摘要
目的研究体内外紫杉醇处理后，胃癌细胞survivin蛋白表达的改变及同细胞凋亡关系，探讨紫杉醇抗肿瘤机制。方法以不同浓度紫杉醇（0、12、60、300、1500ng／mL）分别处理胃癌BGC-823细胞24h，检测survivin蛋白表达；另以600ng／mL紫杉醇处理该细胞72h，检测处理0、12、24、36、48h时survivin表达及反映细胞凋亡程度的PARP89000片段。以BGC-823细胞构建裸鼠移植瘤模型，予紫杉醇40mg／kg腹腔注射，分别在干预后0、1、3、5、7、14d处死裸鼠，取移植瘤，一半固定行免疫组化检测；一半液氮冻存留行Westernblot检测。结果紫杉醇处理BGC-823细胞24h时，survivin表达升高具有紫杉醇剂量依赖性。600ng／mL紫杉醇处理24h内，survivin表达逐步升高，36h后明显下降，48h时更低；处理36h和48h时出现明显的PARP89000片段。裸鼠移植瘤结果显示，紫杉醇处理后第3天瘤体内坏死区明显增多，第5～7天达高峰，第14天又有缩小；免疫组化结果显示干预前survivin表达强阳性，干预后1d变化不明显，第3天明显减弱、第5～7天时最低，第14天又开始升高。Westernblot也得出同样结果。结论体内外试验结果证实，紫杉醇可能通过抑制肿瘤survivin表达，引起细胞凋亡，从而抑制肿瘤生长。体外干预早期survivin表达升高可能为细胞的一种保护作用。
Objective To detect the variation of survivin expression and cell apoptosis in gastric cancer cell line of BGC-823 after taxol administration, and to invest the antitumor mechanism of taxol. Methods Taxol was administered to gastric cancer cell line of BGC-823 at different concentration for 24 h or at 600 ng/mL for 0, 12, 24, 36, 48, 72 h, and then survivin expression and PARP89000 frag- ment which indicating the extent of apoptosis were detected by western blot method. The relationship between apoptosis and survivin expression was analyzed. Nude mice transplanted tumor models were made with BGC-823 cell line. Total 18 nude mice were classified into 6 groups randomly. Five groups of them were given intraperitoneal injection of taxol at the dose of 40 mg/kg, and the other group used as control and never received any treatment. These nude mice were killed at day0, 1, 3, 5, 7, 14 respectively, and survivin expression in the tumor tissue was detected by both immunohistochemical analysis and western blot. Results Survivin expression increased gradually during 24 h after administration of taxol, and the increase had a positive correlation with taxol＇s concentration. When treatment longer than 24 h, survivin expression dropped at 36 h, and still lower at 48h. High leve; of PARP89000 fragment, which indicating the occurrence and the extent of apoptosis, was detected at 36 h and 48 h after administration of taxol. The nude mice experiments indicated that the necrosis areas in transplanted tumor tissue increased 3 days after administrated of taxol. The necrosis area extended to its maximum at 5 and 7 days after treatment of taxol. Immunohistochemical results showed that survivin expression was strong positive at day 0 and day 1, then dropped down since day 3. At day 5 and 7, survivin expression dropped to the minimum point ,but it recovered to be heavey positive at day 14. The western blot result showed similar output. Conclusions Taxol can inhibit survivin expression, and prevent cell prolifration. This could be an impo
出处
《复旦学报（医学版）》
CAS
CSCD
北大核心
2008年第1期67-71,共5页
Fudan University Journal of Medical Sciences
基金
复旦大学青年基金资助项目(JKF159001)
关键词
胃癌
紫杉醇
生存蛋白
gastric carcinoma
survivin
paclitaxol
