摘要
目的 观察姜黄素对肺腺癌A549细胞裸鼠移植瘤的干预作用并探讨机制.方法 建立肺腺癌A549细胞裸鼠移植瘤模型,将40只荷瘤鼠随机分成对照组和不同浓度（10、50、100 mg/kg）姜黄素干预组,每隔1d腹腔注射给药1次,共8次,每4d测定各组荷瘤鼠体质量和皮下移植瘤体积.同时体外培养A549细胞,细胞计数试剂盒（CCK-8）和膜联蛋白V（Annexin V）-异硫氰酸荧光素（FITC）/碘化丙锭（PI）标记法分别检测不同浓度姜黄素（10、50、100 mmol/L）对细胞增殖和凋亡的影响.实时定量反转录聚合酶链反应（RT-qPCR）和Western blot分别检测不同处理组细胞B细胞淋巴瘤/白血病-2相关X蛋白（bax）和B细胞淋巴瘤/白血病-2（bcl-2）mRNA和蛋白表达水平.半胱氨酰天冬氨酸特异性蛋白酶（Caspase）-3活性检测试剂盒测定各组细胞Caspase-3活性相对值.结果 与对照组比较,不同浓度姜黄素对荷瘤鼠体质量均无明显影响（P＞0.05）,但10、50、100 mg/kg姜黄素干预荷瘤鼠16d后皮下移植瘤平均体积分别为同期对照组的76.2％、55.7％和38.3％,抑制作用呈浓度依赖性（P＜0.05）.10、50、100 mmol/L姜黄素均能抑制体外培养的A549细胞增殖,同时促进细胞凋亡,100 mmol/L姜黄素干预A549细胞48 h后抑制率为（51.95±13.32）％,凋亡率为（30.89-±6.27）％.RT-qPCR和Western blot结果均表明,100 mmol/L姜黄素显著上调bax表达,下调bcl-2表达,使bax/bcl-2比值增大,且Caspase-3活性相对值也达到峰值（8.39±1.40）,与其他各组比较差异均有统计学意义（P＜0.05）.结论 姜黄素对肺腺癌A549细胞裸鼠移植瘤的生长具有抑制作用,其机制可能是通过上调bax表达,下调bcl-2表达,活化下游Caspase-3信号,从而抑制细胞增殖,诱导细胞凋亡.
Objective To investigate the effect and mechanism of curcumin on the growth of A549 cell line xenograft in nude mice.Methods The human lung adenocarcinoma xenograft model in nude mice was established with A549 cells.Forty BALB/c-nu mice with lung cancer xenograft were randomly divided into control group and 10,50,100 mg/kg curcumin groups.Different treatments were given by peritoneal injection every other day for 8 times.The bodyweight of mice and the volume of tumor were measured every 4 days.In vitro the proliferation of A549 cell was determined by cell counting kit-8 （CCK-8） assay,and the apoptosis rate was measured by flow cytometry after Annexin V-fluoresceine isothiocyanate （FITC）/propidium iodide （PI） staining.The expression of B cell lymphoma/leukemia-2 associated X protein （bax） and B cell lymphoma/lenkemia-2 （bcl-2） mRNA and protein was detected by real-time reverse transcriptase-polymerase chain reaction （RT-qPCR） and Western blotting respectively.And Cysteinyl aspartate-specific protease-3 （Caspase-3） relative activity was measured by Caspase-3 assay kit.Results In vivo 10,50,100 mg/kg of curcumin effectively inhibited the growth of tumor in nude mice （P 〈 0.05）,especially in 100 mg/kg group.No evident differences in weight among different groups were observed （P 〉0.05）.In vitro 10,50,100 mmol/L of curcumin inhibited the proliferation of A549 cells,but induced the apoptosis of cells.The inhibition rate of A549 cells in 100 mmol/L curcumin group after 48 h treatment was （51.95 ± 13.32） %,and the apoptosis rate was （30.89 ± 6.27） %.The results of RT-qPCR and Western blotting showed that 100 mmol/L curcumin significantly up-regulated the bax expression and down-regulated the bcl-2 expression,leading to the maximum ratio of bax/bcl-2.In addition,Caspase-3 relative activity also reached the peak value （8.39 ± 1.40） in 100 mmol/L curcumin group,with significant differences among groups （P 〈 0.05）.Conclusion Curcumin was able to inhibit the growth of A549 cell line xenograft tumor in nude mice.The potential mechanism was that curcumin inhibited the proliferation of and induced the apoptosis of A549 cells by increasing the expression of bax and reducing the expression of bcl-2,leading to the activation of down stream Caspase-3 signaling pathway.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2016年第10期2358-2361,共4页
Chinese Journal of Experimental Surgery
基金
湖北省自然科学基金（2015CFA027）
湖北省卫生计划生育委员会科研基金（WJ2015MA010）
