目的 考察阳离子脂质体(CL)为载体的反义寡核苷酸(ASODN)序列对HeLa细胞的抑制作用。方法 选择以人端粒酶模板RNA(hTR)和人逆转录酶催化亚基(hTERT)为靶点的两种ASODN,用3β[N-(N′,N′-二甲氨基乙基)氨甲酰基]-胆固醇(DC-Chol)和二油酰基磷脂酰乙醇胺(DOPE)制备CL,考察其形态和粒径分布,并用其介导反义寡核苷酸转染HeLa细胞,通过噻唑蓝比色法(MTT)研究对细胞的抑制作用。结果 CL外观呈圆形,粒径均匀,在体外能够有效转染反义hTR和反义hTERT寡核苷酸序列(anti-hTR,anti-hTERT)。CL/ASODN处理后的HeLa细胞,72h后的存活率为(19.82±12.19)%(anti-hTR,2.0μmol·L^-1)和(16.59±4.10)%(anti-hTERT,2.0μmol·L^-1),而相同浓度游离的ASODN没有发现抑制作用。CL/ASODN复合物能够在120h内持续抑制HeLa细胞的生长。结论 CL能够有效提高ASODN对HeLa细胞的抑制作用,作为端粒酶抑制剂的新型非病毒载体,具有良好的应用前景。
OBJECTIVE To investigate the growth inhibition of antisense oligonucleotides ( ASODN ) mediated by cationic liposomes(CL) on HeLa ceils. METHODS Two ASODNs targeting at human telomerase template RNA (hTR) and human telomerase catalytic subunit (hTERT) ,respectively ,were designed ( anti-hTR ,anti-hTERT). CL was prepared using 3β[ N- ( N' ,N'-dimethylaminoethan) -carbamoyl ] cholesterol (DC-Chol) and 1,2-Dioleoyl-sn-glycero-3-phosphoethanolamine ( DOPE ). The properties of CL, such as the morphology and average size,were detected. ASODN loaded CL were transfected to HeLa ceils, and the growth inhibition was identified by MTT assay. RESULTS All CL were spherical with narrow size distribution. ASODN was efficiently transfected by CL in vitro. After 72 h,the survival rates of HeLa ceils were only ( 19. 82 ± 12. 19)% (anti-hTR,2. 0 μmol · L^-1 ) and ( 16.59 ±4. 10)% (anti-hTERT,2.0 μmol · L^-1) when being treated with CL/ASODN complex,while no inhibition effect was found in free ASODN (2. 0 μmol·L^-1 ) groups. Growth inhibition in HeLa ceils by CL/ASODN complexes was observed within 120 h. CONCLUSION CL significantly enhance the growth inhibition effect of ASODN on tumor cells, which suggestes that CL can be a promising non-vlral vector used for telomerase inhibitor.