培美曲塞对SKP2蛋白表达的影响及意义

Impact of Pemetrexed on expression of SKP2 in adenocarcinoma of lung cell lines A549 cellandits significance

目的探讨S期激酶相关蛋白2(SKP2)在培美曲塞(PMX)处理后肺腺癌细胞A549中表达的改变及意义。方法根据PMX的浓度设1个对照组和4个实验组,分别为:0μmol/L组(A组)、0.01μmol/L组(B组)、0.1μmol/L组(C组)、1μmol/L组(D组)、10μmol/L组(E组),采用甲基噻唑基四唑(MTT)法检测A549细胞的增殖;利用流式细胞技术分析A549细胞周期的分布;通过Western blot法检测A549细胞SKP2蛋白表达。结果 PMX明显抑制A549细胞增殖,以10μmol/L、72h作用抑制率最高(P<0.01);与A组相比C、D、E组S期细胞显著增多(P<0.05);各组SKP2蛋白相对表达水平差异有统计学意义(P<0.01);与对照组相比,实验组SKP2蛋白相对表达水平显著增加(P<0.01)。结论 PMX能明显抑制A549细胞增殖且将其阻滞于S期,PMX处理后的A549细胞中SKP2蛋白表达上调,这可能影响PMX对肺腺癌的化疗疗效。

Objective To investigate the expression variation and significance of s-phase kinase-associated protein 2 （SKP2） in lung adenocareinoma A549 cells after the treatment of Pemetrexed（PMX）. Methods On the basis of the concentration of PMX,we set one control group and four experimental groups：0μM（group A） ,0.01μM（group B）, 0. 1μM（group C）, 1/aM（group D）, 10μM （group E）. The growth of A549 was tested by MTT assay. The effect of PMX on the cell cycle phase distribution of A549 was ana lyzed by flow cytometry. The expression of SKP2 protein of A549 cells was detected by Western blot. Results MTT assay showed that PMX could significantly inhibit the growth of A549 cells,and the highest inhibitory rate was（78.61 ± 3.66）%at the concen- tration of 10μM when A549 cells were cultured for 72 h（P〈0.01）. Meanwhile,S-phase cells in the group C,D and E was more than those in the group A（P〈0.05 ,P〈0.01 ,P〈0.01）. The difference of SKP protein expression among various groups was sig- nificant（P〈0.01） ,and compared with the control group, SKP2 protein expression was significantly increased in the experimental groups（P〈0.01）. Conclusion PMX could inhibit the growth of A549 cells and arrest them in S-phase. The expression of SKP2 protein in PMX-treated A549 cells is upregulated,which might influence the chemotherapy effect of PMX.