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-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，转守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IJTEP-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-2mRNA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 transfeeted 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. Conelusion The sil0 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.