9-硝基喜树碱及其脂质体对HepG2、L02细胞周期和凋亡作用的研究  

Modulating effects of 9-nitrocamptothecin and its liposomes to induce apoptosis and inhibit cell cycle in HepG2 and L02 cell lines

作　　者：郑顺贞[1] 卢进利[1] 陈智慧[1] 何斌[2] 邹声泉[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院普外科,武汉430030 [2]四川大学生物材料工程研究中心

出　　处：《中华肝胆外科杂志》2011年第5期416-419,共4页Chinese Journal of Hepatobiliary Surgery

基　　金：国家863计划资助（编号：2007AA021801）

摘　　要：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，Western Blot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0．1umol／L时24h后HepG2细胞完全阻滞于S期；0．1μmol／L孵育72h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、Caspase-3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。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 Ceils 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/TAAD. 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 lipo- somes, 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 0.1μmol/L after incubation for 72 h. Apop tosis induction effect also showed a time-dependent and dose-dependent manner. Western Blot results showed the expression of Bax and Caspase-3 were upreguiated 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 lipo somes 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.

关 键 词：肝癌 9-硝基喜树碱 脂质体 细胞周期 凋亡 

分 类 号：R285[医药卫生—中药学]