PEG化壳聚糖纳米粒介导的hTERT反义寡核苷酸对HepG2细胞的抑制作用

Inhibition of HepG2 cells by PEG-Chitoson nanoparticles loaded with antisense oligodeoxynucleotide of hTERT mRNA

目的以PEG化壳聚糖(PEG-Chitoson)纳米粒作为端粒酶逆转录酶反义寡核苷酸的载体转染HepG2细胞,研究其对HepG2细胞的增殖影响;方法采用MTT法检测PEG-CSNPs的细胞毒性和细胞增殖情况;以脂质体转染试剂作为对照,倒置荧光显微镜和流式细胞仪(FCM)检测FITC标记的ASODN转染细胞后胞内荧光强度,转染效率和ASODN各组转染细胞后细胞周期的变化。结果NPs质量浓度超过3.0μg/mL时细胞毒性较大。转染24h后,ASODN纳米粒组和ASODN脂质体组细胞内荧光明显增强,且ASODN纳米粒组的转染效率要高于ASODN脂质体组。与空白对照组相比,ASODN处理后HepG2细胞生长受到明显抑制(P<0.01),且抑制率随时间延长而增高,72h时达到最高值;细胞周期改变,细胞被阻滞于G1期,S期细胞明显减少(P<0.01);结论PEG-CSNPs介导的hTERT ASODN能有效抑制HepG2细胞增殖、改变细胞周期,对该细胞生长有明显抑制作用。

[Objiective] To investigate the effect of nanoparticles loaded with antisense oligodeoxynucleotide （ASODN） of hTERT mRNA on HepG2 ceils. [Metheods] The eytotoxieity of PEG-CS NPs and proliferation of HepG2 eeils were deteeted by MTT assay. Intraeeilular fluoreseenee intensity, transfeet efficieney and eel/eyeles after transleering ASODN were determined by Inversion fluorescence microscope and flow eytometry （FCM）, and the hpofeetamine acted as the control in this study. [Results] The eytotoxieity increased evidently with the increasing concentration of N-Ps over 3.0 μg/mL The intraeeilular fluorescence in ASODN-NPs group and ASODN-lipofeetamine group were obviously strong after transfeetion for 24 h, and the transfeet efficieney of ASODN-NPs group was higher than that of ASODN-Lipofeetamine group. Compared with the blank group, the inhibitory rate of HepG2 ceils enhanced obviously （P〈0.01） and reached peak at 72 h after treated with ASODN; the ceil cycle was blocked in G1 phase, and the ceil number in S phase decreased obviously （P 〈0.01）. [Conclusion] hTERT ASODN transfect by PEG-CS NPs can inhibit the proliferation and ceil viability of HepG2 ceils effectively and cause the change of ceil eyele.