CAJ | 학술논문

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(ASODN in NP),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、ASODN in NP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ASODN in NP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ASODN in NP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ASODN in NP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ASODN in NP组抑制效应显著优于其他各组(P<0.05).结论 ASODN in NP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.
목적 우화제비포과반의과핵감산(ASODN)적α-청기병희산정정지(BCA)납미립,관찰기C6뇌효질류세포적생장억제작용.방법 이BCA위재체재료,채용계면취합법제비포과ASODN적납미립(ASODN in NP),병장기전염지C6뇌효질류세포;도치현미경하관찰유리ASODN조、ASODN in NP조、흡부ASODN납미립조화공백납미립조전염C6뇌효질류세포후적세포생장상태,류식세포의(FCM)검측세포주기변화,채용CCK-8법측정ASODN in NP대세포독성화세포증식적영향.결과 여공백대조조상비,제공백납미립조외,기타각조전염후적C6뇌효질류세포형태균발생개변,세포실거원유적첩벽특성,생장밀도강저.생장상태변차,기중이ASODN in NP조최위명현,정시간의뢰성;각조세포주기균발생변화,표현위G1기세포비례명현증고,S기세포비례감소,기중ASODN in NP조최위현저(P<0.05);제공백납미립조외,각납미립조대세포증식적억제효응수ASODN상대종농도적증가이증가,정현농도의뢰성특정,기중ASODN in NP조억제효응현저우우기타각조(P<0.05).결론 ASODN in NP전염C6뇌효질류세포후능유효억제세포증식급개변세포주기,대효질류세포생장유명현억제작용.
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 NT were prepared by interfacial polymerization of butyleyanoacrylate (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 GO/GI 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.