摘要
目的:探讨中药、化疗药物以及抗血管生成药物联合应用对人肝癌SMMC-7721细胞的协同增效作用及其相关机制.方法:采用荧光显微镜、原位末端标记技术(TUNEL)观察两药组(复方苦参联合奥沙利铂)及三药组[复方苦参联合奥沙利铂和重组人血管内皮抑素(恩度)]对SMMC-7721细胞的诱导凋亡作用;应用免疫组织化学法观察药物作用后对凋亡相关基因Survivin、Bcl-2和caspase-3蛋白表达的影响.结果:荧光显微镜观察发现,复方苦参注射液(50mL/L)和奥沙利铂(1mg/L)两药组以及同时联合恩度(12.5mg/L)的三药组作用于SMMC-7721细胞后,凋亡指数(AI)分别为44.33%和46.33%,两组间未见明显差异(P>0.05);TUNEL检测结果亦显示,两药组与三药组比较,AI分别为36.3%和34.2%,无明显差异(P>0.05);免疫组织化学法观察显示,两药组与三药组均可使Survivin、Bcl-2蛋白表达减弱,caspase-3蛋白表达增强(1.426±0.007,1.442±0.011vs2.485±0.013;1.308±0.012,1.298±0.008vs2.195±0.021;2.503±0.017,2.526±0.013vs1.536±0.007,均P<0.01),但两组之间亦无明显差异.结论:复方苦参注射液、奥沙利铂两药联合以及再与恩度的三药联合均能诱导人肝癌SMMC-7721细胞的凋亡,但三药联合并未能增加诱导凋亡的作用;其诱导凋亡的机制可能与调节细胞Survivin、Bcl-2、caspase-3蛋白的表达有关.
AIM：To investigate the possible synergistic effects of Fufangkushen Injection,oxaliplatin and recombinant human endostatin（Endostar）on apoptosis in human hepatoma cell line SMMC-7721.METHODS：Fluorescence microscopy and TUNEL assay were employed to evaluate apoptosis of SMMC-7721 cells.Immunohistochemistry was used to detect the expression of apoptosis-associated proteins survivin,Bcl-2 and caspase-3 in SMMC-7721 cells.RESULTS：Fluorescence microscopy analysis showed that the apoptosis index（AI）of cells treated with Fufangkushen Injection and oxaliplatin and those treated with Fufangkushen Injection,oxaliplatin and Endostar was 44.33% and 46.33%,respectively,with no significant difference between the two groups（P 0.05）.TUNEL analysis showed that the AI of the above two groups of cells were 36.3% and 34.2%,respectively,with no significant difference between the two groups（P 0.05）.Immunohistochemistry analysis demonstrated that,although the expression of survivin and Bcl-2 decreased and the expression of caspase-3 increased in the above two groups of cells（1.426 ± 0.007,1.442 ± 0.011 vs 2.485 ± 0.013;1.308 ± 0.012,1.298 ± 0.008 vs 2.195 ± 0.021;2.503 ± 0.017,2.526 ± 0.013 vs 1.536 ± 0.007,all P 0.01）,there was no significant difference in the expression levels of these proteins between the two groups.CONCLUSION：Combined Fufangkushen Injection,oxaliplatin and Endostar have comparable apoptosis-inducing ability to combined Fufangkushen Injection and oxaliplatin in SMMC-7721 cells.The above drugs induce SMMC-7721 cell apoptosis possibly by altering the expression of survivin,Bcl-2 and caspase-3.
出处
《世界华人消化杂志》
CAS
北大核心
2010年第32期3396-3401,共6页
World Chinese Journal of Digestology
基金
"十一五"军队中医药重大临床攻关基金资助项目
No.2006051005~~
