白藜芦醇对肺腺癌A549细胞裸鼠移植瘤的抑制作用

Inhibitory effect of resveratrol on the growth of A549 cell line xenograft in nude mice

目的观察白藜芦醇（Res）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32只荷瘤鼠随机分成对照组和不同浓度Res（10、20、40mg／kg）干预组，连续给药2周，动态观察各组荷瘤鼠体质量和皮下移植瘤体积。2周后处死荷瘤鼠，实时定量聚合酶链反应（Real—timePCR）、Westernblot分别检测皮下移植瘤组织中活化半胱氨酰天冬氨酸特异性蛋白酶（Cleaved—Caspase）-3、生存素（Survivin）的mRNA和蛋白表达水平。同时，体外培养A549细胞，细胞计数试剂盒（CCK-8）法和膜联蛋白V（AnnexinV）一异硫氰酸荧光素（FITC）／碘化丙锭（PI）标记法检测不同浓度Res（10、20、40mol／L）对细胞增殖和凋亡的影响。结果与对照组比较，不同浓度Res对荷瘤鼠体质量均无明显影响（P〉0．05），但对皮下移植瘤体积呈剂量依赖性抑制作用（P〈0．05）。40rag／kgRes干预2周的荷瘤鼠皮下移植瘤体积为对照组荷瘤鼠皮下移植瘤体积的36．1％。Real-timePCR结果表明，与对照组比较，40mg／kgRes显著上调移植瘤组织Cleaved—Caspase-3mRNA表达（1．98±0．46比1．04±0．3，P〈0．05），下调SurvivinmRNA表达（1．15±0．54比2．38±0．67，P〈0．05）。Westernblot结果表明，与对照组比较，40mg／kgRes上调Cleaved—Caspase-3蛋白表达（0．69±0．11比0．04±0．01，P〈0．05），下调Survivin蛋白表达（0．12±0．02比0．94±0．08，P〈0．05）。体外实验证实，40mol／LRes抑制A549细胞增殖，抑制率为（40．69±10．32）％，同时诱导细胞凋亡，凋亡率为（8．85±2．07）％。结论白藜芦醇对肺腺癌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Cleaved—Caspase-3表达，下调Survivin表达抑制细胞增殖，诱导细胞凋亡。

Objective To investigate the effect of Resveratrol （Res） on the growth of A549 cell line xenograft in nude mice and its potential mechanism. Methods The human lung adenoearcinoma xen- ografl mode in nude mice was established with A549 cell. Thirty - two BALB/c - nu mice with lung cancer xenograft were randomly divided into control group and 10, 20, 40 mg/kg Res groups. Different treatments were served for two weeks. The bodyweight of the mice and growth curves of tumor volume were measured every three clays. After two weeks, all mice were sacrificed and the expression of Cleaved - Caspase - 3, Survivin mRNA and protein in subcutaneous tumors were determined by real - time quantitative polymerase chain reaction （ Real - time PCR） and Western blotting. In vitro A549 cell inhibition effects of resveratrol were measured by cell counting kit -8 （CCK -8 ） assay. And apoptosis rates were measured by flow cy- tometry after Annexin V - fluorescein isothiocyanate （FITC）/propidium Iodide （PI） staining. Results In vivo Res effectively inhibited the growth of lung cancer in a dose dependent manner in nude mice （ P 〈 0, 05 ）. No evident differences in weight among different groups were observed （ P 〉 0. 05 ）. The volume of subcutaneous transplantation tumor in 40 mg/kg Res group had 36. 1 percent of that of control group after 2 weeks. Real - time PCR result showed that 40 mg/kg Res up - regulated Cleaved - Caspase - 3 （ 1.98 ±0.46vs. 1.04±.3, P〈0.05） and down-regulated Survivin （1.15±.54 vs. 2. 38 ±0.67, P〈 0. 05 ） mRNA expression. Western blotting result showed that 40 mg/kg Res up - regulated Cleaved - Caspase - 3 （0. 69 ±0. 11 vs. 0. 04 ± 0. 01, P 〈 0. 05 ） and down - regulated Survivin （0. 12 ±0. 02 vs. 0. 94 ± 0. 08, P 〈 0. 05 ） protein expression. In vitro 40 mg/kg Res inhibited the proliferation of A549 cell with the inhibition rate （40. 69 ± 10. 32） % , and induced the apoptosis of A549 cell with the apoptosis rate （ 8.85 ± 2. 07 ） %. Conclusion Res could inhibit the growth of A549 cell line xenograft tumor in nude mice. The potential mechanism may inhibit the proliferation of and induced the apoptosis of A549 cell by increasing the expression of Cleaved - Caspase - 3 and reducing the expression of Survivin.