摘要
目的:探讨蛋白激酶C(PKC)抑制剂星形孢菌素(STS)对肺腺癌A549细胞侵袭移动的影响及作用机制。方法:取A549细胞加入不同浓度STS(1、10、100 nmol.L-1)处理后,应用Transwell小室检测A549细胞移动抑制率和侵袭抑制率;免疫荧光染色法检测细胞内蛋白水解酶基质金属蛋白水解酶-9(MMP-9)和尿激酶型纤溶酶原激活剂(uPA)蛋白表达;激光共聚焦法观测细胞内骨架蛋白微管蛋白α-tubulin的分布,Western blot法检测胞膜和胞浆内PKC-α活性。同时设立不加STS处理的对照组进行比较。结果:与对照组比较,A549细胞的移动抑制率、侵袭抑制率、MMP-9和uPA蛋白表达量均明显下降(P<0.05或P<0.01);胞浆内α-tubulin蛋白分布不均;胞膜内PKC-α蛋白含量减少,而胞浆内PKC-α蛋白含量无明显变化。结论:PKC抑制剂STS能抑制A549细胞移动和侵袭;其机制可能与抑制PKC-α的活性,减弱肿瘤细胞中MMP-9、uPA表达,诱导肿瘤细胞骨架变化相关。
OBJECTIVE：To evaluate the effects of protein kinase C（PKC） inhibitor staurosporine（STS） on cell invasion ability of lung adenocarcinoma cell line A549 cells and its mechanism.METHODS：Transwell assay was applied to observe the motility inhibition rate and invasion inhibition rate of A549 cells after A549 cells were treated with STS at different concentrations（1,10,100 nmol·L^-1）.Protein expression of matrix metalloproteinase-9（MMP-9） and urokinase-type plasminogen activator（uPA） were detected by immunofluoresence stain assay.Laser confocal scanning microscopy was used to observe the distribution of α-tubulin.Western blot was used to determine the activity of PKC-α in membrane and endochylema.Control group were not treated with STS.RESULTS：Compared with control group,motility inhibition rate and invasion inhibition rate of A549 cells and protein expression of MMP-9 and uPA were decreased significantly（P〈0.05 or P〈0.01）.The distribution of α-tubulin in endochylema showed uneven situation.The content of PKC-α in the membrane was decreased while that in endochylema had no obvious change.CONCLUSION：PKC inhibitor STS could inhibit the motility and invasion of A549 cells.The inhibition effect may be associated with inhibiting the activity of PKC-α,decreasing protein expression of MMP-9 and uPA and inducing cytoskeleton change.
出处
《中国药房》
CAS
CSCD
北大核心
2010年第25期2336-2339,共4页
China Pharmacy
基金
湖北省卫生厅科研基金资助项目(JX2B89)
