Inhibitory effect of resveratrol on the growth of human colon cancer ls174t cells and its subcutaneously transplanted tumor in nude mice and the mechanism of action
Inhibitory effect of resveratrol on the growth of human colon cancer ls174t cells and its subcutaneously transplanted tumor in nude mice and the mechanism of action
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DOI：
10.3760/cma.j.issn.0253-3766.2009.01.004
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发表时间：
2009-01-01
期刊：
影响因子：
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通讯作者：
Guo Xing-gang
中科院分区：
文献类型：
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作者：
Chen Jie;Dong Xin-shu;Guo Xing-gang
目的探讨白藜芦醇对体外培养的人结肠癌ls 174 t细胞及荷瘤裸鼠结肠癌生长的影响及其机制。方法采用MTT法检测白藜芦醇对ls 174 t细胞生长增殖的抑制作用。透射电镜观察细胞凋亡的形态学变化，流式细胞仪检测细胞凋亡率和细胞周期的变化。RT-PCR检测bcl-2和bax mRNA的表达，Western blot检测bcl-2 bax蛋白的表达。结果MTT法显示白藜芦醇对ls 174 t细胞生长有明显的抑制作用，并呈浓度和时间依赖性。在25、50、100、200和400 μ mol/L浓度范围内，白藜芦醇处理24 h后的抑制率分别为1.0%、9.1%、17.4%、27.8%和66.5%，处理48 h后的抑制率分别为3.6%、13.7%、30.2%，作用72 h后抑制率分别为18.1%、33.0%、48.6%、61.2%和89.4%，差异极显著（P < 0.01）。白藜芦醇处理的ls 174 t细胞超微结构出现典型的凋亡改变。流式细胞仪检测发现白藜芦醇可诱导ls 174细胞凋亡，并使细胞周期阻滞于S期。RT-PCR和Western blot检测显示，不同浓度（25、50、100和200 μ mol/L）的白藜芦醇处理结肠癌细胞后，bcl-2表达水平降低，bax表达水平升高。最高抑制率为47.9%。200 mg/kg和800 mg/kg白藜芦醇治疗组裸鼠皮下移植瘤重分别为4.10 ± 0.18 g和3.05 ± 0.35 g，与对照组比较差异有显著性（P < 0.01）。结论白藜芦醇可通过诱导凋亡抑制ls 174 t细胞的生长，其机制可能与抑制抗凋亡因子bcl-2和增强凋亡因子bax的表达有关。
Objective To explore the effect and mechanism of resveratrol against human colon cancer ls174t cells in vitro and the growth of colon cancer in tumor-bearing nude mice. Methods MTT method was used to test the inhibiting effect of resveratrol on the growth and proliferation of ls174t cells. Transmission electron microscopy was used to observe the morphological changes of cell apoptosis, and FCM assay was performed to measure the changes of cell apoptosis rate and cell cycle. RT-PCR method was used to defect the expression of bcl-2 and bax mRNA, and Western blot was used to detect the expression of bcl-2 bax protein. Results MTT test revealed that resveratrol showed significant inhibiting effect on ls174t cells in a concentration- and time-dependent manner. In the concentration range of 25, 50, 100, 200 and 400 mu mol/L, the inhibition rate after resveratrol treatment for 24 hours was respectively 1.0%, 9.1%, 17.4% 27.8% and 66.5%, while the inhibiton rate after treatment for 48 hours was respectively 3.6%, 13.7%, 30.2%, 58.4% and 86.1% and the inhibition rate after treatment for 72 hours was 18.1%, 33.0%, 48.6%, 61.2% and 89.4%, respectively, showing a very significant difference (p < 0.01). Typical ultrastructural apoptotic changes were observed in resveratrol-treated ls174t cells. It was found through FCM assay that resveratrol caused apoptosis in ls174 cells and blocked the cell cycle at S phase. RT-PCR and Western blot test showed that after the treatment of colon cancer cells with resveratrol at different concentrations (25, 50, 100 and 200 mu mol/L), the expression level of bcl-2 was decreased, while expression level of bax was increased. The highest inhibition rate was 47.9%. In 200 mg/kg and 800 mg/kg resveratrol treatment groups, the weight of subcutaneously transplanted tumors in nude mice was 4.10 +/- 0.18 g and 3.05 +/- 0.35 g, respectively, the difference was significant compared with that of the control group (P < 0.01). Conclusion Resveratrol can inhibit the growth of ls174t cells through apoptosis induction, The mechanism is probably related to inhibition of anti-apoptotic factor bcl-2 and enhancement of expression of apoptotic factor bax.