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目的研究Survivin反义寡核苷酸对A549细胞增殖和凋亡的影响。方法针对Survivin mRNA序列设计了反义寡核甘酸,转染A549细胞,PT-PCR检测细胞中survivin基因的mRNA含量;Western印迹显示Survivin蛋白水平;以MTT法检测细胞增殖,TUNEL法检测细胞凋亡。结果Survivin反义寡核苷酸可明显降低A549细胞中survivin的mRNA和蛋白水平。MTF比色法结果说明人Survivin反义寡核苷酸抑制A549细胞增殖,抑制率远高于无义寡核苷酸组和空白对照组。TUNEL法检测结果显示,反义寡核苷酸还显著增强A549细胞对抗肿瘤药高三尖杉酯碱的敏感性,在较低高三尖杉酯碱浓度下,反义寡核苷酸转染细胞的凋亡率明显高于其它对照组。结论sur- vivin反义寡核苷酸有望应用于肿瘤的辅助治疗之中。
Objective To study the effect of Survivin antisense oligonucleotide on the proliferation and apoptosis of A549 cells. Methods Antisense oligodeoxynucleotides were designed according to the sequence of Survivin mRNA. A549 cells were transfected with A549 cells. The mRNA level of survivin gene was detected by PT-PCR. Survivin protein was detected by Western blotting. Cell proliferation was detected by MTT assay and apoptosis was detected by TUNEL assay . Results Survivin antisense oligonucleotide significantly decreased the mRNA and protein level of survivin in A549 cells. MTT colorimetric results showed that human Survivin antisense oligonucleotide inhibited A549 cell proliferation, the inhibition rate was much higher than the non-sense oligonucleotide group and the blank control group. TUNEL assay showed that antisense oligonucleotides also significantly increased the sensitivity of A549 cells to the antineoplastic homoharringtonine, and at lower concentrations of homoharringtonine, antisense oligonucleotides transfected cells The apoptosis rate was significantly higher than other control groups. Conclusion Survivin antisense oligonucleotides are expected to be used in the adjuvant therapy of tumors.