反义寡核苷酸抑制MMP7基因表达对肺腺癌A549细胞凋亡敏感性影响的研究

目的观察基质溶解素(matrilysin,MMP7)反义寡核苷酸(antisense ligodeoxynucleotide,ASODN)对肺腺癌A549细胞凋亡敏感性的影响。方法采用硫代磷酸修饰的MMP7ASODN,通过脂质体导入A549细胞后,RT-PCR检测MMP7 ASODN对MMP7 mRNA表达的影响;流式细胞仪检测其对细胞膜Fas抗原表达率的影响;加入重组FasL,诱导凋亡,流式细胞仪检测凋亡率。结果RT-PCR结果表明,MMP7 ASODN转染能显著抑制A549细胞MMP7 mRNA表达。流式细胞仪检测表明,MMP7ASODN转染A549细胞后,与未转染组和错义寡核苷酸(scrambled oligodeoxynucleotide,SCODN)组比较,细胞膜Fas蛋白表达率增高,有显著性差异(P<0.01)。加入重组FasL,诱导细胞凋亡,细胞凋亡率明显增高,与未转染组和SCODN组有显著性差异(P<0.01)。结论MMP7ASODN可以抑制肺腺癌A549细胞株MMP7基因的表达,上调细胞膜Fas抗原表达,增强A549细胞凋亡敏感性。

Effect of MMP7 Antisense Oligodeoxynucleotide on Sensitivity of Apoptosis of Pulmonary Adenocarcinoma Cell Line A549

Objective To observe the sensitivity of apoptosis of pulmonary adenocarcinoma cell line A549 through inhibiting the expression of Matrilysin (MMP7)by MMP7 antisense oligodeoxynucleotide (ASODN). Methods After phosphorothioate MMP7 ASODN was transfected into A549 cells by liposome, the expressions of MMP7 were examined by RT-PCR, the expression of Fas was examined by Flow Cytometry (FCM), and the rate of apoptosis was examined by FCM after the FasL was added. Results After A549 cells were transfected with MMP7 ASODN, MMP7 mRNA expression was downregulated markedly. The expression of Fas was upregulated markedly (P<0.01).The rate of apoptosis increased as compared with A549 group and SCODN group after the FasL was added (P<0.01). Conclusion MMP7 ASODN can specifically inhibit the expression of MMP7 and enhance the expression of Fas and the sensitivity of apoptosis of A549 cells.