白藜芦醇阻断转化生长因子-/Smad通路抑制人结肠癌细胞系SW480侵袭转移实验研究  

Resveratrol Inhibited the Metastasis of Colon Cancer SW480 Cells Via Suppressing the TGF-/Smad Pathway

作　　者：宋康杰 

机构地区：[1]浙江省宁波市北仑区妇幼保健院药剂科,宁波315800

出　　处：《浙江中西医结合杂志》2016年第1期27-30,共4页Zhejiang Journal of Integrated Traditional Chinese and Western Medicine

摘　　要：目的探讨中药白藜芦醇对SW480细胞活力及侵袭转移力的影响。方法将人结肠癌细胞系SW480用白藜芦醇处理48h,MTT法检测白藜芦醇对SW480细胞活力的影响;流式细胞术检测白藜芦醇对SW480细胞周期的影响;Transwell穿膜试验检测白藜芦醇对SW480细胞侵袭转移能力的影响;western blot实验检测肿瘤细胞转移相关蛋白E-上皮型细胞钙黏蛋白(E-cadherin)、波形蛋白(Vimentin)及基质金属蛋白酶-9(MMP-9)的表达水平;Western blot实验检测上游磷酸化Smad蛋白(p-Smad)的表达水平。结果对照组及5、10、15、20、40μmol/L白藜芦醇组对SW480细胞活力的抑制率分别为0、(5.4±1.0)%、(22.5±3.3)%、(36.4±4.4)%、(62.2±6.1)%、(85.1±7.5)%,各组间细胞活力抑制率差异有统计学意义(P<0.05)。对照组及5、10、15、20、40μmol/L白藜芦醇组对SW480细胞周期G2/M阻滞率分别为(2.3±0.3)%、(4.3±0.5)%、(8.9±1.2)%、(15.6±2.3)%、(22.7±2.7)%、(30.4±2.9)%,各组间G2/M阻滞率差异有统计学意义(P<0.05)。对照组及转化生长因子-β(TGF-β)组、TGF-β+5μmol/L白藜芦醇组、TGF-β+10μmol/L白藜芦醇组、TGF-β+15μmol/L白藜芦醇组、TGF-β+20μmol/L白藜芦醇组、TGF-β+40μmol/L白藜芦醇组穿过Transwell膜的SW480细胞数分别为(204.2±19.4)个、(453.5±41.6)个、(411.2±45.2)个、(315.7±32.7)个、(246.3±27.2)个、(197.8±23.5)个、(97.4±11.2)个,各组间细胞数差异有统计学意义(P<0.05)。白藜芦醇体外处理置于TGF-β培养体系中的SW480细胞后,E-cadherin表达水平显著上升,Vimentin及MMP-9表达水平显著下降(P<0.05)。白藜芦醇显著降低TGF-β诱导的SW480细胞Smad蛋白磷酸化水平。结论白藜芦醇有良好的体外抗结肠癌的生物活性,更重要的是它能通过阻断TGF-β/Smad通路抑制结肠癌细胞的侵袭转移。Objective To investigate the effect of resveratrol on the cell viability and metastasis in SW480 cells in vitro. Methods SW480 cells were treated with resveratrol for 48h： the cell viability was measured by MTT assay, the cell cycle was evaluated by flow cytometry analysis, and the metastasis was evaluated by Transwell assay.Metastasis associated proteins E-cadherin, vimentin and MMP-9 in SW480 cells treated with resveratrol were detected by using Western Blot analysis. The upstream pathway proteinphosphorylated Smad（p-Smad） was detected in SW480 cells treated with resveratrol by Western Blot analysis. Results Cell viability inhibitory rate of SW480 cells in control group, 5μmol/L resveratrol group, 10μmol/L resveratrol group, 15μmol/L resveratrol group, 20μmol/L resveratrol group, and 40μmol/L resveratrol group were 0,（5.4±1.0）%,（22.5±3.3）%,（36.4±4.4）%,（62.2±6.1）%,and（85.1±7.5）%, respectively, with a significant difference among them（P〈0.05）. The cell cycle G2/M inhibitory rate of SW480 cells in above groups were as follows：（2.3 ±0.3）%,（4.3 ±0.5）%,（8.9 ±1.2）%,（15.6 ±2.3）%,（22.7 ±2.7）%,and（30.4±2.9）%, with a significant differenceamong groups（P〈0.05）. The number of cells thatpenetrated the Transwell membrane in control group, TGF-β group, TGF-β＋5μmol/L resveratrol group, TGF-β＋10μmol/L resveratrol group, TGF-β＋15μmol/L resveratrol group, TGF-β＋20μmol/L resveratrol group, and TGF-β＋40μmol/L resveratrol group were as follows： 204.2 ±19.4, 453.5 ±41.6, 411.2 ±45.2, 315.7 ±32.7, 246.3 ±27.2, 197.8 ±23.5, 97.4 ±11.2,with a significant differenceamong groups（P〈0.05）. Western blot analysis showed the expression of E-cadherin was significantly up-regulated, while the Vimentin and MMP-9 were down-regulated by resveratrol treatment in SW480cells（all P〈0.05）. The expression ofp-Smad was significantly down-regulated when SW480 cells were treated with resveratrol. Conclusion Resveratrol showed a significan

关 键 词：结肠癌细胞 SW480 白藜芦醇 转移 TGF-β SMAD 

分 类 号：R735.35[医药卫生—肿瘤]