【摘要】 目的观察丹参酮ⅡA(TanⅡA)对宫颈癌鳞癌细胞SiHa细胞侵袭的抑制作用及诱发细胞发生凋亡情况,探讨其可能作用机制。方法常规体外培养人宫颈鳞癌癌细胞系SiHa,加入0.5、1.0、2.0、5.0 mg/L浓度的TanⅡA,继续培养24 h。通过倒置显微镜观察细胞形态学变化,采用MTT试验检测细胞增殖情况,细胞划痕实验检测TanⅡA对细胞迁移能力的影响,用Transwell小室法检测TanⅡA对SiHa细胞侵袭的抑制作用,激光共聚焦显微镜观察SiHa细胞凋亡变化。Western blotting法检测MMP-2、NF-κB p65蛋白的表达。结果细胞形态学观察显示,不同浓度TanⅡA均可导致细胞形态学发生改变;MTT试验表明,TanⅡA对SiHa细胞的生长有明显抑制作用,且抑制作用随药物浓度的增加而增强(P<0.05,P<0.01);划痕实验结果显示,随TanⅡA药物浓度的增加,SiHa细胞迁移能力下降(P<0.01);Transwell小室实验显示,TanⅡA对SiHa细胞体外的侵袭力具有很强的抑制作用;Western blotting结果表明,TanⅡA给药能够抑制SiHa细胞MMP-2蛋白的表达与NF-κB p65的激活(P<0.01)。结论 TanⅡA能够抑制人宫颈鳞癌细胞SiHa细胞侵袭与诱导凋亡,其机制可能与抑制NF-κB蛋白的激活下调MMP-2的表达有关。更多还原

【Abstract】 Objective To observe the effects of inhibiting invasion and inducing apoptosis of tanshinone ⅡA( TanⅡA) in human cervical squamous carcinoma SiHa cells,and discuss its possible mechanism. Methods The human cervical squamous carcinoma SiHa cell lines were routinely cultured in vitro and treated with TanⅡA at concentration of 0. 5,1. 0,2. 0 and 5. 0 mg/L respectively. Then the cells were cultured for 24 hours. The morphological changes of the cells were observed by inverted microscope. The cell proliferation was detected by MTT assay,the effect of TanⅡA on cell migration was detected by cell scratch test,the inhibitory effect of TanⅡA on SiHa cell invasion was detected by Transwell test,and the change of SiHa cell apoptosis was observed by laser scanning confocal microscope. The protein expressions of MMP-2 and NF-κB p65 were detected by Western blotting. Results The morphological observation showed that tanshinoneⅡA with different concentrations could all lead to the changes of cell morphology. MTT assay indicated that tanshinone ⅡA showed the obviously inhibitory effect on SiHa cells growth,and the inhibitory effect was increased with the increasing drug concentration( P < 0. 05,P < 0. 01). The result of scratch test showed that the SiHa cell migration was decreased with the increasing TanⅡA concentration( P < 0. 01). The Transwell test suggested that TanⅡA showed strong inhibitory effect on SiHa cell invasion in vitro. The results of Western blotting showed that TanⅡA could inhibit the protein expression of MMP-2 and activation of NF-κB p65 in SiHa cells( P < 0. 01). Conclusion TanⅡA can inhibit the invasion and induce the apoptosis in human cervical squamous carcinoma SiHa cells,and its possible mechanism may be associated with inhibiting NF-κB activation and down-regulating MMP-2 expression.更多还原