小室实验和划痕实验分别检测各组细胞增殖、侵袭和迁移能力,免疫印迹法检测各组细胞中E-钙黏素、N-钙黏蛋白和波形蛋白的表达,活性氧检测试剂盒检测各组细胞内活性氧水平。结果:红花黄色素可呈浓度依赖性抑制SKVO3细胞增殖。与对照组相比,10 ng·ml^(-1)TGF-β1处理后SKVO3细胞增殖、侵袭、迁移能力和细胞中N-钙黏蛋白、波形蛋白表达水平以及细胞内活性氧水平明显增强(P<0.05),且细胞中E-钙黏素蛋白表达水平明显降低(P<0.05);而0.25 mg·L^(-1)红花黄色素处理后SKVO3细胞则与TGF-β1处理后的结果相反(P<0.05);与TGF-β1组相比,红花黄色素联合TGF-β1可明显降低TGF-β1对SKVO3细胞的影响(P<0.05)。结论:红花黄色素可抑制TGF-β1诱导的卵巢癌SKOV-3细胞增殖、侵袭和迁移,其作用机制可能与抑制细胞内活性氧水平,减弱其介导的上皮间质转化有关。

Objective: To investigate the effects of safflower yellow on the proliferation,invasion and migration of ovarian cancer SKOV-3 cells induced by TGF-β1 and underlying mechanism. Methods: The effects of safflower yellow(0.15,0.25,0.50,1.00 mg·L^(-1)) on the proliferation of SKOV-3 cells were detected by MTT method. According to the results of safflower yellow inhibition,SKOV-3 cells were divided into control group,safflower yellow group(0. 25 mg· L-1),TGF-β1 group(10 ng·ml^(-1)) and safflower yellow + TGF-β1 group(0. 25 mg· L-1+ 10 ng· ml-1). The cell proliferation,invasion and migration was detected by MTT method,Transwell chamber test and scratch test,respectively. The expressions of E-cadherin,N-cadherin and vimentin in the cells of each group were detected by Western blot,and reactive oxygen species level was detected by oxygen species kit. Results: Safflower yellow inhibited the proliferation of SKVO3 cells in a concentration-dependent manner. Compared with those in the control group,the abilities of proliferation,invasion and migration,the expressions of N-cadherin and vimentin proteins,and the level of intracellular reactive oxygen species of SKVO3 cells treated with 10 ng·mL-1 TGF-β1 were significantly increased(P<0.05),and the expression of E-cadherin protein in SKVO3 cells was significantly decreased(P<0.05),while SKVO3 cells treated with 0.25 mg·L^(-1) safflower yellow were contrary to those treated with TGF-β1(P<0.05). Compared with TGF-β1 group,the combined effects of safflower yellow and TGF-β1 could significantly reduce the effects of TGF-β1 on SKVO3 cells(P<0.05). Conclusion: Safflower yellow can inhibit the proliferation,invasion and migration of ovarian cancer SKOV-3 cells induced by TGF-β1,and its mechanism may be related to the inhibition of intracellular reactive oxygen species level and the subsequent attenuation of oxygen species-mediated epithelial-mesenchymal transformation.