包裹反义寡核苷酸的BCA纳米粒抑制C6脑胶质瘤细胞生长的实验研究

Growth suppressive effect of encapsulating antisense oligodeoxynucleotides in abutylcyanoacrylate nanoparticles on C6 glioma cells

目的优化制备包裹反义寡核苷酸（ASODN）的α-氰基丙烯酸正丁酯（BCA）纳米粒，观察其对C6脑胶质瘤细胞的生长抑制作用。方法以BCA为载体材料，采用界面聚合法制备包裹ASODN的纳米粒（ASODNinNP）．并将其转染至C6脑胶质瘤细胞；倒置显微镜下观察游离ASODN组、ASODNinNP组、吸附ASODN纳米粒组和空白纳米粒组转染C6脑胶质瘤细胞后的细胞生长状态．流式细胞仪（FCM）检测细胞周期变化，采用CCK．8法测定ASODNinNP对细胞毒性和细胞增殖的影响。结果与空白对照组相比．除空白纳米粒组外．其他各组转染后的C6脑胶质瘤细胞形态均发生改变，细胞失去原有的贴壁特性，生长密度降低，生长状态变差，其中以ASODNinNP组最为明显．呈时间依赖性；各组细胞周期均发生变化，表现为G1期细胞比例明显增高，S期细胞比例减少，其中ASODNinNP组最为显著（P〈0．05）；除空白纳米粒组外，各纳米粒组对细胞增殖的抑制效应随ASODN相对终浓度的增加而增加，呈现浓度依赖性特征，其中ASODNinNP组抑制效应显著优于其他各组（P〈0．05）。结论ASODNinNP转染C6脑胶质瘤细胞后能有效抑制细胞增殖及改变细胞周期，对胶质瘤细胞生长有明显抑制作用。

Objective To optimize the preparation of nanoparticles （NP） encapsulating antisense oligodeoxynucleotides （ASODN） and investigate the effects on inhibition of C6 glioma cells. Methods ASODN coated in NP were prepared by interfacial polymerization of butylcyanoacrylate （BCA）. Inverted microscope was used to observe the viability of C6 cells transfected by free ASODN, ASODN in NP, ASODN-NP （ASODN sticking to NP） and BCA-NP, respectively. Cell cycle of C6 cells was studied by flow cytometry （FCM）, and CCK-8 assay was performed to examine the cytotoxicity and proliferation of C6 cells. Results Compared with the control group, all groups, except BCA-NP group, after transfection with NPs appeared cell morphological changes; C6 cells were detached from the matrix, the cell density was reduced and the cell viability was poor; ASODN in NP group was most significant in a time-dependent manner. The cell cycle in ASODN-in-NP group varied obviously compared with the BCA-NP group, and the number of the cells in the G0/G1 phase was increased and the cell number in S phase was decreased significantly （P〈0.05）. The results of CCK-8 assay showed that all groups, but BCA-NP group, produced the inhibition of the cell proliferation to different degrees, and the inhibitory effect was increased with the final concentration increment, especially remarkably in ASODN-in-NP group （P〈0.05）. Conclusion ASODN in NP can inhibit the proliferation and cause cell cycle changes of C6 cells effectively after transfected with ASODN in NP, exerting significantly growth inhibitory effect on C6 glioma cells.