细胞周期素D1反义脱氧寡核苷酸对A549细胞增殖及凋亡的调控作用

提要:目的探讨脂质体转染细胞周期素D1(cyclinD1)反义脱氧寡核苷酸链(antisenseoligodeoxynucleotide,ASON)对A549细胞增殖及细胞周期的影响。方法采用脂质体转染技术将cyclinD1ASON转入A549细胞培养,用MTT法检测细胞生长状况,应用流式细胞术(FACS)、检测细胞DNA梯形带等方法观察细胞凋亡及RT PCR法检测cyclinD1蛋白的表达。结果脂质体转染cyclinD1ASON组和单纯cyclinD1ASON组的A549细胞存活率较对照组显著降低(P<0.01)。脂质体转染cyclinD1ASON组cyclinD1表达水平(32%)较对照组(83%)明显减低(P<0.01),并能检测出DNA断裂。结论脂质体转染cyclinD1ASON能在体外明显抑制肺腺癌细胞的生长,且有诱导细胞凋亡、抑制细胞内cyclin D1合成的作用,可能会成为基因治疗的一个新靶点。

Effect of liposomal transfection of cyclin D1 antisense oligodeoxynucleotide on A549 cell proliferation and apoptosis

Objective To explore the effect of liposomal transfection of cyclin D1 antisense oligodeoxynucleotide (ASON) on A549 cell proliferation and apoptosis. Methods By liposomal transfection technique, cyclin D1 ASON was co-cultured with A549 cells. The cell growth curve was determined by MTT assay. The protein and mRNA of cyclin D1 were measured by FACS and RT-PCR. Results In the cyclin D1 ASON liposomal transfection group, the proliferation of A549 cells was significantly inhibited as compared to that in cyclin D1 ASON group (P<0.01). The expression of cyclin D1 in the cyclin D1 ASON liposomal transfection group was significantly lower than that in the control group (32% vs 83%, P<0.01), and the DNA ladder and apoptosis body was displayed. Conclusion Liposomal transfection of cyclin D1 ASON can inhibit in vitro the proliferation of lung cancer cells, which might provide a new target for gene therapy.