摘要
目的以氨基脂为主要成分、叶酸受体为靶向的脂质体包裹MYCN基因小干扰性RNA,研究其体外对神经母细胞瘤LA-N-5细胞MYCN基因表达的抑制及细胞增殖的影响。方法制备氨基脂为主要成分、叶酸受体为靶向的脂质体包裹CY3标记的MYCNsiRNA转染LA-N-5细胞,以每微克蛋白中荧光强度半定量siRNA的转染效果;用定量RT-PCR检测转染前后MYCN基因mRNA变化;MTT法检测细胞增殖受抑情况。结果以100nmol/L终浓度转染LA-N-5细胞后每微克蛋白含siRNA(1808.5±140.2)pg,作用72hMYCN基因mRNA表达显著下降,抑制率79.2%,瘤细胞增殖受抑达66.2%。结论叶酸受体为靶向脂质体介导MYCNsiRNA在体外对LA-N-5细胞MYCNmRNA表达有显著抑制作用,明显抑制LA-N-5细胞增殖,为神经母细胞瘤靶向治疗、基因治疗开辟了新的思路。
Objective To study the inhibitory effect of folate receptor-targeted MYCN gene siRNA on the expression and growth of LA-N-5 cells. Methods LA-N-5 cells were transfected with MYCN siRNA marked by folate receptor-targeted liposome-induced Cy3. The transfected siRNA was expressed as fluorescence intensity per μg protein. Changes in MYCN mRNA before and after transfection were detected by real-time quantitative PCR (RT-PCR). Inhibition of LA-N-5 cell growth was assayed by MTT. Results Perμg protein contained (1 808.5 ± 140.2)pg siRNA in LA-N-5 ceils transfected with 100nmol/L MYCN siRNA. The inhibion rate of MYCN mRNA expression and LA-N-5 cell growth was 79.2% and 66.2%, respectively, 72 hours after transfection. Conclusion Folate receptor-targeted liposome-induced MYCN siRNA can significantly inhibit the expression of MYCN mRNA in LA-N-5 cells and the growth of LA-N-5 cells in vitro,which may open up a new way to the gene and target therapy for neuroblastoma.
出处
《军医进修学院学报》
CAS
2010年第2期161-163,共3页
Academic Journal of Pla Postgraduate Medical School
基金
国家自然科学基金项目( 30471802)~~