生存素反义寡核苷酸对人恶性黑素瘤A375细胞生物学行为的影响

Effect of Survivin Antisense Oligonucleotide on Biological behaviors of Human Malignant Melanoma A375 Cells

目的研究生存素反义寡核苷酸(AS-ODN)经脂质体介导转染对A375细胞增殖、凋亡的影响。方法合成生存素硫代反义寡核苷酸(AS-ODN),脂质体携带转染人恶性黑素瘤A375细胞。采用台盼蓝计数、软琼脂克隆形成试验、电镜、流式细胞仪检测AS-ODN对A375细胞增殖及凋亡的影响。通过裸鼠致瘤实验观察AS-ODN对A375细胞动物体内增殖的抑制作用。结果生存素反义寡核苷酸能降低G2/M期细胞百分比,诱导细胞凋亡发生,明显抑制A375细胞的生长和克隆形成,裸鼠体内A375细胞恶性增殖潜力下降。结论脂质体介导生存素AS-ODN能有效抑制A375细胞增殖、诱导肿瘤细胞凋亡。

Objective To investigate the effects of liposome-mediated survivin antisense oligonucleotide on proliferation and apoptosis of huaman malignant melanoma A375 cells. Methods Survivin phosphorothioate antisense oligndeoxynucleotide （ AS-ODN） were synthesized and transfected into the target cells. Proliferation and apoptosis of A375 cells were measured by trypan blue staining,soft agar colony forming assay,electron microscope and flow cytometry. In nude xenograft of cancer, we investigated the tumorigenesis ability of the A375 cell tranfected with survivin antisense olignnucleotide in vivo. Results Transfection of AS-ODN markedly inhibited the proliferation of A375 cells and colony forming compared with M-ODN and S-ODN （ P 〈 0.05 ） . ASODN treatment resulted accumulation of apoptotic cells and coincident loss of mitotic （G2/M） cells. Malignant proliferative potential of A375 cells was inhibited in nude following transfection with AS-ODN. Conclusion Liposome - mediated Survivin AS - ODN can evidently inhibit cell proliferation and induce apoptosis of A375 cells. Survivin has a significance effect on human malignant melanoma cell division and apoptosis.