莪术对SGC7901胃癌细胞COX-1,COX-2,VEGF和PGE_2表达的影响  

Effect of Ezhu on expression of cyclooxygenase, vascular endothelial growth factor and prostaglandin E_2 in human gastric cancer cell line SGC7901

作　　者：沈洪[1] 刘增巍[1] 朱萱萱[2] 张坤[3] 王伟[3] 郭青龙[3] 袁胜涛[3] 

机构地区：[1]江苏省中医院消化科,江苏省南京市210029 [2]江苏省中医院动物实验室,江苏省南京市210029 [3]中国药科大学肿瘤药理实验室,江苏省南京市210009

出　　处：《世界华人消化杂志》2006年第16期1548-1553,共6页World Chinese Journal of Digestology

基　　金：江苏省自然科学基金资助项目;No.BK2001186~~

摘　　要：目的:观察莪术对人胃癌SGC7901细胞 COX-1,COX-2,VEGF和PGE2的影响．方法:MTT法观察莪术对胃癌细胞的抑制作用,绘制其抑制率曲线,选取抑制率为25％时的药物浓度作为加药浓度,用RT-PCR以及Western blot方法检测加药后人胃癌细胞 COX-1,COX-2基因及其蛋白表达的变化．用ELISA方法检测加药后培养基中VEGF和 PGE2的表达情况．结果:莪术对胃癌细胞有一定的抑制作用,且随着浓度的增加,其抑制作用加强;将RT-PCR 产物经电泳分析,证实胃癌细胞中有COX-1、 COX-2基因的表达,对各条带进行灰度分析发现:中西药对COX-1和COX-2均有抑制作用,莪术对COX-2的抑制作用大于塞来昔布;Western blot曝光结果可见,COX-1各组在 70 kDa处可见特异性的蛋白条带,COX-2各组在80 kDa处可见特异性的蛋白条带．对各条带进行灰度分析发现:各组中西药对COX-2 均有明显的抑制作用,而对COX-1则无抑制作用,其中,莪术对COX-2的抑制作用要强于celecoxib;西药celecoxib及莪术组可明显降低人胃癌细胞VEGF的表达,与细胞组相比,其差别有统计学意义(91．0±18．2,127．8 ±12．1 ng／L vs162．0±15．1 ng／L,P<0．01);西药celecoxib组PGE2表达低于细胞组,但其差别无统计学意义,莪术可明显降低人胃癌细胞PGE2的表达,与细胞组相比,其差别有统计学意义(67．5±6．9 ng／L vs 78．7±5．6 ng／L, P<0．01)．结论:莪术可能是通过抑制COX-2及其下游 PGE2表达,使VEGF表达下调而抑制肿瘤．AIM： To investigate the effect of Ezhu on the expression of cyclooxygenase-1 （COX-l）, cyclooxygenase-2（COX-2）, vascular endothelial growth factor （VEGF） and prostaglandin E2 （PGE2） in human gastric cancer cell line SGC7901. METHODS： The MTT method was used to ob-serve the suppression of the human gastric cancer （SGC7901） cells treated with Ezhu. By drawing the suppression rate curves, we selected the appropriate Ezhu concentration at the 25% proliferation suppression rate as experimental concentration. Reverse transcription-polymerase chain reaction （RT-PCR） and Western blot were adopted, respectively, to examine the expression of COX gene and protein. Furthermore, ELISA method was used to detect the variation of the VEGF and PEG2 expression in culture medium. RESULTS： Ezhu definitely inhibited the proliferation of human gastric cancer cells in a concentration dependent manner. Electrophoresis of the RT-PCR products demonstrated the expression of COX-1 and COX-2 gene in the human gastric cancer cells. Gray scale analysis showed that both Ezhu and celecoxib inhib- ited the expression of COX-1 and COX-2 gene, and the inhibitory action of Ezhu was greater than celecoxib. However, Western blot clearly showed that a specific protein strip was observed in COX-1 group at the 70 kDa, while in COX-2 group at the 80 kDa. Gray scale analysis confirmed than both Ezhu and celecoxib had ap- parent suppression on COX-2 protein, but had no effect on COX-1. Moreover, the suppression function of Ezhu surpassed celecoxib. Ezhu and celecoxib obviously degraded the content of VEGF in gastric cancer cells. In comparison with those in the control group, the results are statistically significant （91.0±18.2, 127.8±12.1 ng/L vs 162.0±15.1 ng/L, P〈0.01）. The level of PGE2 in the cells treated with celecoxib was lower than the control group with no statistical significance. Nevertheless, Ezhu decreased the level of PGE2 significantly （67.5±6.9 ng/L vs 78.7±5.6 ng/L, P〈 0.01）. CONCLUSION�

关 键 词：莪术 胃癌 SGC7901细胞 环氧合酶 血管内皮生长因子 前列腺素E2 

分 类 号：R735.2[医药卫生—肿瘤]