熊果酸对喉癌Hep-2细胞侵袭的影响及机制研究

The impact and mechanisms of ursolic acid on Hep-2 cells invasion

目的探讨熊果酸对人喉癌Hep-2细胞侵袭能力的影响及机制。方法不同浓度熊果酸处理人喉癌Hep-2细胞不同时间后,采用MTT法检测细胞的增殖活性,Boyden chamber检测细胞的侵袭能力,Western blot检测周期相关基因NF-κB蛋白表达的变化。明胶酶谱检测细胞培养上清液基质金属蛋白酶(MMP)-9和MMP-2活性的变化。结果熊果酸对人喉癌Hep-2细胞的生长具有明显的增殖抑制作用,其作用表现为剂量依赖性和时间依赖性(P<0.01)。Boyden chamber结果显示以熊果酸处理Hep-2细胞后,侵袭能力呈浓度依赖性下降(P<0.01)。Western blot结果显示熊果酸对人喉癌Hep-2细胞NF-κB蛋白表达呈浓度依赖性下调(P<0.01)。明胶酶谱结果显示熊果酸对人喉癌Hep-2细胞MMP-9和MMP-2活性呈浓度依赖性下调(P<0.01)。结论熊果酸通过降低MMP-9和MMP-2活性及NF-κB蛋白表达来抑制人喉癌Hep-2细胞的增殖和侵袭。

Objective To study the effect of ursolic acid on human laryngeal carcinoma Hep-2cells invasion and to explore its mechanism.Methods Methabenzthiazuron（MTT）was used to observe the proliferation of Hep-2cells treated with various concentrations of ursolic acid at different time.Boyden chamber was used to compare ursolic acid effect on cell invasion.Western blot was used to the measure the NF-κB protein expression.Gelatin zymography was used to the activity of MMP-9and MMP-2.Results Ursolic acid inhibited human laryngeal carcinoma Hep-2cells growth,its effect showed dose dependent and time dependent（P〈0.01）.Boyden chamber results revealed that after the ursolic acid treatment Hep-2cells,invasion ability showed a concentration dependent decreased（P〈0.01）.Western blot results revealed that ursolic acid showed a concentration dependent decreased on human laryngeal Hep-2cells NF-κB protein expression（P〈0.01）.Gelatinases spectrum revealed that ursolic acid a concentration dependent decreased on human laryngeal Hep-2cells MMP-9and MMP-2cells activity（P〈0.01）.Conclusion Ursolic acid inhibit human laryngeal cancer Hep-2cell proliferation and invasion,its mechanism is related to down-regulate MMP-9and MMP-2activity and the NF-κB protein expression.