量子点标记与绿色荧光素标记对survivin反义寡核苷酸生物功能影响的比较

目的 观察量子点和绿色荧光素分别标记的survivin反义寡核苷酸(ASODN)转染人乳腺癌细胞株MCF-7后,在标记存在时间、survivin的表达及细胞增殖、凋亡方面有无差异.方法 将量子点和绿色荧光素分别标记的survivin ASODN通过脂质体转染MCF-7,MTT法检测细胞生长抑制率,RT-PCR检测survivin mRNA,Western blot检测survivin蛋白表达,流式细胞仪分析细胞凋亡率变化,荧光倒置显微镜观察细胞内荧光物质分布.结果 量子点与绿色荧光素分别标记的survivin ASODN作用MCF-7后,survivin mRNA和survivin蛋白表达均下降,但两者间无显著差异(P＞0.05).两者细胞生长抑制率、凋亡率间无显著差异(P＞0.05).转染后4 d绿色荧光素标记组细胞内荧光消失,而量子点标记组荧光1周仍存在.结论 量子点标记survivin ASODN与绿色荧光素标记比较,对survivin的表达及细胞增殖、凋亡方面无差异,而标记更稳定,存在时间更长.

Biological activity of survivin antisense oligonucleotide labeled with quantum dots or green fluorescein: a comparative study

Objective To compare the durability of quantum dots with that of green fluorescein for labeling survivin antisense oligonucleotide(ASODN)and investigate the difference in growth and apoptosis of cells transfected with the labeled survivin ASODN.Methods Survivin ASODN labeled with quantum dots or green fluorescein was transfected into MCF-7 cells via LipolifectmainTM2000.The proliferation of MCF-7 cells was assessed with MTT assay,survivin mRNA expression determined by RT-PCR and its protein expression measured by Western blot analysis.The apoptosis rate of the transfected cells was estimated by flow cytometry,and the fluorescence distribution in the cells observed under fluorescent inverted microscope.Results The mRNA and protein expressions of survivin were significantly decreased in the MCF-7 cells after cell transfection with survivin ASODN labeled with quantum dots or green fluorescein,and no significant difference was noted between the two labeling methods(P>0.05).Nor did survivin ASODN transfection with different labeling methods produced significant difference in cell proliferation and apoptotic rate(P>0.05).For green fiuorescein labeling,the fluorescence disappeared 4 days after transfection,whereas the fluorescence sustained for 1 week for quantum dots labeling.Conclusion Survivin ASODNs labeled with quantum dots and green fiuorescein do not significantly differ in survivin expression or the transfected cell proliferation and apoptosis rate,but quantum dot labeling can be more stable with longer maintcnance of the labeling.