ABSTRACT
OBJECTIVE To prep are folate-targeted miR- 221 antisense oligonucleotide （anti-miR-221）delivery system ，and to preliminarily evaluate its in vitro anti-cancer effect on hepatocellular carcinoma. METHODS Folate-targeted anti-miR- 221 liposomes（FRL）were prepared by thin-film dispersion method ；the particle size ，Zeta potential and encapsulation efficiency were determined. The delivery efficiency of folate-targeted anionic liposome in human hepatoma HepG 2 cells was determined by in vitro cellular uptake experiment using calcein as the model drug. Flow cytometry was used to detect the effects of FRL on the apoptosis and cell cycle of HepG 2 cells. RESULTS The particle size of prepared FRL was （172.70±3.76）nm，Zeta potential was （－1.16± 0.15）mV and encapsulation efficiency was （83.53±1.85）％. In vitro cellular uptake experiments showed that folate-targeted anionic liposome successfully delivered calcein to HepG 2 cells，and the delivery efficiency in targeted group was higher than that of non-targeted liposome group （P＜0.01）. Apoptosis experiment results showed that the apoptotic rate of HepG 2 cells treated with FRL was significantly higher than that of non-targeted liposome （P＜0.01）. In cell cycle experiment ，FRL could shorten the S phase fraction of HepG 2 cells and induced arrest in the G 0/G1 and G 2/M phases. CONCLUSIONS FRL can encapsulate anti-miR-221 well and deliver it to hepatocellular carcinoma HepG 2 cells successfully ，and has a good in vitro anti-hepatoma effect in inducing apoptosis and cell cycle regulation.
ABSTRACT
OBJECTIVE: To establish effective and reliable methods for the determination of adenovirus and carmustine in anionic liposomes-mediated co-delivery system. METHODS: The complexes of co-delivery system were prepared by calcium-induced phase changing method. For carrying out the quantitative-PCR (Q-PCR) detection, the sequences of primer and probe were designed accoding to the hexon gene sequences of Ad5, and then the reaction system of Q-PCR was optimized to detect the loading rate of Ad5. The HPLC condition was also optimized to determine the drug loading of carmustine in the co-delivery system. RESULTS: In the co-delivery system, the loading rate of adenovirus detected by Q-PCR method was (23.2±1.8)% and that of carmustine was (55±2.8)%. CONCLUSION Both of Q-PCR and HPLC methods have been successfully used for the quantification of adenovirus and carmustine in anionic liposomes-mediated co-delivery system. These methods are simple, reliable and accurate, which can be used in other similar experiments.