CAJ | 학술논문

目的研究9-硝基喜树碱及其脂质体对人肝癌细胞株HepG2和人正常肝细胞L02细胞周期、凋亡的影响及其可能的作用机制.方法 HepG2、L02细胞用含不同浓度9-硝基喜树碱及其脂质体的培养液孵育后,利用MTT法测定细胞活性,流式细胞术检测细胞周期及细胞凋亡,WesternBlot验证周期相关蛋白和凋亡相关蛋白的表达变化.结果 9-硝基喜树碱及其脂质体对两种细胞生长呈时间和剂量依赖性抑制.药物处理后S期和G2/M期细胞明显增多,浓度大于0.1 μmol/L时24 h后HepG2细胞完全阻滞于S期;0.1 μmol/L孵育72 h后,超过95%HepG2细胞阻滞于G2/M期.药物对细胞凋亡的诱导作用也呈明显的剂量和时间依赖关系.Western Blot显示:Bax、Caspase3表达增高,Cyclin A、Cdk2、Cyclin E、Bcl-2表达减低.与L02相比,HepG2细胞对药物更加敏感.结论 9-硝基喜树碱及其脂质体可以通过调控细胞周期和诱导凋亡有效抑制细胞生长,其对肿瘤细胞的抑制作用强于正常肝细胞.9-硝基喜树碱脂质体体外抗肿瘤效果略强于9-硝基喜树碱单体.
목적 연구9-초기희수감급기지질체대인간암세포주HepG2화인정상간세포L02세포주기、조망적영향급기가능적작용궤제.방법 HepG2、L02세포용함불동농도9-초기희수감급기지질체적배양액부육후,이용MTT법측정세포활성,류식세포술검측세포주기급세포조망,WesternBlot험증주기상관단백화조망상관단백적표체변화.결과 9-초기희수감급기지질체대량충세포생장정시간화제량의뢰성억제.약물처리후S기화G2/M기세포명현증다,농도대우0.1 μmol/L시24 h후HepG2세포완전조체우S기;0.1 μmol/L부육72 h후,초과95%HepG2세포조체우G2/M기.약물대세포조망적유도작용야정명현적제량화시간의뢰관계.Western Blot현시:Bax、Caspase3표체증고,Cyclin A、Cdk2、Cyclin E、Bcl-2표체감저.여L02상비,HepG2세포대약물경가민감.결론 9-초기희수감급기지질체가이통과조공세포주기화유도조망유효억제세포생장,기대종류세포적억제작용강우정상간세포.9-초기희수감지질체체외항종류효과략강우9-초기희수감단체.
Objective To investigate the modulating effects and explore their mechanism of 9-nitrocamptothecin and its liposomes to induce apoptosis and inhibit cell cycle in HepG2 and L02 cell lines. Methods Cells were incubated with 9-nitrocamptothecin(9NC) or with 9-nitrocamptothecin liposomes for 24 h, 48 h and 72 h, then, the cell viability was measured via MTT assay; cell cycle and apoptosis was evaluated by flow cytometry after stained by PI and Annexin V-PE/7AAD. Additional, Western Blot was used to evaluate the expression of cell cycle and apoptosis related protein. Results Both cells viability were apparently inhibited by the 9-nitrocamptothecin and 9-nitrocamptothecin liposomes, the inhibitory effect showed a time-dependent and dose-dependent manner. Both S and G2/M phases arrest were observed after incubated with drugs. HepG2 cell was completely arrested in S phase when 9NC concentration over than 0. 1 μmol/L after incubation for 24 h, while more than 95% cells arrested in G2/M phase when 9NC concentration is 0. 1 μmol/L after incubation for 72 h. Apoptosis induction effect also showed a time-dependent and dose-dependent manner. Western Blot results showed the expression of Bax and Caspase-3 were upregulated while Cyclin A, Cdk2, Cyclin E and Bcl-2 were downregulated. More importantly, the compounds were more cytotoxic to the cancer cell lines than to the normal liver cell. Conclusions 9-nitrocamptothecin and 9-nitrocamptothecin liposomes can potently inhibit cell growth via regulation of cell cycle and induction of apoptosis, and this effect was preferentially in cancer cell. Inhibitory of 9-nitrocamptothecin liposomes was slightly better than the 9-nitrocamptothecin.