肺瘤平膏对共培养条件下A549细胞uPA、MMP-2、MMP-9mRNA及蛋白表达的影响

Effect of Feiliuping Ointment on uP A,MMP-2,MMP-9 mRNA and protein expression of A549 cell under co-culture conditions

目的肺瘤平膏对共培养条件下人肺腺癌细胞(A549细胞)尿激酶类纤溶酶(u PA)、基质金属蛋白酶-2(MMP-2)、MMP-9 mRNA及蛋白表达的影响。方法制备肺瘤平膏含药血清,建立A549细胞与巨噬细胞的共培养体系,通过荧光定量PCR、Western blot及ELISA法观察肺瘤平膏对共培养条件下u PA、MMP-2、MMP-9 mRNA及蛋白表达的影响。结果共培养条件下,A549细胞变成间充质细胞样,而加入肺瘤平膏含药血清组,部分A549细胞形态恢复上皮细胞样。共培养条件下A549细胞中MMP-2、MMP-9及u PA蛋白的编码基因PLAU的mRNA表达量显著升高(P<0.05),肺瘤平膏含药血清干预后,其表达量均显著降低(P<0.05);共培养条件下A549细胞中MMP-2、MMP-9蛋白表达量显著升高(P<0.05),肺瘤平膏含药血清干预后,其表达量均显著降低(P<0.05);共培养上清液中MMP-2、MMP-9、u PA蛋白浓度均随时间延长逐渐升高,肺瘤平膏可以降低24 h后上清液中u PA的浓度(P<0.05),而对MMP-2、MMP-9的浓度无明显影响(P>0.05)。结论肺瘤平膏可以降低共培养条件下A549细胞中MMP-2和MMP-9的含量及上清液中u PA的含量,但不能降低上清液中MMP-2和MMP-9的含量。

Objective To observe the effect of Feiliuping Ointment on uP A,MMP-2,MMP-9 mRNA and protein expression of A549 cell under co-culture conditions. Methods Feiliuping Ointment-containing serum（ FLP） was prepared; co-culture system for A549 cells and macrophage were established; the effects of Feiliuping Ointment on uP A,MMP-2,MMP-9 mRNA and protein expression of A549 cell under co-culture conditions were observed by RT-PCR,Western blot and ELISA. Results A549 cells were changed into mesenchymal-like cells in co-culture conditions,but some of them could recover to epithelial-like cell in FLP group; under co-culture conditions,the mRNA expression of PLAU of uP A,MMP-2,MMP-9 in A549 cell was obviously elevated（ P 0. 05）,but the mRNA expression were reduced by FLP intervention treatment（ P 0. 05）; under co-culture conditions,the expression of MMP-2,MMP-9in A549 cells was elevated（ P 0. 05）,but the expression of these protein levels by FLP intervention was reduced by intervention（ P 0. 05）; under co-culture conditions,MMP-2,MMP-9,uP A in the concentration were all increased with time,but after 24 hours,the concentration of uP A was reduced by FLP（ P 0. 05）,but MMP-2 and MMP-9 not affected（ P 0. 05）. Conclusion FLP Ointment can decrease the contents of MMP-2,MMP-9 in A549 cells and uP A in the concentration,but can not reduce the contents of MMP-2,MMP-9 in the concentration.