论文部分内容阅读
目的:研究叶酸受体介导下的壳聚糖-pGPU6/GFP/Neo纳米粒在叶酸受体表达程度不同的肿瘤细胞中的靶向转染特点。方法:制备叶酸偶联壳聚糖pGPU6/GFP/Neo纳米粒(folate-chitosan-pGPU6/GFP/Neo,FA-CS-DNAnano)和壳聚糖pGPU6/GFP/Neo纳米粒(chitosan-pGPU6/GFP/Neo,CS-DNAnano),红外光谱扫描鉴定其合成,透射电镜观测纳米粒的形态特征及直径大小。纳米粒转染叶酸受体表达阳性的人类卵巢癌细胞系SKOV3、乳腺癌细胞系MCF-7和宫颈癌细胞系HeLa,流式细胞术检测细胞转染效率,MTT法检测纳米粒的细胞毒性。结果:成功制备FA-CS-DNAnano和CS-DNAnano,纳米粒接近球型、表面光滑、结构均匀;FA-CS-DNAnano直径为(78.1±0.3)nm,CS-DNAnano直径为(138.4±0.7)nm。FA-CS-DNAnano在SKOV3和MCF-7细胞的转染效率明显高于CS-DNAnano[(24.3±0.7)%vs(0.7±0.1)%,(16.8±1.2)%vs(0.3±0.1)%;均P<0.01],而在HeLa细胞中两者转染效率无明显差异(P>0.05)。FA-CS-DNAnano转染前后MCF-7、SKOV3和HeLa细胞的活力分别为(87.9±2.4)%、(91.4±1.0)%、(97.4±1.1)%和(63.0±2.5)%、(90.6±1.3)%、(99.3±1.6)%。结论:对于叶酸受体强阳性表达的SKOV3和MCF-7肿瘤细胞,叶酸偶联壳聚糖是良好的靶向基因转运载体。
OBJECTIVE: To study the targeted transfection of chitosan-pGPU6 / GFP / Neo nanoparticles mediated by folic acid receptor in tumor cells with different levels of folate receptor expression. METHODS: The folate-coupled chitosan-pGPU6 / GFP / Neo nanoparticles (chitosan-pGPU6 / GFP / Neo) and chitosan-pGPU6 / GFP / Neo, CS-DNAnano) were synthesized and characterized by FTIR. The morphology and diameter of the nanoparticles were observed by transmission electron microscopy. Nanoparticles were transfected into human ovarian cancer cell line SKOV3, breast cancer cell line MCF-7 and cervical cancer cell line HeLa. The cell transfection efficiency was detected by flow cytometry. The cytotoxicity of nanoparticles was detected by MTT assay. Results: FA-CS-DNAnano and CS-DNAnano were successfully prepared. The diameter of the FA-CS-DNAnano was (78.1 ± 0.3) nm and the diameter of the CS-DNAnano was (138.4 ± 0.7) nm. The transfection efficiency of FA-CS-DNAnano in SKOV3 and MCF-7 cells was significantly higher than that of CS-DNAnano [(24.3 ± 0.7)% vs (0.7 ± 0.1)%, (16.8 ± 1.2)% vs (0.3 ± 0.1)% ; All P <0.01], while there was no significant difference in transfection efficiency between HeLa cells (P> 0.05). The viability of MCF-7, SKOV3 and HeLa cells before and after FA-CS-DNAnano transfection were (87.9 ± 2.4)%, (91.4 ± 1.0)%, (97.4 ± 1.1)% and (63.0 ± 2.5)%, ± 1.3)%, (99.3 ± 1.6)%. CONCLUSION: Folate-conjugated chitosan is a good target gene delivery vector for SKOV3 and MCF-7 tumor cells with strong positive folate receptor expression.