Si-10靶点对不同COX-2蛋白表达状态肺癌细胞体外恶性增殖的影响

Effects of Si-10 on malignant proliferation of lung cancer cells with different COX-2 expression states in vitro

目的:研究Si-10靶点对不同环氧化酶-2(COX-2)蛋白表达状态肺癌细胞干扰效果及对肺癌细胞恶性增殖的影响。方法:将psi-10及空载体(pEGFP)分别转染到不同COX-2表达状态的肺癌细胞,建立转染细胞株。采用反转录聚合酶链反应和蛋白质印迹方法检测COX-2表达水平的变化。通过细胞生长曲线、集落形成验研究这一靶点对肺癌细胞体外恶性增殖的影响。结果:转染后24、48和72h,转空载体的细胞均有绿色荧光表达,转染psi-10的801D、A549和LTEP-A2细胞均未观察到绿色荧光。在A549和LTEP-A2细胞中COX-2表达均受到抑制,但抑制效果不同,与A549细胞比较Si-10对LTEP-A2细胞COX-2表达抑制更为明显。与其母系比较,LTEP-A2-10和A549-10细胞COX-2mR-NA表达量分别降低64.2%和61.2%;COX-2蛋白表达量分别降低60.2%和56.2%。LTEP-A2-10和A549-10细胞生长明显减慢,集落形成率减少。与A549-10细胞比较,LTEP-A2-10细胞生长更慢,集落形成率更低,集落更小。Si-10靶点对COX-2不表达的801D细胞生长没有抑制作用。结论:si10对不同COX-2基因表达的肺癌细胞有不同的干扰效果,而不同干扰的效果对细胞的恶性增殖有着不同的抑制作用。

OBJECTIVE：To explore the effects of Si-10 on the lung cancer cells with different cyclooxygenase-2 （COX-2） expression states by using the same target to interfere in COX-2 gene expression and study the effect on the malignant proliferation of these cells in vitro.METHODS：The siRNA expression vector （psi-10） and the vacant vector （pEGFP） were transfected into these cells with different COX-2 expression states respectively and the transfected cell strains were constructed.The change of COX-2 expression levels was examined by Western blot and RT-PCR.The effects on the proliferation of lung cancer cells were studied by the cell growth curve and clonogenic assay.RESULTS：These cells transfected pEGFP had the expression of GFP and 801D-10,A549-10 and LTEP-A2-10 had not in 24,48 and 72 hours after transfected.The expression of COX-2 was inhibited in LTEP-A2 and A549 expressing COX-2 but the inhibited effects were different.The expression of COX-2 was inhibited more obviously in LTEP-A2 cells than in A549 cells.In contrast to their maternal lines,the levels of COX-2 mRNA of LTEP-A2-10 and A549-10 cells reduced by 64.2% and 61.2% respectively and these of COX-2 protein reduced by 60.2% and 56.2% respectively.The growth of LTEP-A2-10 and A549-10 cells slowed and the clonal formation rate reduced.The growth of 801D-10 cells had not obvious changes.CONCLUSION：The si10 has different interfering effects on lung cancer cells with different COX-2 expression states and the different interfering results have different effects on cell malignant proliferation.