反义ClC-3寡核苷酸对PC12细胞增殖的影响

Effect of ClC-3 antisense oligonucleotide on proliferation of PC12 cells

目的:研究反义ClC-3寡核苷酸对PC12细胞增殖的影响。方法:PC12细胞用Lipofectamine2000介导分别转染6.25,12.5,25.0,50.0,100.0及200.0mg/L的CIC-3反义寡核苷酸,50mg/LCIC-3正义寡核苷酸,50mg/LClC-3随义寡核苷酸,以转染10mg/LLipofectamine2000和未转染细胞作对照。利用MTT以及[3H]-胸腺嘧啶掺入实验检测细胞存活率及DNA合成情况,流式细胞术检测细胞周期。结果:与未转染细胞组比较,反义ClC-3寡核苷酸可以降低PC12细胞存活率,并抑制[3H]-胸腺嘧啶掺入到DNA,使G0/G1期细胞增多,S期细胞减少(P均<0.05)。而转染脂质体、正义及随义ClC-3寡核苷酸对PC12细胞的细胞存活率、DNA合成及细胞周期均无影响(P>0.05)。结论:反义ClC-3寡核苷酸通过使PC12细胞周期被阻滞在G0/G1期而抑制细胞的增殖。

Aim： To investigate the effect of CIC-3 antisense oligonucleotide on the proliferation of PCI2 cells. Meth- ods： PC 12 cells were grouped to transfect 6.25,12.5,25.0,50.0,100.0 and 200.0 mg/L CIC-3 antisense oligonucleotide, 50 mg/L CIC-3 sense oligonucleotide,50 mg/L CIC-3 missense oligonucleotide, respectively, cells without any transfection and transfected 10 mg/L Lipofeetamine^2000were the control, MTT and [ 3HI-TdR incorporation assay were used to measure the viability and I）NA synthesize, and cell cycle was studied with flow cytometry analysis. Results： Compared with the con- trol, CIC-3 antisense oligonueleotide could decreased the viability of PCI2 cells, and the [3H]-TdR incorporation into DNA, and increased the numbers of the cells in G1 stage but decreased that in S stage（ P 〈 0.05）. Whereas lipofectamine, missense or sense oligonucleotide transfection had no effects on the proliferation of PCI2 cells（ P 〉 0.05 ）. Conclusion： CIC-3 antisense oligonucleotide could arrest PCI2 cells in G0/G1 stage which results in the inhibition of PCI2 cells proliferation.