银杏叶提取物对泪腺腺样囊性癌ACC-2细胞增殖及Survivin,TIP30基因表达的影响  

Influence of Ginkgo biloba extract on proliferation of ACC-2 cell,Survivin and TIP30 gene expression in adenoid cystic carcinoma of lacrimal gland

作　　者：牛坡[1] 赵新霞[2] 燕飞[3] 周雍明[4] 简鹏 

机构地区：[1]河南省人民医院肿瘤放射科,河南郑洲450003 [2]河南省食品药品检验所,河南郑洲450003 [3]郑州人民医院中医科,河南郑州450012 [4]中国中医科学院广安门中医院肿瘤科,北京100053 [5]河南推拿职业学院,河南洛阳471000

出　　处：《中国中药杂志》2014年第24期4860-4864,共5页China Journal of Chinese Materia Medica

摘　　要：探讨银杏叶提取物(EGB)对泪腺腺样囊性癌ACC-2细胞增殖、凋亡的影响,并从基因和蛋白水平上分析EGB对Survivin和TIP30基因表达的影响。不同浓度EGB处理体外培养的人泪腺腺样囊性癌ACC-2细胞,应用MTT法检测细胞增殖;Annexin V/PI双染色流式细胞仪检测细胞凋亡和细胞周期;RT-PCR和Western blotting分析Survivin,TIP30基因和蛋白的表达。结果发现EGB对ACC-2细胞的体外增殖具有抑制作用,量效关系显著,与对照组比较有统计学差异(P<0.01),半数抑制浓度(IC50)为88 mg·L-1。经流式细胞仪检测表明,EGB能使ACC-2细胞G0/G1期逐渐增加,G2/M期和S期逐渐减少,并且随着剂量的增加,ACC-2细胞凋亡率明显增加(P<0.05或P<0.01)。RT-PCR与Western杂交结果一致表明随着EGB浓度的升高,Survivin基因表达明显减少(P<0.01),而TIP30基因表达则显著提高(P<0.01)。因此,EGB能有效抑制人泪腺腺样囊性癌ACC-2细胞Survivin基因表达,并促进TIP30的表达,诱导ACC-2细胞凋亡,从而抑制肿瘤细胞增殖。该研究为中药成分在肿瘤治疗中的应用提供了一定的理论和实验依据。Exploring the influence of extract of Ginkgo biloba （EGB） on the proliferation, apoptosis of ACC-2 cell in lacrimal adenoid cystic carcinoma and analyzing the influence of EGB on the gene expression of Survivin and TIP30 based on the levels of the gene and protein. ACC-2 cell in human with ACC of lacrimal gland disposed by EGB of different concentration was in vitro cultured. MTI＇ method was used for cell proliferation detection. Annexin V/PI double-staining flow cytometer was used to detect cell apoptosis and cell cycle. Survivin and TIP30 gene expression together with protein expression were analyzed by RT-PCR and Western blotting. And it is indicated that EGB has inhibitory effect on the proliferation of ACC-2 cell in vitro. Furthermore, the dose-effect relationship was significant. Compared with the control group, it had statistical difference （P 〈 0.01 ）. The inhibitory concentration 50% （ IC50） is 88 mg ·L^-1. By flow cytometer examination, it was indicated that EGB can gradually increase ACC-2 cell in G0-G1 stage and decrease it in G2-M and S stage. With the increase of dose, the apoptosis rate of ACC-2 cell obviously increased （P 〈 0.05 or P 〈 0.01 ）. Both of the expression results of RT-PCR and Western hybrid proteins have showed that the concentration of EGB increased, it could be seen a significant decrease in Survivin gene expression （P 〈 0.01）. Meanwhile, the TIP30 gene expression got a significant increase. Therefore, EGB can effectively inhibit ACC-2 cell Survivin gene expression in human with adenoid cysistic carcinoma of larcrimal gland as well as promoting TIP30 gene expression, inducing the ACC-2 cell apoptosis and inhibiting tumor cell proliferation, which provided a certain theoretical and experimental basis for the application of Chinese herbal medicinal ingredient in the treatment of tumors.

关 键 词：银杏叶提取物 ACC-2细胞 Survivin基因 TIP30基因 

分 类 号：R285[医药卫生—中药学]